bluuebunny/crossref_metadata_2025 huggingface dataset:
Unusual variants of Alexander's disease
Authors: van der Knaap Marjo S. , Salomons Gajja S. , Li Rong , Franzoni Emilio , Gutiérrez‐Solana Luiz González , Smit Leo M. E. , Robinson Richard , Ferrie Collin D. , Cree Bruce , Reddy Alyssa , Thomas Neil , Banwell Brenda , Barkhof Frederik , Jakobs Cornelis , Johnson Anne , Messing Albee , Brenner Michael
Published: February 2005

The purpose of this study was to describe unusual variants of Alexander's disease. We studied 10 patients who did not meet previously established magnetic resonance imaging (MRI) criteria for Alexander's disease, but for whom this diagnosis was considered because of Rosenthal fibers at histological examination or presence of some MRI features suggestive of Alexander's disease. Sequence analysis of theGFAPgene was performed. In eight patients, MRI results showed predominantly posterior fossa lesions, especially multiple tumor‐like brainstem lesions. One patient had asymmetrical frontal white matter abnormalities and basal ganglia abnormalities. One patient (Patient 10) developed degeneration of the frontal white matter. In nine patients, a mutation was found that was concluded to be pathogenic, because the mutation was de novo (five patients), a known mutation was found (two patients), or assembly of the glial fibrillary acidic protein was abnormal in cultured cells (two patients). In Patient 10, a DNA variation was found that was also present in the patient's clinically unaffected father and was concluded to be a polymorphism. In conclusion, DNA diagnostics is warranted in patients who display MRI features suggestive of Alexander's disease, even if they do not meet the full set of previously established MRI criteria. Ann Neurol 2005;57:327–338
Cerebral fat embolism: Usefulness of magnetic resonance spectroscopy
Authors: Guillevin Rémy , Vallée Jean N. , Demeret Sophie , Sonneville Romain , Bolgert Francis , Mont'Alverne Francisco , Deseilligny Charles Pierrot , Chiras Jacques
Published: February 2005

We report a case of cerebral fat embolism which occurred in a 33‐year‐old man after a diaphyseal femoral fracture without cranial traumatism. The initial examination showed an incomplete picture of coma with tetrapyramidal syndrome and cutaneomucous purpura. There was no respiratory damage. We present a magnetic resonance spectroscopy analysis of the cerebral lesions observed in the initial phase of the embolism, as well as follow‐up, which has strengthened the clinical and imaging features for the diagnosis. Ann Neurol 2005;57:434–439
Familial Parkinson's disease: Clinical and genetic analysis of four Basque families
Authors: Paisàn‐Ruìz Coro , Sàenz Amets , de Munain Adolfo Lòpez , Martì Itxaso , Martìnez Gil Angel , Martì‐Massò Josè F. , Pèrez‐Tur Jordi
Published: February 2005

During the last few years several loci have been linked to Mendelian forms of Parkinson's disease (PD). To date, 5 causative genes of 10 identified loci are known, and they have provided enormous insight into the molecular pathways involved in this common neurodegenerative disorder. One of the recently identified loci, PARK8, causes autosomal dominant PD with, apparently, various degrees of abnormal deposition of α‐synuclein or τ in the neuronal cells in the pedigrees currently reported. We genetically characterized four Basque families and found evidence for linkage of autosomal dominant PD to the PARK8 locus, with a maximum 2‐point logarithm of odds score of 3.21 (θ = 0.00) for marker D12S345. The clinical features of these families are those of typical PD, including good response to levodopa therapy, rigidity, and akinesia, and a mean age of 55 years at disease onset. Ann Neurol 2005;57:365–372
A mutant PSEN1 causes dementia with lewy bodies and variant Alzheimer's disease
Authors: Ishikawa Atsushi , Piao Yue‐Shan , Miyashita Akinori , Kuwano Ryozo , Onodera Osamu , Ohtake Hiroaki , Suzuki Masahiro , Nishizawa Masatoyo , Takahashi Hitoshi
Published: February 2005

