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At least 163 records · Page 9Linked to original sources
PubMed1984

The glutathione status of Ephestia cautella (Walker) pupae exposed to carbon dioxide.

The exposure of E. cautella pupae, 0-24 hr old, to controlled atmospheres high in carbon dioxide reduces their tissue glutathione levels. If the period of exposure is not too long the levels return to control values after three days. Evidence is presented to show that exposure to CO2 inhibits the biosynthesis of glutathione. The implications of such inhibition at cellular and higher levels are discussed.

Animals↗
PubMed1995

Neuromuscular development in Drosophila: insights from embryos and pupae.

During embryogenesis, Drosophila melanogaster generates a set of muscles and motoneurons that are restructured during pupal metamorphosis to give rise to an adult motor system. The study of neuromuscular development in embryos and pupae has recently revealed important insights into the cellular and molecular mechanisms controlling both muscle patterning and the establishment of synaptic connections.

Animals↗
PubMed2005

Efficient large-scale protein production of larvae and pupae of silkworm by Bombyx mori nuclear polyhedrosis virus bacmid system.

Silkworm is one of the most attractive hosts for large-scale production of eukaryotic proteins as well as recombinant baculoviruses for gene transfer to mammalian cells. The bacmid system of Autographa californica nuclear polyhedrosis virus (AcNPV) has already been established and widely used. However, the AcNPV does not have a potential to infect silkworm. We developed the first practical Bombyx mori nuclear polyhedrosis virus bacmid system directly applicable for the protein expression of silkworm. By using this system, the green fluorescence protein was successfully expressed in silkworm larvae and pupae not only by infection of its recombinant virus but also by direct injection of its bacmid DNA. This method provides the rapid protein production in silkworm as long as 10 days, is free from biohazard, thus will be a powerful tool for the future production factory of recombinant eukaryotic proteins and baculoviruses.

Animals↗
PubMed2005

Isolation of mannose-binding C-type lectin from Heliothis virescens pupae.

A mannose-binding C-type lectin (MBL) was isolated by affinity chromatography from Heliothis virescens immune pupal hemolymph. The immune pupal hemolymph was obtained after bacterial injection of live Enterobacter cloacae bacteria. MBL in mammals acts as an opsonin for phagocytosis and activates the lectin complement pathway of the innate immune response, which leads to killing of gram-negative bacteria and enveloped viruses. The affinity-purified and reduced pupal MBL showed a single band of 36 kDa by SDS-PAGE (12% gel). A dot-immunoblot ELISA (using guinea pig anti-MBL IgG as primary antibody) demonstrated specificity of the antibody for the affinity-purified pupal MBL. The immune pupal hemolymph contained 21 microg of MBL per ml of hemolymph. The amino acid composition of the purified pupal MBL was determined with high amounts of arginine and histidine detected. The presence of MBL in insect pupae has not before been reported and could be important in pupal innate immunity to bacterial infection.

Amino Acid Sequence↗
PubMed2006

Safety evaluation of protein of silkworm (Antheraea pernyi) pupae.

The protein of silkworm pupae (PSP) has been thought to be a new available source of high quality protein that contains all the amino acids needed by the human body. The safety of PSP was evaluated systematically by a series of acute and sub-acute toxicological tests: (i) Acute toxicity test: The oral maximum tolerated dose of PSP was more than 15.0 g/kg body weight in mice, due to the absence of toxicity according to the criteria of acute toxic classifications; (ii) Mutagenicity test: PSP had no mutagenicity, as judged by a negative Ames test, mouse bone marrow cell micronucleus test and mouse sperm abnormality test; (iii) 30 days feeding study: No deaths or abnormal hematological, clinical chemical and histopathological changes and clinical signs had been found in rats when administrated PSP at 0.30, 0.75 and 1.50 g/kg/day to the rats for 30 days in each group during the test, respectively. No statistically significant differences had been found in body weights, food consumption and food efficiency of rats in each test group (P>0.05). These results indicate that PSP can be generally regarded as safe at a maximum dose of 1.50 g/kg/day in rats.

