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  • 1
    ISSN: 0749-503X
    Keywords: recombinant DNA ; K. lactis genomic library ; pCXJ22 ; arginine biosynthesis ; KlARG8 ; mitochondrial transformation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A recombinant plasmid was isolated from a Kluyveromyces lactis genomic DNA library which complements a Saccharomyces cerevisiae arg8 mutant defective in the gene encoding acetylornithine aminotransferase. The complementation activity was found to reside within a 2.0 kb DNA fragment. Nucleotide sequence analysis revealed an open reading frame able to encode a 423-residue protein sharing 68·1% and 35·0% sequence identities with the products of the ARG8 and argD genes of S. cerevisiae and Escherichia coli. That the cloned gene, KlARG8, is the functional equivalent of S. cerevisiae ARG8 was supported by a gene disruption experiment which showed that K. lactis strains carrying a deleted chromosomal copy of KlARG8 are auxotrophic for arginine. The nucleotide sequence of KlARG8 has been submitted to GenBank under Accession Number U93209.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 11 (1995), S. 1613-1627 
    ISSN: 0749-503X
    Keywords: yeast ; brewing ; Saccharomyces ; genetic-modification ; recombinant DNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Academic studies and traditional breeding of yeasts depend upon their sporulation lifestyle. The strains used have been specially selected to sporulate readily and to mate producing new yeast types. Unfortunately brewing yeast strains do not behave in this way. They sporulate poorly, any spores which are formed are usually non-viable and any haploid strains produced are invariably non-maters. Only in recent years, with the development of recombinant-DNA techniques, has the specific breeding of new brewing yeast strains become widespread. Strains have been produced with the ability to ferment a wider range of carbohydrates, with altered flocculation properties and which produce beers with modified flavours. Many have been tested on the pilot scale and one, an amylolytic brewing yeast, has received approval for commercial use.
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  • 3
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    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 28 (1995), S. 237-245 
    ISSN: 0739-4462
    Keywords: covalent modification ; amino acid uptake ; cotransport ; leucine ; Lepidoptera ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The arginine-specific reagent, phenylglyoxal, decreases the initial rate of lysine/K+ symport (cotransport) as well as maximum lysine accumulation at pH 9.2, by brush border membrane vesicles obtained from the larval midgut of the lepidopteran, Manduca sexta. The symport of a neutral amino acid, leucine, remained unaffected. Following exposure to phenylglyoxal, the apparent dissociation constant for lysine increased by a factor of 2.5 whereas the maximum uptake rate decreased by a factor of 0.4. More than one arginine residue appears to react with phenylglyoxal. Apparently phenylgyoxal reacts preferentially with arginine residues on a symporter that is specific for positively charged lysine. Phenylglyoxal shows promise as a specific covalent label for the identification of a cationic amino acid symporter. © 1995 Wiley-Liss, Inc.
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  • 4
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    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 28 (1995), S. 311-311 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 5
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    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 28 (1995), S. 257-272 
    ISSN: 0739-4462
    Keywords: Lepidoptera ; manducin ; storage protein ; tobacco hornworm ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The role of arylphorin as a storage protein was studied using 14C-arylphorin. 14C-arylphorin was produced optimally by incubating one-half fat body from Manduca sexta fifth instar larvae at 22°C for 24 h, in 1 ml of medium containing amino acids at 25% of their physiological concentration with [U-14C]-phenylalanine (phe) provided initially without nonlabeled phenylalanine. Nonlabeled phe was provided after 1 h at 16% of its physiological concentration. The specific activity of 14C-arylphorin produced in vitro was 30 times greater than that generated in vivo. Injection of 14C-arylphorin into pharate adults was used to study the distribution of 14C-phe derived from this protein into 14CO2 and tissues for comparison with injection of free 14C-phe during the middle (days 6 to 12 pharate adult) and late (days 12 to 17 pharate adult) stages of adult development. Appearance of 14CO2 from 14C-arylphorin as compared to 14C-phenylalanine showed a slower time course during both the middle and late stages of development, in keeping with the time needed for degradation of the protein. In accord with faster phe turnover near the end of adult development, total 14CO2 production was greater and the retention of 14C in hemolymph and fat body was less compared to the middle stage of development regardless of whether 14C-arylphorin or 14C-phe was injected. In the middle stage of development, the appearance of 14C in the cuticle and head parts was greater, whereas incorporation into abdomen and thorax was less than during the late stage of development. Since the pattern of 14C distribution from 14C-arylphorin and 14C-phe was similar, one major function of arylphorin must be as a storage protein replenishing the supply of free amino acids used for synthesis of adult tissues. These results also suggest a limited contribution of M. sexta arylphorin to formation of the cuticle subsequent to day-6 pharate adult. © 1995 Wiley-Liss, Inc.
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  • 6
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    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 28 (1995) 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 7
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    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 28 (1995), S. 397-406 
    ISSN: 0739-4462
    Keywords: glutathione ; glutathione S-transferase ; isozymes ; house fly ; resistance ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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.
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  • 8
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 25-34 
    ISSN: 0739-4462
    Keywords: Drosophila virilis ; diaphorase-1 ; quinone reductase ; dicoumarol ; enzyme induction ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Drosophila diaphorase-1 (DIA-1) is an enzyme similar to mammalian DT-diaphorase and is inhibitied in vitro by dicoumarol. However, a ten-fold increase in DIA-1 activity was observed when third instar Drosophila virilis larvae were fed on a diet containing 0.1 M dicoumarol for 48 h. This induction was shown to be dose dependent and immunoprecipitation experiments with DIA-1 antiserum demonstrated an increase in the DIA-1 protein level in dicoumarol-treated larvae. The induction of DIA1 by dicoumarol was found to be blocked by actinomycin D, which suggests a transcriptional mechanism of regulation. The opposite effect of dicoumarol on DIA-1 in vitro vs. in vivo suggests that a metabolic conversion takes place after the ingestion of this compound by D. virilis larvae. © 1995 Wiley-Liss, Inc.
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  • 9
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 247-255 
    ISSN: 0739-4462
    Keywords: lectin ; agglutination ; lipopolysaccharide ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A hemagglutinin was identified and partially characterized in the crop of the bug Triatoma infestans. It agglutinated mouse and rabbit erythrocytes, and was strongly inhibited by bacterial lipopolysaccharides. SDS-PAGE and immunoblotting with specific antibodies identified a dominant protein with molecular mass of 20 kDa under reducing conditions. The protein seems to be synthesized in the crop epithelial cells and deposited on the inner crop surface. Its possible biological role is discussed. © 1995 Wiley-Liss, Inc.
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  • 10
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    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 28 (1995), S. 273-289 
    ISSN: 0739-4462
    Keywords: neuronal death ; programmed cell death ; 20-hydroxyecdysone ; tobacco hornworm ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Half of the neurons in the abdominal nervous system of the moth Manduca sexta die after adult eclosion. Two physiological signals regulate post-eclosion neuronal death in adult moths. The first is endocrine: a decline in blood ecdysteroids is necessary for the death of neurons in the segmental ganglia. The second signal, which is highly specific for a pair of motoneurons found at the posterior midline in each of the three unfused abdominal ganglia, originates in the nervous system. It is transmitted from the fused pterothoracic ganglion to abdominal ganglion A3 via the intersegmental connectives. To characterize the signal of neural origin, we have developed an in vitro bioassay for neuron-killing factors (“neurocidins”). Aqueous extracts of pterothoracic ganglia were prepared and applied to cultured ventral nerve cords. These extracts exhibited concentration-dependent effectiveness in killing motoneurons. The active component of the extract was heat-stable and protease-sensitive. Size fractionation studies suggested that the active component has a molecular mass between 10 and 30 kD. This is the first report of an endogenous neuron-killing protein from an insect nervous system. © 1995 Wiley-Liss, Inc.
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  • 11
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 291-309 
    ISSN: 0739-4462
    Keywords: juvenile hormone ; methoprene ; pyriproxyfen ; fat body ; locust ; binding protein ; receptor ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Juvenile hormone (JH) binding components from the fat body of the African migratory locust were analyzed in a search for a potential nuclear JH receptor. Biosynthetically prepared 10R[3H]JH III gave a high proportion of specific binding to isolated nuclei and extracted proteins; data obtained with the JH analogs, [3H]methoprene and [3H]pyriproxyfen, on the other hand, were obscured by abundant non-specific binding. The vast majority of the high affinity JH III binding activity present in cytosolic and nuclear extracts was due to a high molecular weight JH binding protein (JHBP) which has previously been identified in locust hemolymph. This protein has several chromatographic forms which interfered in the search for a nuclear JH receptor. When specific antiserum was used to remove JHBP from nuclear extracts, a novel JH binding activity (NBP) was detected. NBP could be separated from JHBP by precipitation with ammonium sulfate. NBP displayed a high affinity for JH III (Kd = 0.25 nM) and JH I and JH II competed strongly for JH III binding, whereas methoprene and pyriproxyfen showed apparent competition when present in 1,000-fold excess. NBP was present in nuclear extracts at approximately 25,000 sites per cell; levels were similar in male and female locusts and were not greatly affected by the presence or absence of JH. The characteristics of NPB make it a strong candidate for a nuclear JH receptor. © 1995 Wiley-Liss, Inc.
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  • 12
    ISSN: 0739-4462
    Keywords: monooxygenases ; metabolism ; lauric acid ; ECOD ; testosterone ; insecticide ; dodecenoic acid ; inactivation ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In vitro bioassays were used to analyze the metabolism of the 11-dodecenoic acid (11-DDNA) by microsomes prepared from Drosophila melanogaster RalDDTR strain. 11-DDNA is metabolized to 11,12-epoxylauric acid (epoxyLA) in a NADPH-dependent way. The microsomal production of epoxyLA reaches a plateau very quickly, suggesting the occurrence of an enzyme inactivation process. After incubation of microsomes with (1-14C)11-DDNA, three proteins of Mr ≍ 50 kDa were labeled. 11-DDNA inhibits the microsomal metabolism of lauric acid and 7-ethoxycoumarin in a time and NADPH-dependent process. An inhibition of metabolites generated from DDT and testosterone was also obtained but at higher concentrations. These results are discussed according to the fact that RalDDTR is an insecticide resistant strain characterized as a high metabolizer of the insecticide DDT and also of lauric acid, testosterone, and ethoxycoumarin. © 1995 Wiley-Liss, Inc.
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  • 13
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 387-396 
    ISSN: 0739-4462
    Keywords: Delia antiqua ; superoxide dismutase ; purification ; monoclonal antibody ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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.
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  • 14
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 377-385 
    ISSN: 0739-4462
    Keywords: hydrocarbons ; diapause ; puparium ; water conservation ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Hydrocarbons on the puparia of flesh flies, Sarcophaga crassipalpis, were analyzed to determine whether the abundance of hydrocarbons on puparia from diapausing individuals (twice the amount extracted from puparia of nondiapausing individuals) was the consequence of an increase in deposition of select hydrocarbons or an overall increase in deposition. Hydrocarbons from the puparia of both diapausing and nondiapausing individuals are saturated and range in chain length from 25 to 33 carbons. GC-MS analyses indicate that the hydrocarbon fraction contains n-, terminally and internally branched monomethyl-, and 3,x-, 5,x- and internally branched dimethylalkanes. The diapausing and nondiapausing empty puparia contained 39.4 and 42.9% n-alkanes, 46.5 and 44.7% monomethylalkanes, and 9.5 and 8.5% dimethylalkanes, respectively. No major differences in the percent composition of the different hydrocarbons were noted between the two groups. This suggests that the amount of hydrocarbon, rather than the composition, contributes to the lower transpiration rates observed in diapausing pupae. © 1995 Wiley-Liss, Inc.
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  • 15
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 1-9 
    ISSN: 0739-4462
    Keywords: Solenopsis invicta ; ants ; fungi ; sitosterol ; ergosterol ; cholesterol ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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.
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  • 16
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 35-44 
    ISSN: 0739-4462
    Keywords: PBAN ; fatty acyl precursor ; Manduca sexta ; sex pheromone ; aldehydes ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Manduca sexta females that were decapitated produced no pheromone during the scotophase following decapitation, indicating that they were free of pheromone biosynthesis activating neuropeptide (PBAN). When deuterated hexadecanoic or (Z)-11-hexadecenoic acid was applied to the sex pheromone glands of decapitated or intact females of the same age, and allowed to incubate in vivo for 24 h, deuterium labeled Δ-11- and Δ-10, 12-unsaturated 16-carbon fatty acids were produced in both types of females. Injection of PBAN into intact or decapitated females 23 h after application of labeled acids had no effect on the production of unsaturated labeled fatty acids. However, deuterium labeled aldehydes were produced only in females that were injected with PBAN. Therefore, in this species, PBAN activates the process by which fatty acyl precursors in the pheromone gland are converted into the pheromonal aldehydes. © 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.