We report early‐onset parkinsonism and dementia of 18 years' duration in a 52‐year‐old man whose grandfather and father had suffered from a similar neurological disease. In this patient, we found neuronal loss in various brain regions including the substantia nigra and cerebral cortex, Lewy bodies, cotton wool plaques, corticospinal tract degeneration, cerebral amyloid angiopathy, and a novel three–base pair deletion in exon 12 of the presenilin‐1 (PSEN1) gene. We considered that the mutant PSEN1 might play an important role in the pathogenetic process of both aggregation of α‐synuclein into Lewy bodies and deposition of β‐amyloid into cotton wool plaques. Ann Neurol 2005;57:429–434
Sensorimotor returning in complex regional pain syndrome parallels pain reduction
Authors: Pleger Burkhard , Tegenthoff Martin , Ragert Patrick , Förster Ann‐Freya , Dinse Hubert R. , Schwenkreis Peter , Nicolas Volkmar , Maier Christoph
Published: February 2005

Patients with complex regional pain syndrome (CRPS) and intractable pain showed a shrinkage of cortical maps on primary (SI) and secondary somatosensory cortex (SII) contralateral to the affected limb. This was paralleled by an impairment of the two‐point discrimination thresholds. Behavioral treatment over 1 to 6 months consisting of graded sensorimotor retuning led to a persistent decrease in pain intensity, which was accompanied by a restoration of the impaired tactile discrimination and regaining of cortical map size in contralateral SI and SII. This suggests that the reversal of tactile impairment and cortical reorganization in CRPS is associated with a decrease in pain. Ann Neurol 2005;57:425–429
Changes in the neuromuscular synapse induced by an antibody against gangliosides
Authors: Santafé Manel M. , Sabaté M. Mar , Garcia Neus , Ortiz Nico , Lanuza M. Angel , Tomas Josep
Published: February 2005

In this study, we used a monoclonal IgM antibody from a patient with a pure motor chronic demyelinating polyneuropathy, which binds specifically to the complex gangliosides GM2, GalNAc‐GD1a, and GalNAc‐GM1b, which appear to have a common epitope of ‐[GalNAcβ1‐4Gal(3‐2αNeuAc)β1]. This was done for the following reasons: (1) to localize these gangliosides in specific cellular components of the neuromuscular junction (NMJ), and (2) to describe the anti–ganglioside antibody–induced structural and functional changes in the NMJs to gain insight into the role of gangliosides in the synaptic function. Using immunofluorescence techniques, we found that these gangliosides are located only in the presynaptic component of the motor end‐plates, both in nerve terminals and in Schwann cells. After 2 weeks of continued passive transfer of the IgM monoclonal antibody over the mouse levator auris longus muscle, electromyography showed an axonal or NMJ disorder. Morphology showed important nerve terminal growth and retraction changes. Using intracellular recording electrophysiology, we found neurotransmitter release alterations, including quantal content reduction and an immature expression of voltage‐dependent calcium channels similar to what occurred during NMJ development and regeneration. These changes were complement independent. The results showed that these gangliosides were involved in the reciprocal Schwann cell–nerve terminal interactions, including structural stability and neurotransmission. Ann Neurol 2005;57:396–407
Glial fibrillary acidic protein mutations in infantile, juvenile, and adult forms of Alexander disease
Authors: Li Rong , Johnson Anne B. , Salomons Gajja , Goldman James E. , Naidu Sakkubai , Quinlan Roy , Cree Bruce , Ruyle Stephanie Z. , Banwell Brenda , D'Hooghe Marc , Siebert Joseph R. , Rolf Cristin M. , Cox Helen , Reddy Alyssa , Gutiérrez‐Solana Luis González , Collins Amanda , Weller Roy O. , Messing Albee , van der Knaap Marjo S. , Brenner Michael
Published: February 2005

Alexander disease is a progressive, usually fatal neurological disorder defined by the widespread and abundant presence in astrocytes of protein aggregates called Rosenthal fibers. The disease most often occurs in infants younger than 2 years and has been labeled a leukodystrophy because of an accompanying severe myelin deficit in the frontal lobes. Later onset forms have also been recognized based on the presence of abundant Rosenthal fibers. In these cases, clinical signs and pathology can be quite different from the infantile form, raising the question whether they share the same underlying cause. Recently, we and others have found pathogenic, de novo missense mutations in the glial fibrillary acidic protein gene in most infantile patients examined and in a few later onset patients. To obtain further information about the role of glial fibrillary acidic protein mutations in Alexander disease, we analyzed 41 new patients and another 3 previously described clinically, including 18 later onset patients. Our results show that dominant missense glial fibrillary acidic protein mutations account for nearly all forms of this disorder. They also significantly expand the catalog of responsible mutations, verify the value of magnetic resonance imaging diagnosis, indicate an unexpected male predominance for the juvenile form, and provide insights into phenotype–genotype relations. Ann Neurol 2005;57:310–326
Mutant valosin‐containing protein causes a novel type of frontotemporal dementia
Authors: Schröder Rolf , Watts Giles D. J. , Mehta Sarju G. , Evert Bernd O. , Broich Petra , Fließbach Klaus , Pauls Katharina , Hans Volkmar H. , Kimonis Virginia , Thal Dietmar R.
Published: February 2005