Animals↗
PubMed2003

The Drosophila SSL gene is expressed in larvae, pupae, and adults, exhibits sexual dimorphism, and mimics properties of the beta subunit of casein kinase II.

Drosophila melanogaster casein kinase II (CKII) is composed of catalytic alpha and regulatory beta subunits that generate the alpha2beta2 holoenzyme. A two-hybrid screen of a Drosophila embryo library using CKIIalpha as bait has resulted in the isolation of multiple cDNAs encoding SSL, a CKIIbeta-like polypeptide. We demonstrate that CKIIbeta, beta', and SSL exhibit robust and comparable interaction with CKIIalpha. Residues in SSL that mediate interaction with CKIIalpha appear similar to those in CKIIbeta, and SSL forms homodimers and heterodimers with CKIIbeta or beta' as well. We have tested all known Drosophila CKIIbeta-like proteins for rescue of the ion-homeostasis defect of yeast lacking beta subunits and find that CKIIbeta and SSL complement, beta' has marginal function, and Stellate appears non-functional. We have used real-time RT-PCR to assess developmental expression, and find that CKIIbeta is robust and ubiquitous, whereas SSL is restricted to males (third-instar-larvae, pupae, and adults), but is nondetectable in females of the corresponding stages. These results indicate that SSL expression encompasses a greater developmental window than that previously suggested and may confer distinct functions to CKII in a sex-specific manner.

Amino Acid Sequence↗
PubMed2003

Mechanical aspects of heartbeat reversal in pupae of Manduca sexta.

Pulsations of the dorsal vessel were investigated with new optocardiographic techniques based on the transmission and reflection of pulse-light through optic fibers. This noninvasive technique enabled simultaneous, in vivo multisensor recordings of the heartbeat without touching the pupal integument. There was a very regular heartbeat reversal with 3 distinctive phases: (a) a backward-oriented (retrograde) cardiac pulsation; (b) a forward-oriented (anterograde) pulsation with faster frequency; and (c) shorter or longer periods of temporary cardiac standstill that usually occurred after the termination of the anterograde phase. Occasionally, there were localized series of systolic cardiac contractions during the retrograde phase. Simultaneous recordings from the base and the tail of the abdomen revealed a reciprocal, "mirror image-like", quantitative relationship. The most intensive anterograde hemolymph flow occurred at the base while the most intensive retrograde flow occurred at the tail of the abdomen. The bi-directional switchovers of heartbeat (reversal) were occasionally associated with modifications during each of the unidirectional cardiac phases. Anterograde peristalsis showed a 2-fold higher frequency of pulsation in the thoracic aorta in comparison with the posterior parts of the heart. Thus, in addition to the "odd" peristaltic waves originating at the tail, there were intercallated "even" peristaltic waves originating in the middle of the abdomen. Both of them propagated hemolymph through the thoracic aorta into the head; the first waves took the hemolymph in from the distal end, while the second sucked it from the middle of the abdomen. The use of multiple optocardiographic sensors also enabled detection of cardiac pulsations on the opposite, ventral side of the body, within the ventral perineural sinus. The ventral side of the head showed only the presence of an anterograde pulse, whereas the ventral side of the tail exhibited a strong reciprocal retrograde phase and a very weak anterograde phase. These results explain why the existence of a periodic heartbeat reversal should be essential for circulatory functions at both extremities of the cylindrical insect body. In diapausing pupae, regular cycles of heartbeat reversal were substituted by prolonged periods of anterograde pulsation during the entire duration of bursts of CO2 release (average duration of the burst was 18-20 min, periodicity 5 to 18 h). The physiological nature of such feed-back correlation between heartbeat and metabolic CO2 production is not yet clear, because the anterograde heartbeat could be also induced by a number of nonspecific factors unrelated to CO2 (mechanical irritation, injury, injections, elevated temperature). During the postdiapause, developing pharate-adult stage, the correlation between CO2 and anterograde heartbeat completely disappeared. It has been concluded that regulation of insect heartbeat represents a highly coordinated, myogenic stereotype with inherent rhythmicity, which can be modified by a number of external and internal factors.