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  • 17
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 99-99 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 18
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 135-157 
    ISSN: 0739-4462
    Keywords: arsenic ; dioxin ; dichlone ; doxorubicin ; mitomycin C ; chloranil ; mercury ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recently progress has been made on O2 toxicity and pathology related to numerous environmental contaminants in insects. The pro-oxidants studied included: dioxin, paraquat, and an assorted array of quinones, 8-methoxypsorlen, arsenic, and mercury. The responses to these oxidants are diverse, but they arise from the reactive oxygen species. These pro-oxidants in insects cause lipid peroxidation, protein and enzyme oxidation, and GSH depletion. Potentially, they may also cause DNA oxidation, and form DNA adducts. Oxidative challenge is alleviated by antioxidant compounds, but more importantly by the induction of antioxidant enzymes, which are crucial for the termination of O2 radical cascade and lipid peroxidation chain reaction. Insects exhibit a wasting syndrome under sub-acute stress. In acute toxicity vital physiological processes impaired are hemolymph melanization and diuresis. Thus, insects resemble vertebrates in both the response to oxidative stress and its pathological consequences. These results raise the prospect that insects may serve as non-mammalian model species for monitoring the oxidative-stress component of environmental toxicity. © 1995 Wiley-Liss, Inc.
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  • 19
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 211-226 
    ISSN: 0739-4462
    Keywords: antioxidants ; phototoxins ; glutathione ; lipid peroxidation ; detoxification ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Plants of the Asteraceae and Hypericaceae possess secondary compounds that induce photooxidation in insect herbivores that consume them. One of the well-established modes of action of these substances is peroxidation of membrane lipids. Some herbivores counteract these defences by avoidance of light and tissues rich in phototoxins or the ability to detoxify these secondary substances. The cytochrome P-450 polysubstrate monooxygenase systems involved, the metabolic products, and a new putative toxin pump have been described. Dietary antioxidants (β-carotene, vitamin E, ascorbate) are additional defences against phototoxicity. They reduce mortality in herbivores exposed to phototoxins and some specialist herbivores have high constitutive levels. Adapted specialist insects also have higher constitutive levels of superoxide dismutase (SOD) and respond to phototoxins in their diet by the induction of catalase (CAT), glutathione reductase (GR), and increased levels of reduced glutathione (GSH). Artificial inhibition of the enzymes SOD and CAT had little effect on phototoxicity but inhibition of GSH synthesis in herbivores enhanced photooxidative effects of administered phototoxins on lipid peroxidation. While insects have many mechanisms to overcome plant photooxidants, the Asteraceae appear to have adopted a strategy of counterattack. We suggest and provide preliminary evidence that a second group of secondary substances, the sesquiterpene lactones, occurring in the Asteraceae can attack key antioxidant defences to synergise phototoxins. © 1995 Wiley-Liss, Inc.
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  • 20
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 227-242 
    ISSN: 0739-4462
    Keywords: wheat bug ; vitellin purification ; electrophoresis ; chromatography ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A single vitellin was identified in the eggs of the wheat bug, Eurygaster integriceps, and purified by a two-step procedure including ion exchange and gel permeation chromatography. Its nondenatured molecular mass is estimated to be 385 kDa. Under reducing conditions - SDS-PAGE - the wheat bug vitellin gives five polypeptides at 110 kDa, 80 kDa, 69 kDa, 53 kDa, and 38 kDa. Its amino acid composition is characterized by high content of aspartic acid/asparagine and glutamic acid/glutamine and low content of methionine and histidine. The lipid moiety (5.65% by weight) includes diacylglycerols, cholesterol, phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin. Carbohydrate content amounts to 4.54% by weight. A part of the wheat bug vitellin is dimerized during the course of deposition into the yolk granules. © 1995 Wiley-Liss, Inc.
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  • 21
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 259-268 
    ISSN: 0739-4462
    Keywords: aphids ; sexual generation ; gynoparae ; oviparae ; male development ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In order to complete a holocycle of the green peach aphid, Myzus persicae, on a synthetic diet, the effects of dietary amino acid contents on development of sexuals were investigated. For this purpose, the aphids were reared on holidic diets for two to three generations at 20°C under a scotophase of 15 h per diem. Under these conditions, the production of males by the aphids on a synthetic diet was very much poorer than those reared on radish seedling. On reducing the amino acid concentration of the diet, the proportion of males produced was comparable to that produced by aphids fed on radish seedlings. The obtained males were smaller and survived longer than those reared on radish seedling. Under the long night photoperiod and on the synthetic diet with reduced amino acids, gynoparae and then oviparae were also obtained. The oviparae were mated with the males grown on the synthetic diet, and laid eggs. The eggs, however, did not turn dark in color, and perished. © 1995 Wiley-Liss, Inc.
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  • 22
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 293-307 
    ISSN: 0739-4462
    Keywords: ferritin ; cDNA sequence ; Aedes aegypti ; mosquito ; cloning ; expression ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ferritin, an iron storage protein, was isolated from larvae and pupae of Aedes aegypti grown in an iron-rich medium. Mosquito ferritin is a high molecular weight protein composed of several different, relatively small, subunits. Subunits of molecular mass 24, 26, and 28 kDa are equally abundant, while that of 30 kDa is present only in small amounts. The N-terminal sequence of the 24 and 26 kDa subunits are identical for the first 30 amino acids, while that of the 28 kDa subunit differs. Studies using antiserum raised against a subunit mixture showed that the ferritin subunits were present in larvae, pupae, and adult females, and were increased in animals exposed to excess iron.The antiserum also was used to screen a cDNA library from unfed adult female mosquitoes. Nine clones were obtained that differed only in a 27 bp insertion in the 3′ end. Rapid amplification of cDNA ends (RACE) was used to obtain the complete protein coding sequence. A putative iron-responsive element (IRE) is present in the 5′-untranslated region. The deduced amino acid sequence shows a typical leader sequence, consistent with the fact that most insect ferritins are secreted, rather than cytoplasmic proteins. The sequence encodes a mature polypeptide of 20,566 molecular weight, smaller than the estimated size of any of the subunits. However, the sequence exactly matches the N-terminal sequences of the 24 and 26 kDa subunits as determined by Edman degradation. Of the known ferritin sequences, that of the mosquito is most similar to that of somatic cells of a snail. © 1995 Wiley-Liss, Inc.
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  • 23
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 357-379 
    ISSN: 0739-4462
    Keywords: yolk proteins ; vitelline membrane ; immunofluorescent staining ; immunogold labeling ; Indianmeal moth ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The production and uptake of the follicular epithelium yolk protein (FEYP) is terminated coincident with the initiation of vitelline membrane synthesis in follicles of the Indianmeal moth, Plodia interpunctella (Hübner). This was determined by visualizing the cytolocalization of the FEYP subunits YP2 and YP4 using antisera to immunolabel ultrathin sections or whole-mounted ovaries. Both subunits of FEYP were detectable in the Golgi apparatus and associated secretory granules of the follicular epithelial cells (FC) in vitellogenic follicles. Before the follicles entered the terminal growth phase, the oocytes began production of specialized organelles, late yolk spheres. Following the appearance of late yolk spheres in the oocyte, the FC initiated the production of vitelline membrane proteins and the rapid clearance of YP2 from their cytoplasm. No YP2 was detected in the Golgi apparatus or in the secretory granules of FC from follicles in terminal growth phase, although YP4 was detected in these organelles. The vitelline membrane of follicles in termal growth phase was a bilayered structure with an electron-dense layer of vitelline membrane proteins that originated in the FC and an electron-translucent layer containing yolk proteins. During this period, late yolk spheres were observed fused with the oolemma exposing and possibly releasing their contents to the electron-translucent layer of the vitelline membrane. From this evidence, we suggest that during termination of vitellogenesis, the oocyte and FC work in concert to end uptake of yolk proteins and begin the synthesis of egg membranes, and that the oocyte contributes to the production of vitelline membrane by the release of previously sequestered yolk proteins. © 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.
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  • 24
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 391-396 
    ISSN: 0739-4462
    Keywords: cicada ; adipokinetic hormone ; peptide ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Two peptides related to locust adipokinetic hormone and crustacean red pigment concentrating hormone were isolated by high performance liquid chromatography from the cicadas Cacama valavata and Diceroprocta semicincta. Both species have the same peptides. The structure of one of the peptides is pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp-Gly-Asn-amide. The mass spectrum, amino acid composition, and amino acid sequence of the other peptide suggest that it is almost identical to the first peptide. However, the exact nature of the difference between the two peptides could not be determined. © 1995 Wiley-Liss, Inc.
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  • 25
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 415-430 
    ISSN: 0739-4462
    Keywords: juvenile hormone binding protein ; hemolymph ; Melanoplus sanguinipes ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the hemolymph of Melanoplus sanguinipes, a high molecular weight juvenile hormone binding protein (JHBP) was identified by photoaffinity labelling and found to have a Mr of 480,000. The JHBP, purified using native gel electrophoresis followed by electroelution, has an equilibrium dissociation constant for JH III of 2.1 nM and preferentially binds JH III over JH I. Antibody raised against JHBP recognized only the 480,000 band. Under denaturing conditions the native JHBP gave a single band with a Mr 78,000. The antibody against native JHBP recognized only the 78,000 protein in SDS-treated hemolymph samples, indicating that JHBP is a hexamer in this species. The concentration of JHBP fluctuates in both the sexes during nymphal and adult development in parallel with total protein content of hemolymph. © 1995 Wiley-Liss, Inc.
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  • 26
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 25-40 
    ISSN: 0739-4462
    Keywords: Acyrthosiphon pisum ; Aphid ; 2,2-dimethylchromenes ; Metamorphosis ; Precocenes ; Wing determination ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The relationship between the structure of nineteen 2,2-dimethylchromene derivatives and their effects on aphid morphogenesis were investigated in a pink clone of the pea aphid, Acyrthosiphon pisum (Harris). Three bioassay systems were used: (1) wing induction - the induction of winged (alate) progeny by winged adults that normally produce only wingless (apterous) daughters, (2) wing inhibition - the inhibition of production of winged progeny by wingless adults that had been crowd-induced to promote the appearance of winged progeny, (3) the effect on metamorphosis - the production of precocious adults indicating a decrease in juvenile hormone titre or the induction of supernumerary moults indicating a juvenile hormone agonist effect. Compounds demonstrating wing-promoting effects had short (≤2 carbon) side chains at the C6 and/or C7 positions while methylation of C5 tended to decrease this activity. Of the seven compounds inducing wing formation, three also inhibited the production of winged progeny. However, the compounds affecting metamorphosis, in particular promoting precocious adult development, were similar to those that promoted wing inhibition rather than those with wing inducing effects; they had alkoxy groups at C7 with lengths of ≥2 carbons.There is a stronger correlation between compounds interfering with metamorphosis (and therefore evidenced to be affecting juvenile hormone levels, a classic property of some 2,2-dimethylchromene derivatives) and the promotion of wingless forms than the induction of winged forms. This finding is in contradiction to the idea that juvenile hormones are involved in promoting wingless forms. In addition, attempts to reduce the wing-inducing properties of Precocene II (the most potent compound effecting wing induction) by subsequent treatment with juvenile hormone Ill or the juvenile hormone analogue, pyriproxyfen, were inconclusive and attempts to inhibit w ng formation with these two compounds atter crowding were also unsuccessful. The precise mode of action of the 2,2-dimethylchromenes in relation to aphid wing induction remains unclear but it seems likely that the effect is not related to changes in juvenile hormone titres. © 1095 WiIey-Liss, Inc.
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  • 27
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 307-319 
    ISSN: 0739-4462
    Keywords: PBAN ; Heliothis peltigera ; moths ; insect neuropeptide ; sex pheromones ; Lepidoptera ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A structure-activity relationship study of Hez-PBAN was performed with respect to its pheromonotropic activity, using Heliothis peltigera as the test animal. The activity of N- and C-terminally derived sequences was examined in a time- and dose-dependent mode. Using a variety of Hez-PBAN-derived fragments at two doses (1 and 10 pmol) and at different times post-injection (5-120 min), we were able to demonstrate that peptides lacking 12 and 16 amino acids from their N-terminus are as potent as the full length PBAN, and that the C-terminally derived hexapeptide was capable of stimulating sex pheromone production to a similar extent as PBAN 1-33NH2, when its activity was analyzed at shorter post-injection times. Within the C-terminal sequence, the amide was found to play a crucial role. In addition, it was observed that the region between amino acids 9 and 13 is important for the biological activity of the full length PBAN. The fact that the pheromonotropic activity of the hexapeptide was similar to that of the full length PBAN, under specific conditions, suggests that this sequence constitutes the biologically active site of the neuropeptide. The discovery that PBAN-derived peptides reacted in a time- and dose-dependent mode, strengthens the assumption that proteolytic enzymes interfere with the pheromonotropic activity of the PBAN-derived fragments. The ability of a variety of peptides to stimulate sex pheromone biosynthesis suggests two possible mechanisms: (1) Existence of multiple pheromonotropic mechanisms which may be mediated by multiple PBAN receptors that are activated at different kinetics; (2) Existence of only one mechanism mediated by short C-terminally derived peptides. In the first case, the C-terminally derived sequences fulfill the conformational requirement of only one class of receptors, and other regions in the PBAN molecule (e.g., 9-13) fulfill the conformational requirements of a second (or other) class of receptors. In the second case, the C-terminally derived sequence is the only conformationally important sequence, and other sequences, which were found to be essential for the biological activity, serve other non-conformational purposes (e.g., protection against proteolytic degradation). © 1995 Wiley-Liss, Inc.