Mutations in the valosin‐containing protein (VCP) gene on chromosome 9p13‐p12 recently have been shown to cause autosomal dominant inclusion body myopathy associated with Paget's disease of the bone and frontotemporal dementia. Here, we report the central nervous system autopsy findings in a 55‐year‐old German patient with inclusion body myopathy and frontotemporal dementia who harbors a heterozygous R155C missense mutation residing in the N‐terminal CDC48 domain of VCP, which is involved in ubiquitin binding. We demonstrate that mutant VCP causes a novel type of frontotemporal dementia characterized by neuronal nuclear inclusions containing ubiquitin and VCP. Ann Neurol 2005;57:457–461
Mechanical properties and structure of high‐density polyethylene samples prepared by injection molding with low‐frequency vibration
Authors: Zhang Jie , Shen Kaizhi , Gao Yuegin , Yuan Yi
Published: February 2005

To better understand the formation of different crystal structures and improve the mechanical properties of high‐density polyethylene samples, melt vibration technology, which generally includes shear vibration and hydrostatic pressure vibration, was used to prepare injection samples. Through melt vibration, the crystal structure changed from typical spherulites of the traditional injection sample to obviously orientated lamellae of vibration samples. Sizes and orientation degrees of lamellae were different according to different vibration conditions. Crystallinity degrees of vibration samples increased notably. Therefore, the tensile strength of vibration samples increased with increasing vibration frequency and vibration pressure, whereas elongation of vibration samples decreased during the first stage and then continued to increase as the vibration frequency increased. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 96: 818–823, 2005
Double‐shell surface modifications of large poly(methyl methacrylate) spheres
Authors: Guo Weihong , Durant Yvon G. , Wang Jikui , Wu Chifei
Published: February 2005

Two reactive shells were polymerized on poly(methyl methacrylate) (PMMA) spheres to improve the compatibility of the spheres with a paint binder. By adjusting the buoyancy of the polymerization medium, PMMA, retroreflective spheres with an average diameter of 600 μm were successfully modified with a thin cross‐linked layer together with an amine functional layer outside on the surface. The cross‐linked layer was polymerized by methyl methacrylate and ethylene glycol dimethacrylate by suspension polymerization. After that, a second monomer with functional amine group was polymerized and a functional layer was formed on the surface. The amine groups on the modified spheres can react with the enamine groups of acetoacetate already formulated in the binder and connected the spheres and the paint by chemical bond. The structure of the modified spheres was studied using transmission election microscopy, FTIR, DSC, and other tests. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 96: 829–836, 2005
Radiation synthesis of copolymers for adsorption of dyes from their industrial wastes
Authors: Abd El‐Aal Samia E. , Hegazy El‐Sayed A. , AbuTaleb M. F. , Dessouki A. M.
Published: February 2005

Copolymer hydrogels were prepared by γ‐radiation copolymerization of N‐vinyl‐2‐pyrrolidone (NVP) and methylmethacrylate (MMA) or acrylonitrile (AN). The effect of NVP/MMA and NVP/AN compositions and irradiation dose on the gel fraction yield in the prepared hydrogels was determined. It was found that as the content of NVP increased, the gel fraction yield decreased. The increase in irradiation dose resulted in increasing the crosslinked network structure and, consequently, the gel percentage increased. The thermal stability and swelling properties were also investigated as a function of comonomer composition. The nitrile groups (–CN) in the prepared copolymer were converted into their respective amidoxime groups by treating with NH2OH‐hydrochloride. Such a process resulted in improving both the swelling behavior and adsorption capacity. Characterization and some selected properties of the prepared hydrogels were studied and accordingly the possibility of its practicable use in the treatment of industrial waste dyes was determined. The thermodynamic parameters for the adsorption process were calculated. The effect of treatment time, pH of feed solution, effect of initial feed concentration, and temperature on the dye uptake was also investigated. The maximum uptake for investigated dyes was higher for treated NVP/AN hydrogel than that for untreated NVP/AN and NVP/MMA ones. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 96: 753–763, 2005
Comparison of sterols of pollens, honeybee workers, and prepupae from field sites
Authors: Svoboda James A. , Herbert Elton W. , Lusby William R. , Thompson Malcolm J.
Published: February 2005