Animals↗
PubMed2000

Structure of insect chitin isolated from beetle larva cuticle and silkworm (Bombyx mori) pupa exuvia.

Chitin samples in a alpha-form structure were isolated from beetle larva cuticle and silkworm (Bombyx mori) pupa exuvia by treatment with 1 N HCl and 1 N NaOH. Chitosan was prepared by treating them in 40% NaOH containing NaBH(4). Chitin and chitosan were analyzed by X-ray, [13C]CP/MAS NMR, [13C]FT-NMR, and scanning electron microscopy (SEM) methods. Insect chitin degraded more readily than shrimp chitin when treated with 6 N HCl and the enzyme-chitinase. After treatment with 2 N HCl at 100 degrees C, the insect chitin crystallinity increased. N-deacetylation of insect chitin was easier than that of crustaceous chitin, and about 94% of the N-acetyl groups were removed in one treatment with 40% NaOH for 4 h at 110 degrees C. After treatment with 2 N HCl, 55% of the N-acetyl groups of silkworm chitin were removed under the same conditions. Beetle chitin showed a higher affinity for chitinase than shrimp chitin.

Animals↗
PubMed2003

Utilization of fermented silkworm pupae silage in feed for carps.

Fermented silkworm pupae (SWP) silage or untreated fresh SWP pastes were incorporated in carp feed formulations replacing fishmeal. The feed formulations were isonitrogenous (30.2-30.9% protein) and isocaloric (ME = 2905-2935 kcal/kg). Feeding under a polyculture system consisting of 30% each of catla (Catla catla), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita) with 10% silver carps (Hypophthalmychthys molitrix) was carried out in ponds to evaluate the nutritive quality of SWP silage. Survival rate, feed conversion ratio and specific growth rate, respectively, were 84.2%, 2.10 and 2.39 for fermented SWP silage, 65.8%, 2.98 and 2.26 for untreated SWP and 67.5%, 3.16 and 2.20 for fishmeal indicating clearly that the fermented SWP silage was nutritionally superior to untreated SWP or fishmeal. The dietary influence on the proximate composition of whole fish was marginal.

Animals↗
PubMed2002

Purification and characterization of apolipophorin III from immune hemolymph of Heliothis virescens pupae.

Apolipophorin III (ApoLp-III) from Heliothis virescens pupae was purified by heat-treatment followed by Sephadex G-50 filtration and reverse phase-HPLC. The molecular mass of the purified ApoLp-III was determined as 17965.9+/-5 Da by mass spectrometry. The N-terminal sequence confirmed the protein as ApoLp-III with homology of 56-83% to other insect ApoLp-III molecules. The amino acid spatial arrangement of the predicted alpha-helix 1 of Heliothis ApoLp-III was nearly identical to that of the amphipatic alpha-helix 1 of Manduca sexta ApoLp-III. The absorption spectrum from 240-340 nm of the Heliothis ApoLp-III was the same as the UV spectra of ApoLp-III from Manduca sexta and Galleria mellonella, showing absorption maxima at 280, 268, 264 and 259 nm. These results indicated that the primary structure of ApoLp-III is conserved in lepidopterans. The Heliothis ApoLp-III was not a glycoprotein and showed hemagglutination activity against rabbit red blood cells. This hemagglutination activity was abolished by Tween 80, but not by six different carbohydrates. Hydrophobic interaction of ApoLp-III with red blood cells agreed with structural studies since ApoLp-III binds lipid through hydrophobic interaction after conformational change. Bacterial injection apparently increased the amount of ApoLp-III in immune hemolymph when compared with normal hemolymph, and may indicate that ApoLp-III plays a role in insect immunity.