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  • 28
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 383-400 
    ISSN: 0739-4462
    Keywords: juvenile hormone ; pyriproxyfen ; accessory gland ; locust, esterase ; binding protein ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The participation of juvenile hormone (JH) in the regulation of growth and protein synthesis in the accessory reproductive gland of male Locusta migratoria has been investigated. After elimination of endogenous JH with ethoxyprecocene, the accessory gland failed to grow, but growth was restored by a single application of the JH analog, pyriproxyfen. Pyriproxyfen appeared to stimulate total protein synthesis by 3 h, with a significant effect by 12 h, in contrast to 24 h observed in fat body. The dose curve for stimulation of protein synthesis 12 h after applying pyriproxyfen gave an ED50 of 0.1 μg; the dose curve for gland growth at 72 h was biphasic, with steps at about 0.01 μg and 10 μg, suggesting two phases in JH action. SDS-PAGE analysis showed several components that were stimulated by pyriproxyfen, the effect being strongest in an 11 kDa band. A 5 kDa component was enhanced in the soluble and reduced in the particulate fraction after precocene treatment. The accessory gland contained JH esterase activity at levels about 100 times those in fat body or hemolymph, and was higher in precocene treated locusts. Binding activity for [3H]10R-JH III was high in cytosolic and nuclear fractions, and was identified immunologically as due to the previously described hemolymph JH binding protein. The results indicate that the mode of action of JH in the accessory gland may differ from that in the fat body. The presence of intracellular JH binding protein suggests a direct action of JH within the gland, that may be modulated by JH esterase. © 1995 Wiley-Liss, Inc.
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  • 29
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 369-381 
    ISSN: 0739-4462
    Keywords: pheromone biosynthesis activating neuropeptide ; enzyme linked immunosorbent assay ; sex pheromone ; Spodoptera littoralis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A highly sensitive enzyme linked immunosorbent assay (ELISA) for the determination of the pheromone biosynthesis activating neuropeptide (PBAN) has been developed. Six antisera have been obtained that recognize the carboxyl terminal side of this peptide. Two immunogens have been rationally designed and synthesized in order to direct antibody specificity, using as haptens PBAN or PBAN(20-33) with a Cys residue attached to their amino-terminal side. The Cys thiol group has been used to covalently bind the peptide to keyhole limpet hemocyanin (KLH) by using N-succinimidyl-4-(maleidimidomethyl) cyclohexane carboxylate (SMCC) as a convenient heterobifunctional cross-linker. Several usable competitive immunoassays have been obtained by synthesizing eight different coating antigens and screening the sera against all of them. The best assay was obtained with antibody 4 using Cys-Hez-PBAN(20-33) coupled to bovine serum albumin (BSA) through the Lys groups by using the homobifunctional cross-linker dimethylpimelidate dihydrochloride (DMP) as the coating antigen. The optimized assay allows to detect PBAN at concentrations as low as 1 fmol/well (l50 = 2.5 fmol/well). An extraction procedure for the hemolymph has been developed that allows to perform PBAN measurements in this tissue even after a tenfold dilution. In these conditions matrix effect is negligible. Preliminary results on the presence of PBAN like immunoreactivity (PBAN-IR) in the hemolymph of Spodoptera littoralis females are reported.© 1995 Wiley-Liss, Inc.
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  • 30
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 115-132 
    ISSN: 0739-4462
    Keywords: tobacco hornworm ; metamorphosis ; 20-hydroxyecdysone ; nuclei ; RNA polymerases ; ELISA ; monoclonal antibodies ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The activities of RNA polymerases I and II in the fat body of the tobacco hornworm larva increased during feeding and then declined during wandering in the fifth instar (Sridhara and Gilbert, Dev Biol 45:7-21 [1975]). RNA polymerase II activity was much higher than that of RNA polymerase I. To determine how the differences in the activities of the two enzymes and their developmental changes correlate with changes in RNA and protein, the contents of these macromolecules were determined during the same period. RNA and protein increased by about 8 to 10-fold during the feeding period. Activities of RNA polymerases I and II in nuclei, as well as those present in “free” (not transcribing) and “engaged” (transcribing) pools, were measured. Incorporation of labeled uridine into RNA by fat body cells in vitro was correlated to nuclear enzyme activities. The amounts of RNA polymerases I, II, and III were determined by the application of an ELISA with subunit-specific monoclonal antibodies. Results showed that even though the in vitro activity of RNA polymerase II and its amounts are higher than those of RNA polymerase I and III, rRNA and tRNA syntheses predominated over mRNA synthesis in vivo. The activities and amounts of RNA polymerase II in the “free” pool were higher than those for RNA polymerase I. The “free” pools of the two enzymes increased as the larva progressed through the fifth instar. There appears to be little correlation between RNA polymerase activities and RNA synthesis during wandering, as RNA synthesis by the tissue and nuclear RNA polymerase activities declined faster than the amounts of the enzymes. The amount of RNA remained fairly steady during the same period. 20-hydroxyecdysone did not affect either the RNA polymerase activities or RNA synthesis of the fat body cultured in vitro. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 45-69 
    ISSN: 0739-4462
    Keywords: Homoptera ; Aphididae ; amino acid metabolism ; symbiosis ; radioactive labeling ; essential amino acids ; glutamate ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Aphids are highly specialized insects that feed on the phloem-sap of plants, the amino acid composition of which is very unbalanced. Amino acid metabolism is thus crucial in aphids, and we describe a novel investigation method based on the use of 14C-labeled amino acids added in an artificial diet. A metabolism cage for aphids was constructed, allowing for the collection and analysis of the radioactivity incorporated into the aphid body, expired as CO2, and rejected in the honeydew and exuviae. This method was applied to the study of the metabolism of eight energetic amino acids (aspartate, glutamate, glutamine, glycine, serine, alanine, proline, and threonine) in the pea aphid, Acyrthosiphon pisum. All these amino acids except threonine were subject to substantial catabolism as measured by high 14CO2 production. The highest turnover was displayed by aspartate, with 60% of its carbons expired as CO2. For the first time in an aphid, we directly demonstrated the synthesis of three essential amino acids (threonine, isoleucine, and lysine) from carbons of common amino acids. The synthesis of these three compounds was only observed from amino acids that were previously converted into glutamate. This conversion was important for aspartate, and lower for alanine and proline. To explain the quantitative results of interconversion between amino acids, we propose a compartmentation model with the intervention of bacterial endosymbiotes for the synthesis of essential amino acids and with glutamate as the only amino acid supplied by the insect to the symbiotes. Moreover, proline exhibited partial conversion into arginine, and it is suggested that proline is probably indirectly involved in excretory nitrogen metabolism. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 87-97 
    ISSN: 0739-4462
    Keywords: cuticular lipid ; housefly ; hydrocarbon ; methyl-branched ; pheromone ; unsaturation ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We examined the biophysical properties of cuticular lipids isolated from the housefly, Musca domestica. Melting temperatures (Tm) of surface lipids isolated from female houseflies decreased from 39.3 °C to 35.3 °C as the females attained sexual maturity and produced sex pheromone, whereas those prepared from males did not change with age. Lipids melted over a 10-25 °C temperature range, and their physical properties were a complex function of the properties of the component lipids. The Tm of total cuticular lipids was slightly below that of cuticular hydrocarbons (HC), the predominant lipid fraction. Hydrocarbons were further fractionated into saturated, unsaturated, and methyl-branched components. The order of decreasing Tm was total alkanes 〉 total HCs 〉 methyl-branched alkanes 〉 alkenes. For 1-day-old flies, measured Tms of hydrocarbons were 1.3-5.5 °C lower than Tms calculated from a weighted average of Tms for saturated and unsaturated components. For 4-day-old flies, calculated Tms underestimated Tm by 11-14 °C. © 1995 Wiley-Liss, Inc.
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  • 33
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 71-85 
    ISSN: 0739-4462
    Keywords: oogenesis ; yolk proteins ; vitelline membrane ; immunofluorescent staining ; immunogold labeling ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Vitellin (Vt) was found not to be critical to the formation or structure of yolk spheres in oocytes of the moth, Plodia interpunctella (Hübner). Vitellogenic activities of the follicular tissues were determined by visualizing the immunocytolocalization of Vt subunits (YP1 and YP3) and of a follicular epithelium yolk protein (FEYP) subunit (YP2) in ultrathin sections or in whole-mounted tissues. Vitellogenin was detectable in the inter-follicular epithelial cell (FC) spaces of patent, vitellogenin follicles of normal females. When the follicles entered terminal growth phase, the inter-FC spaces closed equatorially around the follicle which excluded vitellogenin from that region. The closure of the spaces spread towards the poles in more mature follicles. Vt was immunolocalized to yolk spheres of vitellogenic and terminal growth phase oocytes. To examine the role of Vt in formation of yolk spheres, ovaries were transplanted into males. Vt was not detected in the inter-FC spaces, vitelline membrane, or yolk spheres of follicles from transplanted ovaries developing in males. However, the FEYP subunit YP2 was detected in the Golgi apparatus and secretory vesicles of columnar FC and in the yolk spheres of the oocytes from transplanted ovaries. During the late vitellogenic period, late yolk spheres appeared in the cortical region of the oocytes. In addition, YP2 was detected in the electron-translucent vitelline membrane of terminal growth phase follicles. We conclude that Vt is not required for the formation of yolk spheres or the electron-translucent layer of vitelline membrane. © 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.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 119-134 
    ISSN: 0739-4462
    Keywords: phototoxicity ; plant-herbivore interactions ; photosensitization ; detoxification ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Phototoxic chemicals produced by plants have been studied in a number of contexts, most notably as protective agents against mammalian and insect herbivores. Although there are commonalities in the responses of these two groups of herbivores to plant phototoxins, there are differences as well. Whereas a greater range of chemical classes has been demonstrated to display phototoxicity against insects, considerably more information is available on symptomology of phototoxicity and mechanisms of action in mammals. The commonalities include alterations in behavior following ingestion, notably photophobia, disruption of integumentary integrity following contact or ingestion, and metabolic detoxification following ingestion, in the case of furanocoumarins involving cytochrome P450 monooxygenases. Not yet known to exist in insects are phototoxin-mediated effects on sensory (particularly visual) systems and phototoxicity resulting from abnormal chlorophyll metabolism. In order to gain greater understanding of the ecological significance of phototoxin-mediated plant defense against both insects and mammals, there is a need for more studies centered on natural associations. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 175-186 
    ISSN: 0739-4462
    Keywords: peroxy radicals ; reactive oxygen species ; antioxidants ; polyphenols ; alkaloids ; carotenoids ; singlet oxygen ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Many reactive oxygen species such as ozone, singlet oxygen, hydroxyl radical, and organic oxyradicals have been implicated in damage to plant organs and biopolymers such as chloroplasts, cell membranes, proteins, and DNA. The principal defenses against these reactive molecules and free radicals in plants include detoxifying enzymes (catalase, superoxide dismutase, etc.) and also lower molecular weight secondary products with antioxidant activity. These latter compounds include a great variety of phenolic compounds, carotenoids, nitrogenous, and sulfur-containing materials. Some of the more important mechanisms of action of the secondary compounds will be discussed, with emphasis on the use of structural and kinetic data to identify the most effective antioxidants against peroxy radical-induced damage, which is perhaps the most important of the oxidative stresses present in the usual environment of plants. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 187-197 
    ISSN: 0739-4462
    Keywords: ascorbic acid ; dehydroascorbic acid reductase ; carotenoids ; catalase ; ferritin ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Insects possess a suite of antioxidant enzymes and small molecular weight antioxidants that may form a concatenated response to an onslaught of dietary and endogenously produced oxidants. Antioxidant enzymes such as superoxide dismutase, catalase, glutathione transferase, and glutathione reductase have been characterized in insects. Water-soluble and lipid-soluble antioxidants such as ascorbate, glutathione, tocopherols, and carotenoids have not been well studied in insects but may play very important antioxidant roles. Additionally, the peritrophic matrix and trehalose may possess important antioxidant functions in insects. The enzymatic recycling of ascorbate, first noted in green plants, may also exist in insects. A greater understanding of these antioxidant systems may provide greater understanding about the ecological relationships of insects with their hosts. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 199-209 
    ISSN: 0739-4462
    Keywords: arsenate ; arsenite ; arsenic toxicity ; oxidative stress ; antioxidant enzymes ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The potential usefulness of an insect model to evaluate oxidative stress induced by environmental pollutants was examined with trivalent arsenic (As3+, NaAsO2) and pentavalent arsenic (As5+, Na2HAsO4) in adult female house flies, Musca domestica, and fourth-instar cabbage loopers, Trichoplusia ni. M. domestica was highly susceptible to both forms of arsenic following 48 h exposure in the drinking water with LC50s of 0.008 and 0.011% w/v for As3+ and As5+, respectively. T. ni larvae were susceptible to dietary As3+ with an LC50 of 0.032% w/w but seem to tolerate As5+ well with an LC50 of 0.794% concentration after 48 h exposure. The minimally acute LC5 dose of both As3+ and As5+ varied considerably but averaged 0.005% for both insects. The potential of both valencies of arsenic for inducing oxidative stress in the insects exposed ad libitum to approximately LC5 levels was assessed. The parameters examined were the alterations of the antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), glutathione transferase (GST), the peroxidase activity of glutathione transferase (GSTPX), and glutathione reductase (GR), and increases in lipid peroxidation and protein oxidation. SOD (1.3-fold), GST (1.6-fold), and GR (1.5-fold) were induced by As3+ in M. domestica but CAT and GSTPX were not affected. As5+ had no effect on M. domestica. In T. ni, the antioxidant enzyme activities were not affected by As3+ except for SOD which was suppressed by 29.4% and GST which was induced by 1.4-fold. As5+ had no effect except the suppression of SOD by 41.2%. Lipid peroxidation and protein oxidation, which represent stronger indices of oxidative stress, were elevated in both insects by up to 2.9-fold. However, based on the antioxidant enzyme response to the arsenic anions, the mode of action of arsenic induced oxidative stress may differ between the two insects. Until this aspect is further clarified, evidence at this time favors the prospect of As3+ as a pro-oxidant, especially for M. domestica. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 243-257 
    ISSN: 0739-4462
    Keywords: pheromone biosynthesis ; aldehydes ; alcohols ; oxidase inhibitor ; piperonyl butoxide ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An oxidase that converts primary aliphatic alcohols into aldehydes was discovered in the cuticle of the sex pheromone gland and in the papillae anales on the tip of the abdomen of Manduca sexta females. Oxidase activity was not found in the epidermal cells of the pheromone gland where fatty acid precursors of the pheromonal aldehydes are found. This oxidase requires oxygen and water to function and appears to have a rather broad substrate specificity. The activity of the oxidase is reduced by the application of piperonyl butoxide, which also interferes with the PBAN induced production of the natural pheromone aldehydes. However, endogenous alcohols cannot be found in the pheromone gland. Thus, it is not yet clear whether or not the oxidase is involved in the terminal step of biosynthesis of the pheromone aldehydes in M. sexta females. © Wiley-Liss, Inc.This article is a U.S. Government work and, a such, is in the public domain in the United States of America.