Sterols from pollen collected by foraging honeybees, Apis mellifera L, at seven field sites were compared with the sterols of foraging adults and/or prepupae collected from colonies at each site. Invariably, the composition of prepupal sterols was comparable to that found in previous cage studies using chemically defined diets containing various dietary sterols: 24‐methyl‐enecholesterol was the major sterol; sitosterol and isofucosterol were present in lesser, but significant amounts; and a trace amount of cholesterol was identified in each sample. This occurred even though some of the pollen sterols contained little 24‐methylenecholesterol, sitosterol, or isofucosterol and a preponderance of certain other sterols, such as δ7‐stigmasten‐3β‐ol and δ7,24(28)‐campestadien‐3β‐ol in goldenrod and corn pollens, respectively. Thus the selective transfer and utilization of sterols in honeybees that have been demonstrated in cage studies with artificial diets were also shown to occur under field conditions.
The effects of tunicamycin and 20‐hydroxyecdysone on the cell surface properties of two insect cell lines
Authors: Jang Eric B. , Marks Edwin P.
Published: February 2005

The radiolabeled lectins, concanavalin A* and wheat germ agglutinin, were used to study surface properties of two insect cell lines. We also looked at the effects of tunicamycin and 20‐hydroxyecdysone on the binding of these lectins to one of the cell lines. Both UMBGE‐2 and CH‐MRRL cells bound both lectins, specifically. The CH‐MRRL cells showed an overall higher binding for the lectins than the UMBGE‐2 cells. This difference may account for some of the striking morphological difference seen between these cells. Tunicamycin and 20‐hydroxyecdysone decreased the binding of both [125I]‐Con A and [125I]‐WGA to CH‐MRRL cells. These results suggest that cell surface glycoproteins play a role in the modification of cellular morphology and in other hormone‐mediated physiological functions.
The action of iontophoretically applied L‐glutamate on an insect visceral muscle
Authors: Dunbar Stuart J. , Piek Tom
Published: February 2005

1) lontophoretic application of L‐glutamate was employed to study the distribution of glutamate receptors in the superior longitudinal (SL) muscles of the locust (Locusta migratoria) hindgut, in which spontaneous activity was inhibited using normal saline containing 5 mM MgCl2. 2) Junctional glutamate potentials with a rise time of 50–100 ms (peak) and a decay time of 250–400 ms were recorded at localized sites using ejection pulses in the range 5–10 nC. Most active sites were found in interfiber clefts and were spaced at about 250–300 μm intervals. 3) Desensitization of glutamate receptors occurred using ejection frequencies > 0.2 Hz. Desensitization could be irreversibly blocked using the lectin concanavalin A. 4) Depolarizing (D‐) and biphasic depolarizing/hyperpofarizing (DH ‐) extrajunctional glutamate potentials were observed using ejection pulses > 15 nC. 5) δ‐Philanthotoxin (δ‐PTX) at concentrations > 0.3 Uml−1 inhibited junctional glutamate potentials in a dose‐dependent manner, 50% inhibition was achieved using 0.45 Uml−1 δ‐PTX. 6) Subthreshold concentrations of proctolin (up to 5 × 10−10M) had no visible effect on glutamate potentials, suggesting that proctolin possibly does not act by modulating glutamate activity. 7) It is proposed that glutamate plays a transmitter role in SL muscles, while the role of proctolin is still unclear.
Treatment of larvae with repeated doses of precocene II induces precocious metamorphic changes in Spodoptera mauritia (Lepidoptera: Noctuidae)
Authors: Mathai Sam , Nair V. S. K.
Published: February 2005