Amino Acid Sequence↗
PubMed2004

Enzymatic characterization and functional groups of polyphenol oxidase from the pupae of blowfly (Sarcophaga bullata).

Polyphenol oxidase (EC 1.14.18.1) was purified from the pupae of blowfly (Sarcophaga bullata) by a procedure involving ammonium sulfate fractionation and chromatography on DEAE-cellulose and Sephadex G-100. Kinetic characteristics of the enzyme were determined using L-DOPA as substrate. The specific activity of the enzyme was 770 U/mg, and the Michaelis constant (Km) was 1.5 +/- 0.1 mM (pH 6.8, 30 degrees C). Activity was maximal at 40 degrees C, pH 6.5. Chemical modification experiments demonstrated that cysteine and tryptophan residues are essential and arginine residues are not essential to the enzyme function. The enzyme is inhibited by quercetin with an IC50 of 0.20 +/- 0.06 mM. The inhibition is of competitive type, and the inhibition constant was determined to be 88 micro M.

Animals↗
PubMed2000

Viresin. A novel antibacterial protein from immune hemolymph of Heliothis virescens pupae.

Immune hemolymph was collected from fifth instar larvae and 1-day-old pupae of Heliothis virescens after injection of prepupae with live Enterobacter cloacae. Induction of antibacterial activity against Escherichia coli K12 D31 was 7.5 times greater in pupal than in larval immune hemolymph. Lysozyme activity of immune pupal hemolymph against Micrococcus lysodeikticus was 11 times greater when compared with lysozyme activity of immune larval hemolymph. Early pupal immune response with regard to antibacterial activity was much greater than larval immune response in H. virescens. Normal pupal hemolymph showed an increase in antibacterial activity and lysozyme that was induced during metamorphosis. Antibacterial protein was isolated together with lysozyme by gel filtration chromatography and then separated from lysozyme by sequential electrophoresis with a native acid gel and SDS gel. Molecular mass of antibacterial protein was estimated to be 12 kDa. The N-terminal amino acid sequence of 12-kDa protein was different from those of antibacterial molecules found in other insects and has not been identified before. A sample containing 12-kDa protein was negative for immunoblotting with anti-synthetic cecropin B antibody. We have named the novel 12-kDa antibacterial protein viresin. Viresin showed antibacterial activity against several Gram-negative bacteria including E. cloacae but not against Gram-positive bacteria.

Amino Acid Sequence↗
PubMed1962

Temperature effects on day old Drosophila pupae.

Day old Drosophila pupae were subjected to a variety of closely controlled temperature shocks. Twenty-five hours after puparium formation (at 23 degrees ), temperatures from 39.5-41.5 degrees (Q(1) = 2.3) differentially disturb the formation of the posterior crossvein. Three other separate treatments disturb posterior crossvein formation: treatments in the range 36.0-37.0 degrees at 25 hours; 37.3-37.8 degrees at 25 hours; and 39.5-41.5 degrees at 19 hours. Certain qualitative effects are associated with certain temperatures: elliptical holes are seen in wings of flies exposed 25 hours after puparium formation to temperatures from 37.3-37.8 degrees . Anterior crossvein defects ensue if animals are similarly exposed to temperatures from 37.9-38.2 degrees . Within the physiological range, animals raised at higher temperatures are more resistant to the effects of temperatures at 39.5-41.5 degrees . An extremely rapid temperature adaptation by exposures to temperatures in the range 31-38 degrees results in markedly greater resistance to heat shock; here resistance to production of crossvein defects increases faster than to death. The association between qualitative effects and treatment temperatures is modified by changing the temperature at which the animals spend their first day of pupal life. Summation experiments support conclusions drawn from the simpler experiments. Genetic variation and interspecific variation are discussed in the present context, as well as implications of the role of protein denaturation in the biological effects of high temperatures and further, more general experiments.