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  • 39
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 329-342 
    ISSN: 0739-4462
    Keywords: malathion ; malathion carboxylesterase ; carbaryl ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Resistance to the organophosphate insecticide, malathion, in a strain of Culex tarsalis mosquitoes is due to increased activity of a malathion carboxylesterase (MCE). To determine whether resistance was due to a qualitative or quantitative change in the MCE, the enzyme was purified from both malathion-resistant and -susceptible mosquitoes. Enzyme kinetic measurements revealed that the two strains have one MCE in common, but resistant mosquitoes also have a unique MCE which hydrolyses malathion 18 times faster. Interestingly, this MCE does not hydrolyse α-naphthyl acetate, a substrate commonly used to detect increased levels of esterases in other organophosphate-resistant insects. Unlike the over-produced esterase of some related mosquito species, each MCE in C. tarsalis accounts for only a small fraction (0.015%) of the total extractable protein in either strain. Therefore, resistance in these insects is due to the presence of a qualitatively different enzyme, and not to a quantitative increase of a non-specific esterase. This study therefore demonstrates that the underlying biochemical mechanisms of insecticide resistance in one insect cannot necessarily be predicted from those of another, even closely related species. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 343-355 
    ISSN: 0739-4462
    Keywords: hypolipemia ; hypoglycemia ; glycogen phosphorylase ; L. migratoria ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Feeding starved adult migratory locusts, Locusta migratoria, caused decreases of hemolymph lipid concentrations and of the percentage of active fat body glycogen phosphorylase which suggested that a molecule(s) from the neurosecretory system or the midgut may have been released to regulate metabolism. Fat body phosphorylase was also inactivated after insects were transferred from 0 to 25 ° C. In adults with elevated hemolymph lipid levels after the injection of small doses of corpus cardiacum extract (CC), feeding did not induce a decrease in hemolymph lipid concentrations. It appears that the processes initiated by feeding could not override the effects of the continued presence of adipokinetic hormone(s) (AKHs) in the hemolymph or their long-term effects.Aqueous, methanolic, or ethanolic extracts of brains or storage lobes (SL) of fed locust CC did not lead to decreases of hemolymph lipid concentrations. Bovine insulin was equally inactive when tested at doses which were previously reported to reduce lipid levels. Fractions of ethanolic brain extracts from 3-day-starved males collected after high-performance size-exclusion chromatography, however, produced hypoglycemic effects in fed males. Two biologically active fractions were found, one with high (≥ 10 kDa) and one with low molecular weight (approximately 1 kDa). © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 431-431 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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  • 42
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 1-23 
    ISSN: 0739-4462
    Keywords: Lepidoptera ; oviposition ; attractant ; repellent ; stimulant ; deterrent ; host-marking pheromone ; oviposition-deterring pheromone ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Phytochemical mediators serving as attractants, repellents, stimulants, or deterrents in oviposition behavior of moths and butterflies are reviewed in regard to the chemical mechanism of host selection. Ovipositing females seem generally to utilize plant volatiles as cues for orientation to host plants, and the subsequent contact evaluation of plants by means of less- or non-volatile secondary metabolites is usually of great significance in host recognition. Most lepidopterans appear to be induced to oviposit in response to a single host-specific compound, while extreme synergism of multiple components features the stimulatory system of oviposition enacted by some butterflies. Recent investigations clearly demonstrate that acceptance or rejection of a particular plant by females is regulated not only by the presence or absence of oviposition stimulants but by negative stimuli evoked by co-occurring deterrents. The epideictic pheromones implicated in host assessment by females are also referred to in this review. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 77-91 
    ISSN: 0739-4462
    Keywords: Cancer antennarius ; cholesterol ; ecdysteroids ; high-density lipoproteins ; receptors ; Y-organs ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Y-organs are the ecdysial glands of crustaceans, responsible for synthesis and secretion of ecdysteroid hormones. For this purpose, the glands acquire cholesterol as obligate precursor entirely from circulating high-density lipoprotein (HDL). A preceding study provided evidence for the mechanism of acquisition: Y-organs take up cholesterol bound to HDL by an energy-requiring process, receptor-mediated absorptive endocytosis. The present study characterized the receptors involved utilizing isolated Y-organ membranes. HDL binding was saturable and specific; a dissociation constant (Kd) of 1.08 × 10-7 M and a binding maximum at equilibrium (Bmax) of 70 μg HDL protein/mg membrane protein, were obtained. Binding was decreased by protease and was dependent upon calcium. Y-organs are regulated negatively by a peptide hormone from the eystalks, molt-inhibiting hormone (MIH). Y-organ membranes from de-eyestalked crabs (MIH absent) exhibited the same Kd value as membranes from intact crabs, but a Bmax 17% higher. Thus, MIH activity apparently does not change the binding affinity of HDL, but decreases the number of binding sites. These results agree with our previous findings that MIH depresses ecdysteroid synthesis in part by inhibiting cholesterol uptake. Generally, Y-organ cells appear to contain receptors for HDL that are of high affinity and high binding capacity, similar to the characteristics reported for the binding of insect HDL (vitellogenin) to fat bodies and oocytes. © 1995 Wiley-Liss, Inc.
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  • 44
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 337-350 
    ISSN: 0739-4462
    Keywords: lipid carrying protein ; lipid transport ; insect lipoprotein ; midgut ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 32P-Labelled midguts (32P-midguts) of Rhodnius prolixus females were incubated in the presence of nonradioactive purified lipophorin and the release of radioactivity to the medium was analysed. The radioactivity found in the medium was associated with lipophorin phospholipids. When the 32P-midguts were incubated in the absence of lipophorin, no 32P-phospholipids were found in the medium. Comparative analysis by thin-layer chromatography of 32P-phospholipids derived from metabolically labelled 32P-midgut or lipophorin particles after incubation with 32P-midgut showed some differences, revealing a possible selectivity in the process of phospholipids transfer. The transfer of phospholipids to lipophorin was linear with time up to 45 min, was saturable with respect to the concentration of lipophorin, and was half-maximal at about 5 mg/ml. The binding of 32P-lipophorin to the midgut at O°C reached the equilibrium at about 1 h of incubation. The binding of 32P-lipophorin was inhibited by an excess of nonradioactive lipophorin, which suggests a specific receptor for lipophorin. The capacity of midguts and fat bodies to transfer phospholipids to lipophorin varied during the days following the meal. When lipophorin enzymatically depleted of phospholipids by treatment with phospholipase A2 was incubated with 32P-midguts, the same amount of phospholipids was transferred, indicating a net gain of phospholipids by the particle. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 133-147 
    ISSN: 0739-4462
    Keywords: juvenile hormone ; methoprene ; hormone receptor gene ; insecticide resistance ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Adult functions of juvenile hormone (JH) have been described for Drosophila melanogaster and other dipteran insects, but preadult function for this hormone remains largely unknown in this order of insects. We have identified a mutation of Drosophila, Methoprene-tolerant (Met), which appears to alter JH reception during late larval development. The molecular cloning of Met will be a step toward understanding this gene and possibly identifying a preadult role(s) for JH. Molecular cloning was initiated using the technique of transposon-tagging with a transposable P element. P-element insertional alleles of Met were generated, and genomic libraries were constructed from two of these alleles. From these libraries P-element-bearing clones were isolated that in situ hybridized to the cytogenetic region where Met had been previously localized by genetic methods. Two of the alleles were shown to have complete P-elements inserted in similar, but not identical, locations in the predicted cytogenetic region where Met is located. A late-larval cDNA library was screened to identify transcriptional units in this region, and clones were recovered with homology to a DNA fragment abutting the P-element insertion site. These clones may represent Met cDNA molecules. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 177-194 
    ISSN: 0739-4462
    Keywords: juvenile hormone esterase ; Autographa californica nuclear polyhedrosis virus ; Heliothis virescens ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Baculovirus insecticides are receiving renewed attention as insect pest control agents following the development of fast-acting recombinant baculoviruses. Here we report on the construction and biological activity of a recombinant baculovirus derived from the nuclear polyhedrosis virus of Autographa californica which expresses a modified form of juvenile hormone esterase (JHE). The serine at the catalytic site of the JHE has been mutated to a glycine residue so that the protein does not degrade JH. The recombinant baculovirus expressing this modified form of JHE, named AcJHE-SG, has enhanced activity against lepidopteran larvae. Lethal times of the recombinant are 20 to 30% lower than for the wild type virus, and a 66% reduction in feeding damage caused by infected larvae is observed. This result is comparable to the best recombinant baculovirus developed to date, AcAaIT, which expresses an insect-selective scorpion toxin. The potential of these recombinant viruses for commercialization as insecticides is discussed. Bioassays of AcJHE-SG in conjunction with anti-JH agents indicate that the virus is not killing by an anti-JH mechanism. Larvae apparently die from contraction-paralysis, or disruption of the normal sequence of events at the molt. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 211-223 
    ISSN: 0739-4462
    Keywords: apoLp-III ; amino acid residues ; lipid binding activity ; M. sexta ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Chemical modification procedures were employed to neutralize charged amino acid side chains of apolipophorin III (apoLp-III). Glutamate plus aspartate carboxylate side chains were amidated while, in other experiments, the ε-amino groups of lysine residue side chains were acetylated. Circular dichroism (CD) spectroscopy was performed to assess the effect of chemical modification on the secondary structure of apoLp-III. Compared to control, unmodified apoLp-III, both amidated and acetylated apoLp-IIIs possessed significantly diminished levels of α-helical structure. A similarly significant amount of α-helix structure could be induced in both modified apoLp-IIIs, however, by the addition of 50% trifluoroethanol, a helix inducing solvent, indicating that the proteins have retained their capacity to form helical secondary structures. The lipid binding interactions of chemically modified apoLp-IIIs were also examined in lipoprotein binding assays. Whereas control, unmodified apoLp-III displayed lipid binding activity, neither modified apoLp-III was capable of interaction with the substrate lipid surface. In phospholipid binding assays using the model compound, dimyristoylphosphatidylcholine, acetylated apoLp-III failed to interact while amidated apoLp-III showed limited interaction. When sodium dodecyl sulfate (SDS) micelles were employed as a model lipid surface, interaction of the modified apoLp-IIIs was observed. To characterize the relative stability of the interaction of control and modified apoLp-IIIs with SDS micelles, urea denaturation studies were performed. These experiments showed that, while control and amidated apoLp-IIIs were relatively resistant to urea induced denaturation, acetylated apoLp-III was susceptible. Taken as a whole, the results suggest that charged amino acid residues play an important role in stabilization of the lipid-free helix bundle conformation of apoLp-III and may promote stabilization of the lipid bound state through charge-charge interactions with lipoprotein surface components. © 1995 Wiley-Liss, Inc.
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  • 48
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    Keywords: juvenile hormone kinase ; tritiation ; HPLC ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Juvenile hormone epoxide hydrolase (JHEH) and juvenile hormone diol phosphotransferase (JHDPT) were characterized from the Malpighian tubules of day 1 fifth instar Manduca sexta. An improved RP-HPLC assay is described for the major metabolites of (10R, 11S) juvenile hormone I: diol, acid, aciddiol, and diol-phosphate. JHEH is strictly associated with membrane fractions, while JHDPT is cytosolic. JHEH may be solubilized in active form by the nonionic detergents Thesit or MEGA-8. Separation of Malpighian tubule cytosol proteins using preparative isoelectric focusing yields two zones which contain JHDPT activity, at pl 4.8-5.1 and 6.8-8.2. The partially purified JHDPT from either zone requires both ATP and Mg2+ for activity, so this enzyme may be formally called either ATP:juvenile hormone diol phosphotransferase or juvenile hormone diol kinase (EC 2.1.7.3.). Metabolites more polar than JH I aciddiol and JH I diol-phosphate are generated in vivo from either [3H]JH I or [3H]JH I diol. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995) 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 313-324 
    ISSN: 0739-4462
    Keywords: adipokinetic hormone ; cockroach ; hypertrehalosemic hormone ; trehalose synthesis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An in vitro bioassay suitable for routine use to investigate hypertrehalosemic hormone (HTH)-dependent trehalose biosynthesis was developed for the cockroach fat body. Blaberus discoidalis fat bodies were isolated and divided so that eight matched pieces from a single tissue could be compared for multiple control and experimental treatments. Optimum incubation conditions and the properties of HTH-dependent trehalose synthesis were determined. Dose-response studies determined an EC50 of 0.044 nM HTH for male fat body and 0.16 nM HTH for female tissue. HTH increased trehalose production by male fat body 3-fold compared to only a 67% maximum increase by the female tissue, and only the male tissue was used in subsequent studies. Fat body required only 5-min exposure to HTH for maximum trehalose production for 1 h. Trehalose synthesis was inhibited by ≥ 15 mM trehalose in the incubation medium. The fat body showed a developmental increase in trehalose synthesis in vitro that was reflected by hemolymph trehalose in vivo. Basal and HTH-related trehalose synthesis were low between days 0 and 10, increased 3-fold by day 20, and were high thereafter. These studies have established baseline data for future investigations to identify the signal transduction mechanisms involved in HTH regulation of fat body metabolism. © 1995 Wiley-Liss, Inc.
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  • 51
    ISSN: 0739-4462
    Keywords: serine proteinases ; proteinase inhibitors ; trypsin ; Coleoptera ; Scarabaeidae ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The proteinases in the midguts of three scarab white grub species, Lepidiota noxia, L. negatoria, and Antitrogus consanguineus, were investigated to classify the proteinases present and to determine the most effective proteinase inhibitor for potential use as an insect control agent. pH activity profiles indicated the presence of serine proteinases and the absence of cysteine proteinases. This was confirmed by the lack of inhibition by specific cysteine proteinase inhibitors. Trypsin, chymotrypsin, elastase, and leucine aminopeptidase activities were detected by using specific synthetic substrates. A screen of 32 proteinase inhibitors produced 9 inhibitors of trypsin, chymotrypsin, and elastase which reduced proteolytic activity by greater than 75%. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995) 
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 11-23 
    ISSN: 0739-4462
    Keywords: cricket ; HPLC ; neuropeptide ; diuretic ; antidiuretic ; Malpighian tubule ; fluid secretion ; Mas-DP1 ; achetakinin ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the Malpighian tubules of Acheta, the distal and middle segments are functionally and morphologically quite distinct (Spring and Kim, Mol Comp Physiol 12:130-145, 1993). Furthermore, they respond quite differently to corpora cardiaca (CC) homogenates, dibutyryl cAMP, and A23187 (Kim and Spring, J Insect Physiol 38:373-381, 1992). In this study we compared secretion by these two regions in response to Acheta and Romalea CC extracts, synthetic Manduca sexta diuretic peptide (Mas-DP1), and the family of synthetic myotropic peptides, the achetakinins, isolated from Acheta. Both Acheta and Romalea CC extracts had opposite effects on the two regions: mid-tubule secretion increased 3-fold whereas secretion by the distal segment declined 75-80%. Mas-DP1 increased secretion by the mid-tubule more than 3-fold and had no effect on the distal segment. All of the achetakinins decreased secretion by the distal tubule, with achetakinin 1 being least effective (55% inhibition) and achetakinin 5 being most effective (75% inhibition). Achetakinins 1 and 2 increased midtubule secretion by 3.7- and 3.3-fold, respectively, whereas the others had no effect on this region. Regarding HPLC fractions of CC extracts, in general the more hydrophilic fractions inhibited secretion by both distal and mid-tubules. The more hydrophobic fractions were nearly uniformly stimulatory when applied to the mid-tubule, and either inhibited secretion or had no effect on the distal region. The possible interpretations of these data and the implications towards future research are discussed. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995) 
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 101-118 
    ISSN: 0739-4462
    Keywords: oxidative stress ; reactive oxygen species ; lipid peroxidation ; protein oxidation ; DNA oxidation ; antioxidants ; oxidant repair enzymes ; redox-active flavonoids ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The survival of all aerobic life forms requires the ground-state of molecular oxygen, O2. However, the activation of O2 to reactive oxygen species (ROS) is responsible for universal toxicity. ROS are responsible in deleterious intracellular reactions associated with oxidative stress including membrane lipid peroxidation, and the oxidation of proteins and DNA. Redox-active allelochemicals such as quinones and phenolic compounds are involved in activating O2 to its deleterious forms including superoxide anion free radical, \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm O}_{\rm 2} ^{ \cdot - } $\end{document}, hydrogen peroxide, H2O2, and hydroxyl radical, \documentclass{article}\pagestyle{empty}\begin{document}$ \cdot {\rm OH} $\end{document}. Molecular oxygen is also activated in biologically relevant photosensitizing reactions to the singlet form, 1O2. The insect lifestyle exposes them to a broad diversity of pro-oxidant allelochemicals and, like mammalian species, they have developed an elaborate antioxidant system comprised of chemical antioxidants and a bank of antioxidant enzymes. We have found that an insect's antioxidant adaptation to a particular food correlates well with its risk of exposure to potential pro-oxidants. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 159-173 
    ISSN: 0739-4462
    Keywords: superoxide dismutase ; catalase ; enzyme regulation ; oxidant ; anti-oxidant ; reactive oxygen species ; sigma factors ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sophisticated biochemical networks allow organisms such as bacteria and insects to switch from very rapid growth and development in ideal environments to dormancy during severely unfavorable conditions. These switches may be accompanied by abrupt changes in oxidation/reduction involving reactive oxygen species (ROS). ROS have the potential of damaging nucleic acids, proteins, and membranes. In Escherichia coli, certain genetically regulated circuits (regulons) turn on synthesis of anti-oxidant enzymes to protect against distinct ROS excesses (superoxide, hydrogen peroxide, organic or lipid peroxides, etc.). As examples, the soxRS regulon controls synthesis of Mn-superoxide dismutase, oxyR controls catalase HPI, rpoS positively regulates HPII, and fur regulates several oxidative reactions that involve iron uptake. Our studies have focused on the regulatory role of rpoS, known to be a sigma factor (σ38) that combines with RNA polymerase and is a regulator of those gene products needed to protect cells during dormancy. Since insect cells, during both active growth and dormancy, endure severe environments, analogous protective gene products may be induced. Examples are presented of insect anti-oxidant metabolism, including those involved in the aging process. In addition, we searched several DNA and protein sequence data banks to compare resemblances between anti-oxidant gene products of bacteria and insects. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 321-336 
    ISSN: 0739-4462
    Keywords: triacylglycerols ; pheromone aldehydes ; sex pheromone gland ; Manduca sexta ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Analysis by TLC and HPLC revealed that the triacylglycerols comprise the most abundant lipid class in the sex pheromone glands of Manduca sexta females. Also, conjugated olefinic acyl analogs of the major pheromone aldehydes occur principally in the triacylglycerols. The amount of triacylglycerols with conjugated diene acyl moieties significantly decreased when the period of pheromone production was extended by 7 h beyond the normal period of pheromone production by 3 injections of pheromone biosynthesis activating neuropeptide (PBAN) at 3 h intervals. This decrease indicates that the triacylglycerols stored in the gland may serve as major sources of pheromone precursors in the biosynthesis of the sex pheromone aldehydes. Furthermore, analysis of pheromone aldehydes and triacylglycerols in the gland from moths treated with PBAN showed that the proportions of the triacylglycerols with conjugated diene moieties were closely correlated with the proportions of aldehydes found in the same gland. This correlation suggests that the proportions of fatty acids bound to certain triacylglycerols regulates the proportions of aldehydes in biosynthesis of the pheromone blend in M. sexta. © 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.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 339-349 
    ISSN: 0739-4462
    Keywords: Epiblema scudderiana ; Eurosta solidaginis ; cold-hardy insects ; low temperature acclimation ; cryoprotectant biosynthesis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Changes in the activity of over 20 enzymes of intermediary metabolism in 15°C or -4°C acclimated goldenrod gall moth (Epiblema scudderiana) and gall fly (Eurosta solidaginis) larvae were measured. Increased activities of glyco-genolytic and hexose monophosphate shunt enzymes in cold-acclimated Epiblema scudderiana suggest a role for coarse control in the conversion of glycogen reserves into glycerol cryoprotectant synthesis. In Eurosta solidaginis, high glycogen phosphorylase activity with decreased activities of glycolytic enzymes may account in part for the temperature-dependent switch from glycerol to sorbitol synthesis in these larvae upon cold acclimation. Isoelectric focusing analyses of five enzymes in overwintering Epiblema scudderiana revealed transient mid-winter changes in the isoelectric points of phosphofructokinase and pyruvate kinase, suggesting seasonal changes in the phosphorylation state of these enzymes. A distinct developmental pattern of aldolase isozymes suggests a role for a new isozyme during overwintering or upon spring emergence. Regulation of metabolism by changes in enzyme activities is indicated for both larvae. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 365-375 
    ISSN: 0739-4462
    Keywords: ecdysone 20-monooxygenase ; allelochemicals ; enzyme induction ; microsomal monooxygenases ; fall armyworm ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The influence of dietary allelochemical on ecdysone 20-monooxygenase activity was studied in the fall armyworm, Spodoptera frugiperda (J.E. Smith). Feeding the indoles (indole-3-carbinol, indole-3-acetonitrile), flavonoids (flavone, β-naphthoflavone), monoterpenes (menthol, menthone, peppermint oil), and a coumarin (xanthotoxin) to the larvae stimulated midgut microsomal ecdysone 20-monooxygenase activity from 28 to 200% as compared with the controls. β-Naphthoflavone was the most potent inducer among those tested. Phenobarbital, a well-known cytochrome P450 inducer, also caused a 2-fold increase in the microsomal ecdysone 20-monooxygenase activity. Ecdysone 20-monooxygenase activity was 2.7-fold higher in the microsomal fraction than in the mitochondrial fraction isolated from larval midguts. Microsomal ecdysone 20-monooxygenase activity was highest in the fat body, followed by the midgut and Malpighian tubules. Tissue localization and enzyme inducibility were different between ecdysone 20-monooxygenase and xenobiotic-metabolizing cytochrome P450 monooxygenases, including aldrin epoxidase, biphenyl hydroxylase, methoxyresorufin O-demethylase, 7-ethoxycoumarin O-deethylase, p-chloro-N-methylaniline N-demethylase, and phorate sulfoxidase in fall armyworm larvae. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 17-32 
    ISSN: 0739-4462