Adaptation, Physiological↗
PubMed1993

Purification and characterization of prophenoloxidases from pupae of Drosophila melanogaster.

Two isoforms of prophenoloxidase were isolated from pupae of Oregon-R strain of Drosophila melanogaster. The purification procedure included ammonium sulfate fractionation, Sephacryl S-200 gel chromatography, DEAE-cellulose, and hydroxylapatite column chromatography. The two isoforms, A1 and A3, could be separated by ammonium sulfate fractionation. The isoelectric points of A1 and A3 were determined to be pH 5.8 and 6.7, respectively. The molecular weights of the monomers of A1 and A3 were estimated by SDS-PAGE to be 78 and 77 kDa, respectively. The native states of A1 and A3 are considered to be homodimeric, as judged by gel-filtration chromatography.

Ammonium Sulfate↗
PubMedDate not supplied

Green pupa (gp), a male lethal in culex quinquefasciatus.

An autosomal recessive mutant, green pupa (gp), in Culex quinquefasciatus has been isolated. The homozygous (gp/gp) females are viable whereas male homozygotes do not develop beyond the pupal stage. The behavior of this mutant character in pedigree studies has been reported.

Animals↗
PubMed1980

Insect immunity. Purification and properties of three inducible bactericidal proteins from hemolymph of immunized pupae of Hyalophora cecropia.

Three inducible bacteriolytic proteins, designated P7, P9A and P9B, from the hemolymph of immunized pupae of the giant silk moth Hyalophora cecropia have been purified using a two-step procedure with cation-exchange chromatography. Purified protein P7 has a molecular weight of 15000 and its amino acid composition shows a great similarity to that of the lysozyme from the wax moth Galleria mellonella. Moreover, heat stability, pH-rate profile and bacteriolytic specificity also indicate that protein P7 is a lysozyme. The other purified proteins, P9A and P9B, are highly potent against Escherichia coli and some other gram-negative bacteria. The amino acid compositions of proteins P9A and P9B are very similar, although the contents of glutamic acid and methionine were different. The molecular weights of these very basic proteins are around 7000. The P9 proteins are heat stable; their activities were retained after 30 min incubation at 100 degrees C. Both forms of protein P9 clearly differ from the lysozyme class of enzymes and they may represent a new type of bacteriolytic protein.

Amino Acids↗
PubMed1985

Glycosphingolipids in insects. Chemical structures of ceramide monosaccharide, disaccharide, and trisaccharide from pupae of Calliphora vicina (Insecta: Diptera).

The presence of glycosphingolipids in the pupae of the blowfly, Calliphora vicina, was established. The thin layer chromatographic pattern of the total neutral glycolipids revealed the presence of more than 13 components, the major one being ceramide monohexoside. By the use of high performance liquid chromatography, the three simplest components were isolated and their chemical structures determined: Glc(beta 1-1)Cer, Man(beta 1-4)-Glc(beta 1-1)Cer [with minor component Gal(beta 1-4)Glc(beta 1-1)Cer] and GlcNAc(beta 1-3)Man(beta 1-4)Glc(beta 1-1)-Cer. The ceramide composition of the parent insect glycosphingolipids is dominated by the 20:0 fatty acid, arachidic acid, and the sphingoid tetradecasphing-4-enine.

Animals↗
PubMed1964

GIANT POLYTENE CHROMOSOMES IN HYPODERMAL CELLS OF DEVELOPING FOOTPADS OF DIPTERAN PUPAE.

Large hypodermal cells with giant polytene chromosomes are present within the developing feet of Sarcophaga and other fly pupae. The cells secrete a large volume of cuticle; at this time there is extensive "puffing" of the chromosomes. Being hypodermal and superficially situated with permanent secretory products, the cells could be ideal experimental and genetical tools.

Anatomy↗