    Keywords: Calliphora ; tyrosine hydroxylation ; dopa oxidation ; dehydrodopa ; quinone reactivity ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Incubation of N-acetyltyrosine methyl ester with cuticular enzymes, isolated from the wandering stages of Calliphora sp larvae, resulted in the generation of N-acetyldopa methyl ester when the reaction was carried out in the presence of ascorbate which prevented further oxidation of the o-diphenolic product. Enzymatic oxidation of N-acetyldopa methyl ester ultimately generated dehydro N-acetyldopa methyl ester. The identity of enzymatically produced N-acetyldopa methyl ester and dehydro N-acetyldopa methyl ester has been confirmed by comparison of the ultraviolet and infrared spectral and chromatographic properties with those of authentic samples as well as by nuclear magnetic resonance studies. Since N-acetyldopaquinone methyl ester was also converted to dehydro N-acetyldopa methyl ester and tyrosinase was responsible for the oxidation of N-acetyldopa methyl ester, a scheme for the cuticular phenoloxidase catalyzed conversion of N-acetyltyrosine methyl ester to dehydro N-acetyldopa methyl ester involving the intermediary formation of the quinone and the quinone methide is proposed to account for the observed results. The conversion of N-acetyldopa methyl ester to dehydro derivative remarkably resembles the conversion of the sclerotizing precursor, N-acetyldopamine, to dehydro-N-acetyl-dopamine observed in the insect cuticle. Based on these comparative studies, it is proposed that peptidyl dopa derivatives could also serve as the sclerotizing precursors for the sclerotization of the insect cuticle. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 63-78 
    ISSN: 0739-4462
    Keywords: fat body ; glycogen ; phosphorylase ; hemolymph ; glucose ; chitosan ; lipids ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 113-129 
    ISSN: 0739-4462
    Keywords: Hyphantria cunea ; vitellin ; vitellogenin ; yolk proteins ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Yolk proteins (YP1, YP2, and YP3) of the fall webworm, Hyphantria cunea, are of relatively low molecular weight. Yolk protein-2 (YP2) was purified from gel slices and by KBr density gradient ultracentrifugation followed by ion exchange chromatography. YP2 is composed of one subunit with a molecular weight of 35.5 kDa. YP2 contains neutral lipids (diacylglycerol and triacylglycerol) and phospholipids (phosphatidylcholine and phosphatidylethanolamine). The neutral lipids are largely composed of lauric acid and palmitoleic acid. YP2 contains relatively large amounts of glutamic acid and aspartic acid but small amounts of tyrosine, phenylalanine, and methionine. YP2 is a vitellin (Vn) synthesized by the fat body. Vitellogenin-2 (Vg2), the precursor of YP2, is present in very small amounts in the hemolymph. Lipophorin and storage protein also are found in the ovary of H. cunea, and these proteins do not immunologically cross-react with YP2. YP2 is detected in first instar larvae but completely disappears during the second instar, indicating that YP2 is intensively utilized during postembryonic development. Anti-YP2 antibodies cross-react with ovarial extracts of Bombyx mori but not with those of insects from other orders such as Cletus schmidti (Hemiptera), Lucilia illustris (Diptera), Anechura japonica (Dermaptera), Periplaneta americana (Dictyoptera), and Ducetia japonica (Orthoptera). © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 159-171 
    ISSN: 0739-4462
    Keywords: myotropin ; myoactive peptide ; Locusta migratoria oviduct bioassay ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A five-residue myotropic peptide, Manduca sexta midgut myotropin I (Mas-MG-MT I), was isolated from an extract of 800 midguts of fifth instar larvae of the tobacco hornworm, Manduca sexta. It was purified by reverse phase and normal phase HPLC. Myotropic activity was screened by a heterologous Locusta migratoria oviduct bioassay. Sequence analysis, amino acid composition analysis, and comparison of candidate synthetic peptides in the amide and acid form revealed the following primary structure: Ala-Glu-Pro-Tyr-Thr-NH2. This is the first fully identified peptide isolated directly from the midgut of an insect species. Few significant sequence homologies with known vertebrate and invertebrate peptides have been found. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 209-223 
    ISSN: 0739-4462
    Keywords: RH-5849 ; RH-5992 ; Dibenzoyl hydrazine ; tebufenozide ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The first non-steroidal ecdysteroid agonists are dibenzoyl hydrazines and are typified by the compounds designated RH-5849 and RH-5992. The discovery that these compounds mimic 20E in a variety of insect orders, and especially the Lepidoptera, generated great interest from the research and agricultural communities. Such compounds provide important new research tools for physiological, biochemical, and molecular studies. In addition, the potential for application to agricultural pests looks very promising, especially for RH-5992 (tebufenozide). This review evaluates the evidence on the specificity of the ecdysteroid-like actions of these materials and considers their application for research and pest management. © 1995 Wiley-Liss, Inc.This article is a US Government work and, as such, is in the public domain in the United States of Amerira.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 269-280 
    ISSN: 0739-4462
    Keywords: Manduca ; tobacco hornworm ; scolexin ; immunity ; insect defense ; hemolymph proteins ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The immune protein, scolexin, a bacteria-induced, larva-specific protein from Manduca sexta, was shown to exist in the hemolymph in two isoelectric forms designated herein as scolexin-1 and scolexin-2 (native Mr ∼ 72 kd). These two charge isomers appeared to share the same amino acid composition. Scolexin is composed of two subunits (peptide Mr ∼ 36 kd) that possess the same N-terminus. Scolexin-2 was subjected to glycosyl composition analysis, revealing the presence of galactose, glucose, mannose, xylose, and sialic acid residues. Hybridization of epidermal RNA with oligonucleotides deduced from the scolexin N-terminal sequence showed a continuous decline in mRNA following day 0 of the 5th larval instar. By employing in vitro protein labelling, it was found that organ cultures of the epidermis from immune larvae showed a greater ability over that of naive epidermal cultures to synthesize scolexin; these data reflected the inducible response seen in the hemolymph, and confirm other data indicating that the epidermis is an important site of scolexin biosynthesis. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 1-15 
    ISSN: 0739-4462
    Keywords: precocious metamorphosis ; precocious molt ; spermatogenesis ; JH ; ecdysteroid ; developmental program ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: KK-42 (1-benzyl-5-[(E)-2,6-dimethyl-1,5-heptadienyl]imidazole), administered by feeding, delayed the growth and development of nondiapause-bound and diapause-bound Ostrinia nubilalis larvae and increased the length of the instar. At doses of 80-240 ppm, 62-100% of nondiapause-bound fourth instars precociously pupated or remained as fourth instars, while 52-100% of diapause-bound fourth instars did not molt to the fifth instar. Injection of these nondiapause- and diapause-bound KK-42-fed fourth instars with ecdysone elicited a molt and resulted in the production of larval-pupal intermediates. When mature fourth instar controls were similarly injected, they molted into normal fifth instars. These results support the view that KK-42 delays/inhibits ecdysteroid production. Both eupyrene and apyrene spermiogenesis were prematurely initiated in nondiapause-bound fourth instars that were fed on medium containing 160 ppm KK-42. Fenoxycarb, a potent juvenile hormone mimic, rescued nondiapause-bound fourth instars from precocious pupation. All fenoxycarbtreated larvae either molted to the fifth instar or remained as fourth instars and eventually died. These results support the view that treatment with KK-42 inhibits JH production. When KK-42 treatment was begun in the third instar, a considerable number of nondiapause-bound and some diapause-bound third instars precociously molted to the fifth instar. There was a correlation between weight and the incidence of precocious molting in that third instars destined to skip the fourth instar attained a weight, as pharate fifth instars, of two to three times more than pharate fourth instar controls. Similarly, fourth instars that were destined to undergo precocious pupation attained a weight, as pharate pupae, that was approximately two times more than pharate fifth instar controls. More potent analogues of KK-42 may prove useful in controlling populations of 0. nubilalis by interfering with their growth, development, and metamorphosis. © 1995 Witey-Liss, Inc.This article is a US Government work and, as such, is, in the public domain in the United States of America.
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 131-141 
    ISSN: 0739-4462
    Keywords: V-ATPase ; ovarian follicles ; Manduca sexta ; vitellogenesis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A monoclonal antibody, directed against an H+ translocating V-ATPase of the midgut of Manduca sexta, has been used for immunolocalization studies in ovarian follicles and testes of Manduca sexta. In testes, no distinct staining above background levels was observed. In vitellogenic follicles, V-ATPase immunoreactivity first appears in the cytoplasm of the trophocytes and then in the oocyte, but by far the strongest reaction is present in the region of the oolemma during endocytosis. All types of follicle cells surrounding both the oocyte and the trophocyte compartments show a distinct positive reaction. In the cylindrical follicle cells surrounding the oocyte, the immunoreactivity is clearly restricted to the basal part. Our results suggest an important role for V-ATPase in vitellogenin uptake in Manduca, similar to that suggested on electro-physiological grounds in Hyalophora cecropia. © 1995 Wiley-Liss, Inc.
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 189-205 
    ISSN: 0739-4462
    Keywords: digestion ; trypsin ; chymotrypsin ; multiple feeding ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Two types of trypsin activity were detected in Anopheles albimanus, a constitutive and an inductive component, which have identical immunopatterns. The constitutive trypsin in synthesized shortly after eclosion and is retained in the midgut epithelial cells. The inductive trypsin is synthesized and released continuously after a blood meal has been ingested; maximal activities vary between 12 h and 18 h after a blood meal. Once digestion is completed, trypsin is excreted, but the constitutive trypsin level is restored within 24 h, before the next blood meal is taken. In A. gambiae, A. Stephensi, and A. quadrimaculatus, the constitutive trypsin component is also present, but at much lower levels. In A. albimanus fed multiple blood meals at 24 h intervals, trypsin oscillates at nearly maximal levels as long as blood is present in the midgut and depending on the ovarian status. Expression of the two trypsin components in A. albimanus was found to be independent of the neurosecretory system, but synthesis of the constitutive trypsin appears to require the presence of the corpora allata. In all species tested, chymotrypsin is secreted after a blood meal in a similar temporal pattern as trypsin, but it is never present before the blood meal. Reinvestigating several aedine species for the presence of chymotrypsin by using different substrates revealed measurable quantities in blood-fed females compared to earlier reports. Equally, aminopeptidase activity is present in all species tested and characterized by a constitutive component. Its activities follow different temporal patterns than the endopeptidases. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 225-235 
    ISSN: 0739-4462
    Keywords: cryoprotectant production ; insect cold-hardiness ; glycerol catabolism ; Epiblema scudderiana ; gluconeogenesis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Fructose 1,6-bisphosphatase (FBPase) from the larvae of the gall moth, Epiblema scudderiana, was purified to homogeneity with a final specific activity of 1.6 U/mg protein. The enzyme had a native molecular weight of 74.0 ± 6.5 kD and a subunit molecular weight of 37.6 ± 3.0 kD; the dimeric structure of the enzyme in this species is unusual. The pH optimum was 7.00 in imidazole buffer at 22°C and rose to 7.31 at 5°C. An Arrhenius plot of enzyme activity vs. temperature was linear with an activation energy of 91 ± 4.1 kJ/mol-1. Km values for FBPase decreased from 4.7 ± 0.34 μM at 22°C to 1.3 ± 0.05 μM at 5°C. No allosteric activators were identified, but the enzyme was inhibited by fructose 2,6-bisphosphate (F2,6P2), AMP, ADP, dihydroxyacetonephosphate, glycerol, and KCI. Inhibition by AMP and F2,6P2 increased at low temperature, and effects of these compounds may be key to preventing futile cycling of carbon at the FBPase/phosphofructokinase loci during the biosynthesis of glycerol cryoprotectant. Oppositely, glycerol clearance in the spring and reconversion into glycogen is promoted by interactions of temperature, inhibitors, and glycerol that promote FBPase activity: I50 values for AMP and F2,6P2 increase at 22°C (compared with 5°C), high glycerol levels override F2,6P2 inhibition of the enzyme, and deinhibitors (ATP, citrate) partially reverse AMP inhibition of the enzyme. © 1995 Wiley-Liss, Inc.
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 381-390 
    ISSN: 0739-4462
    Keywords: cell line ; development ; Manduca sexta ; apolipoprotein III ; polypeptide ; vesicles ; Trichoplusia ni ; wing discs ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An N-terminal amino acid sequence of a 16.9 kDa hemolymph polypeptide, “Vesicle Promoting Factor” (VPF) from Trichoplusia ni, revealed a high sequence homology (70%) with Manduca sexta apolipophorin-III. A polyclonal antibody developed against VPF, however, was not immunoreactive with either purified M. sexta or T. ni apolipophorin-III. Immunoblots of tissue homogenates of T. ni indicated that VPF was present in imaginal wing discs, central nervous system (CNS), silk glands, midgut and hemocytes from fifth instar larvae, and also in the IAL-TND1 cell line which can grow as either fluid-filled multicellular vesicles or multicellular aggregates. VPF was also detected immunologically in the hemolymph of adults of T. ni, and in hemolymph of adults and larvae of Galleria mellonella and Heliothis virescens. Testes, midgut, hemocytes, and wing discs, but not Malpighian tubules, of T. ni released VPF into tissue culture medium during a 3 h incubation period. © 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.
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    Archives of Insect Biochemistry and Physiology 30 (1995) 
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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  • 77
    ISSN: 0739-4462
    Keywords: ecdysteroid ; nuclear receptor ; Chironomus tentans ; polytene chromosomes ; antibodies ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Antisera were raised against different domains of a putative ecdysteroid receptor (cEcRH) of Chironomus tentans. All the antisera reacted with a 68,000 dalton protein exhibiting DNA binding properties. Additionally, we were able to demonstrate that the antisera immunoprecipitate protein which binds a radioactively labeled ecdysteroid (Ec), i.e., [3H]ponasterone A, with high specificity. These properties indicate that the antisera recognize specifically an endogenous ecdysteroid receptor protein (cEcR) in C. tentans cells and thus are suitable for the following quantitative and qualitative immunological and immunohistochemical investigations. The cellular level of cEcR varies during development, and it is particularly low in oligopausing larvae. In polytene chromosomes of prepupal salivary glands, cEcR is located at approximately 50 transcriptionally active loci. These loci include both early ecdysteroid (Ec)-inducible puff sites, such as the locus containing the gene coding for the homolog of the E75 protein in Drosophila melanogaster, as well as late Ec-inducible puff-sites. The latter group comprises a locus of a gene specifying the homolog of the D. melanogaster ultraspiracle protein. However, loci of genes coding for salivary gland secretory proteins (e.g., Balbiani ring forming chromosome regions) do not specifically react with the antisera. Thus, the developmental regulation of these genes is not directly controlled by Ec. Polytene chromosomes of oligopausing larvae show hardly any loci that contain cEcR. The few detected correspond, with few exceptions, to the most potent cEcR binding sites found in prepupae. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 165-176 
    ISSN: 0739-4462
    Keywords: Lepidoptera ; radioimmunoassay ; enzyme immunoassay ; equilibrium dialysis ; monoclonal antibody ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Numerous studies have demonstrated regulation of specific lepidopteran proteins by pharmacological doses of insect juvenile hormone (JH). In this study, topical application of a 1 pg dose of JH I to fourth stadium larvae of the black (bl) mutant strain of the tobacco hornworm, Manduca sexta, induced a 50% increase in the titer of hemolymph juvenile hormone binding protein (hJHBP). Radioimmunoassay confirmed that JH titers were lower in bl larvae than in wild-type larvae at the time of JH treatment. Enzyme immunoassay analysis of hJHBP titers demonstrated that regulation by JH I was dose-dependent at doses up to 10 pg and that the response was saturated above 100 pg. Western blotting and equilibrium dialysis confirmed these results and demonstrated that hJHBP from bl larvae had the same molecular mass and displayed the same affinity for JH I as hJHBP isolated from wild-type larvae. Time course studies showed that regulation was complex: 1 2 h after JH I treatment, hJHBP titers were twofold lower in treated than in control bl larvae, while 44 h after treatment they were twofold higher. JH I regulation of hJHBP titers in bl larvae was independent of changes in total hemolymph protein. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 49-62 
    ISSN: 0739-4462
    Keywords: octopamine ; 5HT ; oviducts ; pharmacology ; Periplaneta americana ; visceral muscle ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effects of the amines 5HT and octopamine on the myogenic activity of Periplaneta americana (L.) oviducts and the pharmacological profile of octopamine and 5HT receptors on the lateral oviducts have been determined. Application of 5HT to the oviducts resulted in a dose-dependent increase in basal tonus and amplitude of contractions. Antagonist studies revealed that the 5HT receptor on the cockroach oviduct most resembles the mammalian 5HT2 receptor. Application of octopamine resulted in a decrease in basal tonus and had a biphasic effect on the amplitude of contractions, being stimulatory at low doses and inhibitory at higher ones. The inhibitory effects of octopamine appear to be mediated via cAMP and are blocked by antagonists which indicate that the octopamine receptor is of the octopamine-2 type. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 33-48 
    ISSN: 0739-4462
    Keywords: tobacco budworm ; parasitoid ; Braconidae ; Noctuidae ; Hymenoptera ; Lepidoptera ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Polydnaviruses from certain parasitoid Hymenoptera have been reported to interfere with both host immunity and host development. Heliothis virescens larvae injected with either calyx fluid or sucrose gradient-purified polydnavirus from Microplitis croceipes (McPDV) gained less weight than saline-injected larvae. The active feeding portion of the fifth stadium larva (time to reach the burrowing-digging stage) was doubled (7.0 vs. 3.4 days) when a 0.25 wasp equivalent (WE) of sucrose gradient-purified McPDV was injected into a newly ecdysed fifth stadium host. Many of the treated larvae were unable to pupate, successfully and died at a point of incomplete larval-pupal ecdysis. Pupae that did result from the treated larvae weighed significantly less than controls, even at 0.025 WE. The rate of weight gain and extent of delay of development were dose-dependent; as little as 0.1 WE extended the time of active feeding by 1.5 days and yielded only 25% adults. A 0.05 WE dose yielded 78% adults compared to 95% for controls. The total protein content of hemolymph from individuals injected with McPDV was significantly less than that of controls at any McPDV dose equal to or greater than 0.1 WE. SDS-PAGE profiles of hemolymph proteins from control and McPDV-injected larvae revealed a marked inhibition of the normal accumulation of storage proteins during the fifth stadium and a lesser reduction of serine protease inhibitor protein. Thus, McPDV-injected larvae exhibited some symptoms (less total hemolymph protein and reduced amounts of storage protein) similar to those shown by both parasitized larvae and by larvae injected with M. croceipes teratocytes. However, McPDV affected development during the active feeding stage of the larva, while teratocytes primarily impacted larvae at the time when larval-pupal transformation processes are initiated. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 79-90 
    ISSN: 0739-4462
    Keywords: adipokinetic hormone ; cockroach ; cytochrome P450 ; fat body ; gene expression ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 103-111 
    ISSN: 0739-4462
    Keywords: Lepidoptera ; protease ; digestion ; midgut ; ectoperitrophic ; endoperitrophic ; intracellular ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The distribution of digestive proteinases in either the anterior and posterior midgut or between the midgut epithelium and ectoperitrophic and endo-peritrophic spaces in the midgut were examined in the European corn borer, Ostrinia nubilalis. Trypsin, chymotrypsin, elastase, and aminopeptidase activities were the same in the anterior and posterior halves of the midgut. Of the total aminopeptidase activity, 95% was located in the midgut epithelium, and 90% of the trypsin, 97% of chymotrypsin, and 93% of the elastase activity were found in the midgut lumen. Trypsin, measured by hydrolysis of benzoyl-L-arginine ethyl ester, and chymotrypsin levels were significantly higher in the ectoperitrophic space compared to the endoperitrophic space. Digestion in the midgut is proposed to be sequential with tryptic digestion occurring in the endoperitrophic space. Ingested protein is digested further in the ectoperitrophic space by the action of elastase, chymotrypsin, and a second trypsin. Final digestion occurs by an intracellular aminopeptidase. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 143-157 
    ISSN: 0739-4462
    Keywords: mosquito ; A. albopictus ; cultured cells ; transcription ; promoter ; RNA polymerase I ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the mosquito Aedes albopictus, two potential RNA polymerase I promoters that map 531 and 143 nucleotides upstream of the 18S rRNA gene have been defined on the basis of sequence homology with rRNA promoters from other species. Using the polymerase chain reaction, we confirmed that a 717 nucleotide region spanning the upstream (-531) and downstream (-143) promoters is homogeneous in genomic DNA and in cloned DNA. DNA probes representing each of these promoters, as well as upstream “spacer” promoters, exhibited protein-binding activity, and each unlabeled probe was an effective competitor of protein binding with the other probes, suggesting that these potential regulatory sequences interact with a common protein(s). Analysis of precursor ribosomal RNAs accumulated during temperature shock indicated that transcription is initiated primarily at the upstream (-531) promoter. RNAse protection and primer extension analyses confirmed the predominant use of this promoter, both in cultured cells and in mosquito life stages. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 207-207 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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  • 85
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    Archives of Insect Biochemistry and Physiology 28 (1995), S. 173-187 
    ISSN: 0739-4462
    Keywords: wing imaginal discs ; chitin synthesis ; Spodopfera frugperda ; ecdysteroid ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: 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.
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    Archives of Insect Biochemistry and Physiology 29 (1995) 
    ISSN: 0739-4462
    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 281-291 
    ISSN: 0739-4462
    Keywords: Lepidoptera ; haemolymph ; superoxide ; oxidative stress ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The superoxide generative system in Pseudaletia separata larval haemolymph plasma was shown to consist of at least two components, low (LMF) and high (HMF) molecular weight factors using ultrafiltration, dialysis, and gel filtration. The LMF was concluded to be a molecule(s) smaller than 5 kDa while the HMF to be a protein(s) larger than 100 kDa. The total amount of superoxide produced depended on the amount of LMF and was independent of that of HMF added to the reaction mixture. Both the LMF and HMF were required for O2- generation. From results in the present article, it was hypothesized that the LMF was a substrate(s) discharging electrons and HMF was an enzyme(s) to mediate the electron transfer to O2 forming O2-. Superoxide production was detected in the haemolymph plasma of 5 lepidopteran species in addition to P. separata. It was concluded that superoxide production is a common phenomenon at least in these lepidopterans. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 309-327 
    ISSN: 0739-4462
    Keywords: Lands cycle ; phospholipase A2 ; arachidonate ; phospholipids ; A23187 ; dopamine ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An important regulatory step for prostaglandin synthesis is the availability of the precursor, free arachidonic acid (AA). In isolated salivary glands of the lone star tick, Amblyomma americanum (L.), the level of free AA appears to depend on higher phospholipase A2 (PLA2) activity rather than decreased rates of re-esterification by lysophosphatide acyl transferase (LAT). This conclusion is supported by experiments where inhibition of LAT with merthiolate was without effect, while the calcium ionophore A23187, a PLA2 stimulant, increased levels of free AA. The PLA2 activity in A. americanum was reduced by the substrate analog, PLA2 inhibitor, oleyloxyethyl phosphorylcholine in a dose-dependent manner, but was insensitive to the other mammalian PLA2 inhibitors mepacrine (20μM), aristolochic acid (45μM), and dexamethasone (50μM). No substrate preference was observed for the functional group of the phospholipid, with phosphatidylcholine and phosphatidylethanolamine being equal sources of AA in A23187-stimulated glands. Compared to phospholipids containing other fatty acids, only arachidonyl-phospholipid (arachidonyl-PL) was significantly hydrolyzed by PLA2 activity in A23187-stimulated glands. Dopamine was as effective as A23187 as a stimulant of PLA2 activity in isolated glands, but this effect was abolished in the presence of the calcium channel blocking agent verapamil. It is concluded that free AA levels in tick salivary glands are increased through activation of a Type IV-like PLA2 following an increase of intracellular calcium caused by the opening of voltage-dependent calcium channels due to dopamine stimulation. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 397-414 
    ISSN: 0739-4462
    Keywords: adipokinetic hormone ; excitation ; flight behavior ; flight fuel mobilization and utilization ; migration, octopamine ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Male black cutworm, Agrotis ipsilon (Hufnagel) moths, were attached to laboratory flight mills on the third night after emergence. Individuals were classified according to their longest uninterrupted flight during the night. There were not enough non-fliers or trivial fliers (flight 〈 1 h) to make meaningful comparisons, so only short-fliers (flight ≥ 1 h but ≤ 5 h) and long-fliers (flights ≥ 5 h) were further tested. After a day of rest, short- and long-fliers were tethered to a stationary pin and forced to fly 0, 10, 20, 30, 60, 120, or 180 min, after which hemolymph was sampled and lipid and carbohydrate concentrations determined. Concentrations in samples taken at intervals after cessation of a 60-min flight indicated that both energy sources were being utilized during flight. Neck-ligation experiments indicated that a handling-induced hyperlipemic response occurs 15 min after the beginning of the sampling procedure, and that the hormone triggering does not come from the head. Resting concentration of lipid in long-fliers is about half that of short-fliers, but it increases threefold in the first 20 min of flight, whereas that of short-fliers remains unchanged. In-flight lipid concentration in controls, which experienced their first and only flight on day 5, was greater than the concentration in both short- and long-fliers. Between 10 and 20 min, carbohydrate concentrations decrease about twofold in long-fliers but drop only slightly in short-fliers. These data suggest that long-fliers may have a lower basal metabolic rate and depend more on carbohydrate when at rest than short-fliers and that long-fliers may utilize more carbohydrate than lipid during the initial period of flight. In addition, previous flight on the flight mills has a greater influence on lipid mobilization and usage in short-fliers than it does on long-fliers. Patterns of lipid mobilization during flight seem to differ between migrant and non-migrant species and to vary among migrants depending on the flight strategy used, but there are no major differences in patterns of carbohydrate usgaeg. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 41-59 
    ISSN: 0739-4462
    Keywords: PBAN-like substance ; Pheromone production ; Lepidoptera ; Noctuidae ; Incorporation ; Labeled precursors ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Pheromone production in the female turnip moth, Agrotis segetum, is under the control of a brain factor. This factor was demonstrated to be a proteinaceous substance termed pheromone biosynthesis activating neuropeptide-like substance (PBAN-like substance). The sex pheromone of Swedish A. segetum includes (Z)-5-decenyl acetate, (Z)-7-dodecenyl acetate, and (Z)-9-tetradecenyl acetate as major components. Decapitation of a female decreased pheromone production significantly. Pheromone production was restored by injection of homogenates of either male or female brain-suboesophageal ganglion or the corpora cardiaca alone. Pheromonotropic activity was also found in homogenates of the female thoracic ganglion and abdominal ganglion that were obtained during scotophase. Injection of female brain and thoracic ganglion homogenates made from insects during the scotophase induced two and four times as much Z7-12:OAc, respectively, as injection with similar homogenates from photophase. As little as one-eighth female equivalent (FE) brain homogenate was sufficient to increase the amount of Z7-12:OAc. The effect of brain homogenate on pheromone titer reached its maximum after 30 min. The activity of the PBAN-like substance present in female brain extracts was not correlated to the age of the donor. Injection of hemolymph collected during either photophase or scotophase into decapitated females did not increase the pheromone titer. The target site of the PBAN-like substance was not the pheromone gland, and the ventral nerve cord was not involved in the transportation of the PBAN-like substance, which implies a mode of action different from what has been reported in other moths. Brain homogenates obtained during photophase from females of African A. segetum, Spodoptera littoralis, or Ostrinia nubilalis as well as synthetic Bombyx-PBAN also induced pheromone production in decapitated Swedish female A. segetum. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 61-75 
    ISSN: 0739-4462
    Keywords: Cancer antennarius ; cholesterol ; crabs ; ecdysteroids ; high-density lipoproteins ; Y-organs ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cholesterol is the obligate precursor for ecdysteroid hormone synthesis by the ecdysial glands (Y-organs) in crustaceans, and all cholesterol in the hemolymph is bound to high-density lipoprotein (HDL). The mechanism was studied of how Y-organ cells acquire cholesterol. Y-organ segments were incubated with HDL isolated from hemolymph and labeled with 125I. After incubation, tissue was homogenized in acid to determine radioactivity in acid-precipitable (cell associated, intact) HDL and in acid-soluble (degraded) HDL. Both HDL uptake and degradation showed saturation kinetics. At saturation most of the total counts represented degraded HDL; by 3 h, degradation was 80%. Rates of HDL uptake and breakdown were higher in Y-organs from de-eyestalked crabs (deprived thereby of molt-inhibiting hormone, MIH) than in glands from intact crabs. Both parameters were depressed by inhibitors of glycolysis and oxidative phosphorylation dose dependently and by low temperature. HDL uptake also was depressed by cAMP added to the medium experimentally or through efflux from the tissue during incubation. These results indicate a mechanism for HDL uptake that entails receptor-mediated, energy-dependent endocytosis of the entire HDL-cholesterol complex. Also the results suggest that HDL uptake and degradation are mediated by cAMP and depressed by an eyestalk factor, presumably MIH. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 401-401 
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 351-367 
    ISSN: 0739-4462
    Keywords: Aphidiinae ; physiological interactions ; calyx fluid ; venom ; teratocytes ; in vitro rearing ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Biochemical and metabolic alterations in Acyrthosiphon pisum (Harris) (Homoptera: Aphididae) parasitized by the endophagous Aphidius ervi Haliday (Hymenoptera: Braconidae) are reported. Total ecdysteroid titer was determined by radioimmunoassay. The hemolymph levels of acylglycerols and proteins were assessed with an enzymatic method and Bradford's method, respectively. Alterations of the electrophoretic pattern of host hemolymph proteins were assessed by SDS-PAGE. Analyses were carried out on hosts parasitized as 1st instars, as well as on hosts which were parasitized later, as 3rd or 4th instars. Parasitism did not significantly alter the ecdysteroid titer of pre-adult stages when parasitized as 3rd instars. In contrast, the ecdysteroid titer in developmentally arrested 4th instars of A. pisum, previously parasitized as 1st instars, was significantly lower than in the case of synchronous nonparasitized controls. Ecdysteroids were detected also in adult aphids, which, when previously parasitized as 3rd instars, showed a precocious end of reproductive activity associated with a hormonal titer that was significantly lower than in actively reproducing nonparasitized controls of the same age. Acylglycerol and protein titers were significantly higher in the hemolymph of both early and late parasitized hosts, 5-6 days after parasitoid oviposition. SDS-PAGE analysis of hemolymph collected 5 days after parasitization revealed the presence of both upregulated proteins and of parasitoid-specific proteins. The observed biochemical changes in parasitized hosts were synchronized with the major part of parasitoid larval growth and, apparently, strictly related to parasitic castration of the host. The role and importance of host regulation factors controlling these biochemical and developmental alterations in parasitized pea aphids are discussed. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 93-94 
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    Keywords: Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 149-164 
    ISSN: 0739-4462
    Keywords: corpora allata ; adenylyl cyclase ; CAMP ; calcium ; calmodulin ; biogenic amines ; dopamine ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An assay was developed with which to study basic characteristics of an adenylyl cyclase in the corpora allata (CA) of the tobacco hornworm, Manduca sexta. The assay used glands collected and frozen at -80°C, to circumvent the problem of tissue availability. With this protocol for storage of tissue, less than 25% of the enzyme activity in fresh tissue was lost. Substances such as sodium fluoride (NaF) and Gpp(NH)p (a non-hydrolyzable GTP analog), which typically stimulate the adenylyl cyclases in other insect tissues, increased enzyme activity several-fold. There was a progressive decrease in the capacity of the CA adenylyl cyclase to be stimulated by NaF during the fifth stadium, suggesting a possible developmental change in the capacity of the associated G protein to be stimulated by NaF. The calcium/calmodulin (CaM) dependence of adenylyl cyclase activity was also investigated. The results demonstrated that addition of up to 10-4 M calcium to assays of enzyme activity in whole gland homogenates of both larval (day O) and prepupal (day 6) CA resulted in only a slight increase in the activity of the enzyme over basal rates in the presence of the calcium chelator EGTA. However, addition of as little as 5 m̈M CaM in the presence of 10-4 to 10-3 M calcium increased adenylyl cyclase activity three to five-fold. A similar stimulation was obtained with washed membrane preparations of day 0 and day 6 glands, but required a substantially higher concentration of CaM. Results demonstrated that the CA possess a calcium/CaM-dependent adenylyl cyclase from day 0 through day 6. A preliminary investigation of the effect of two biogenic amines on the CA adenylyl cyclase revealed that enzyme activity was not affected by octopamine, but a stage-specific effect was obtained with dopamine. Concentrations of 10-6 and 10-7 M stimulated enzyme activity in hornogenates of day 0 glands but inhibited activity in homogenates of day 6 CA. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 195-209 
    ISSN: 0739-4462
    Keywords: Drosophila ; apterous ; juvenile hormone ; germ-line clones ; interallelic complementation ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The apterous (ap) gene in Drosophila melanogaster encodes a homeodomain transcription factor. It is required for the development of the wings and of a subset of embryonic muscles. The gene has been implicated in the juvenile hormone (JH) system because mutations in ap lead to JH deficiency, and are associated with defective histolysis of the larval fat body, arrested vitellogenesis, sterility, and aberrant sexual behavior, all of which are dependent on JH.We describe here the use of hemizygotes and germ-line clones, of X-ray-and hybrid dysgenesis-induced lethal ap alleles to determine the primary role of the gene during development. We find that ap lethality is polyphasic, but occurs primarily at the larval and pupal stages. The lethal phenotype is not associated with any overt morphological abnormality, suggesting that death occurs from a systemic malfunction. Strong interallelic complementation for the wing phenotype was found between some ap mutations induced by hybrid-dysgenesis. By Northern blot analysis, we demonstrate an increase in ap expression in pupae and adults as compared to embryos and larvae, suggesting that it is developmentally regulated. Finally, primer extension is used to determine the transcription start site of the gene. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 239-254 
    ISSN: 0739-4462
    Keywords: prothoracicotropic hormone ; steroidogenesis ; cAMP ; calcium-dependent ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In both Manduca sexta and Drosophila melanogaster, metamorphic events are driven by ecdysteroids whose production in prothoracic gland (PGs) is stimulated periodically by neural factors. Differences in the life cycle of moths and flies have made it difficult to compare the regulation of ecdysteroid biosynthesis in these two species.As in Manduca, at least two neural factors in the larval Drosophila BVG complex were separable by molecular weight, and they stimulated increased ecdysteroid biosynthesis from the ring gland, a composite organ that includes PG cells. Drosophila neural extracts accelerated ecdysteroid biosynthesis in Manduca PGs and, conversely, partially purified Manduca PTTH preparations elevated ecdysteroid biosynthesis in Drosophila ring glands, suggesting that the two species may share structurally similar prothoracicotropic factors. Drosophila ring glands required the presence of calcium ions to respond to neural extracts, but the phosphodiesterase inhibitor MIX and cAMP analogues exerted little, if any, positive effect on production.Mean ecdysteroid production rates of BVG-ring gland complexes taken from Drosophila larvae during various phases of the wandering period were often submaximal and highly variable, suggesting that they fluctuate widely prior to pupariation. Based on available data in Drosophila and the Manduca model for the control of ecdysteroid biosynthesis, a developmental scheme for neuroendocrine control in Drosophila is proposed. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 271-293 
    ISSN: 0739-4462
    Keywords: ecdysteroids ; Manduca sexta ; prothoracicotropic hormone ; prothoracic glands ; second messengers ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The prothoracic glands of the tobacco hornworm, Manduca sexta, respond to prothoracicotropic hormone (PTTH) by a regulatory pathway involving cAMP, protein phosphorylation, protein synthesis, and enhanced secretion of ecdysteroids including ecdysone and 3-dehydroecdysone. Recent investigations have revealed that PTTH acts by this general mechanism throughout the fifth larval instar, i.e., during the transition from larva to pupa. However, the glands undergo developmental changes in size, steroidogenic capacity, and in elements of the signalling pathway associated with synthesis, degradation, and intracellular action of cAMP. The present review describes such changes, and their possible regulation and consequences, in the general context of endocrine events underlying larval-pupal metamorphosis during the fifth larval stage. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 30 (1995), S. 225-237 
    ISSN: 0739-4462
    Keywords: Drosophila melanogaster ; corpus allatum ; juvenile hormone ; radiochemical assay ; allatostatic factor ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Adult female Drosophila melanogaster corpus allatum (CA) synthesize JHB3 from endogenous and exogenous precursors in vitro. We present evidence supporting the thesis that biosynthesis proceeds from precursor FA via initial epoxidation and terminal methylation on the basis of the following: (1) Methyl farnesoate is not epoxidized to JHIII or JHB3; (2) Authentic JHIII is not epoxidized to JHB3; and (3) FABE is markedly metabolized to JHB3. Cerebral allatostatic factors act at some stage subsequent to FA and this precursor is not normally rate-limiting. Additionally, neural inhibition from the brain acts at some biosynthetic step prior to FA. © 1995 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 30 (1995), S. 295-306 
    ISSN: 0739-4462
    Keywords: radioimmunoassay ; JH I ; JH II ; JH III ; hemolymph ; insect hormone ; Manducasexta sexta ; Hyalophora cecropia ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recent refinements in juvenile hormone radioimmunoassay technology now make this method significantly more sensitive and easier to use. Rabbit poly-clonal antisera against (10R) JH III and racemic JH II have been developed to determine hemolymph hormone titers in the low picogram range. The antisera display minimal cross-reactivity with JH metabolites, JH analogs, and hemolymph lipids. One antiserum recognizes racemic JH I, II, and (10R) III almost equivalently, exhibiting 50% displacement between 100 and 130 pg per tube. Another antiserum is JH II-specific and exhibits 50% displacement at 35 pg per tube. Assay sensitivity has been enhanced by using (10R,11S) [methyl-3H]-JH II of very high specific activity (〉 80 Ci/mmol) generated with Hyalophora cecropia accessory gland S-adenosylmethionine transferase and S-[methyl-3H]-adenosyl-L-methionine. Preparation of biological samples has been simplified with overall recoveries of JH from hemolymph ranging between 60 and 75%. © 1995 Wiley-Liss, Inc.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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