In Spodoptera mauritia repeated daily treatments of larvae with 40 μg of precocene II throughout the fifth and sixth instar larval period had no effect on the larval‐larval period but prolonged larval‐pupal ecdysis. The resulting pupae showed precocious adult differentiation of mouth parts, wings, eyes, legs, and fat body.
Some pharmacological properties of neuromuscular transmission in the oviduct of the locust, Locusta migratoria
Authors: Lange Angela B. , Orchard Ian
Published: February 2005

The effects of various pharmacological agents on neurally evoked contractions of the visceral muscles of the oviduct of Locusta migratoria have been examined. The pentapeptide, proctolin, at low concentrations (10−11 M−10−10 M), induced an increase in the amplitude of neurally evoked contractions and basal tonus, and induced the appearance and increased the frequency of myogenic contractions. Glutamate, at 10−4 M, produced a small transient contraction which in some preparations was accompanied by a reduction in amplitude of neurally evoked contractions. Octopamine, at 10−6 M, reduced the amplitude of neurally evoked contractions and also resulted in a relaxation of the muscles. The octopaminergic effects were inhibited by the α‐aminergic antagonist phentolamine. Neurally evoked contractions were unaffected by dopamine, 5‐HT or the acetylcholine receptor antagonists atropine and hexamethonium. Acetylcholine increased the amplitude of neurally evoked contractions, but only at the high concentration of 10−3 M.

The possible role of proctolin and glutamate as excitatory neuro‐transmitters and the inhibitory action of octopamine is discussed.
Inhibitory probes of mitochondrial ATPase and the action of DDT
Authors: Dary C. C. , Cutkomp L. K.
Published: February 2005

Chemical inhibitors were used as probes of mitochondrial ATPase to determine the site of action of DDT on oligomycin‐sensitive mitochondrial ATPase (OS‐ATPase) using whole mitochondria isolated from red coxal muscle of the American cockroach. Several plotting procedures were employed to delineate the form of inhibition. Relative potency and joint action were used to detect similar action, synergism, and antagonism between DDT and the inhibitory probes DCCD, Nbf‐CI, and oligomycin. DDT demonstrated not (strictly) competitive kinetics and may be acting as an uncompetitive inhibitor. DDT and DCCD produced similar additive action. At limiting concentrations of DCCD, inhibition was reduced in the presence of DDT. Effects shown by oligomycin were not altered by DDT. DDT enhanced the effects of Nbf‐CI. These interactions, together with the demonstration of not (strictly) competitive kinetics, indicate that DDT may be acting on the membrane sector as an allosteric modifier.
Comparison of structurally analogous allatotoxins on the molting and morphogenesis of Rhodnius prolixus, and the reversal of ecdysial stasis by ecdysone
Authors: Garcia Eloi S. , de Azambuja Patricia , Bowers William S.
Published: February 2005

The allatotoxic effect of 3‐ethoxy‐4‐methoxy‐6‐iso‐pentenylphenol on nymphal molting and metamorphosis of Rhodnius prolixus was examined. Continuous contact treatment with IPP induced the formation of precocious adults and retarded molting or initiated a permanent ecdysial stasis. Insects treated with 7‐ethoxy‐6‐methoxy‐2,2‐dimethylchrornene were similarly affected. Ecdysone given orally counteracted the ecdysial stasis and also reduced the duration of the molting delay caused by IPP.
Comparative studies on pharmacokinetics and metabolism of the anti‐juvenile hormone precocene II
Authors: Haunerland Norbert H. , Bowers William S.
Published: February 2005

The cuticular penetration and pharmacokinetics of the anti‐juvenile hormone precocene II were determined in a sensitive species (Oncopeltus fasciatus) and an insensitive species (Heliothis zea). Precocene was sequestered by the fat body and slowly metabolized in Oncopeltus, but rapidly metabolized and excreted in Heliothis. Studies in vitro using inhibitors for cytochrome P‐450 and for cyt P‐450‐NADPH‐reductase, confirmed the anticipated detoxification of precocene by a mixed‐function oxidase via the 3,4‐epoxide. Use of the inhibitors in vivo had no influence on the metabolism of precocene.
Effect of metamorphosis on the major hemolymph proteins of the silkworm
Authors: Seong Su Ii , Park Kwang E. , Nagata Masao , Yoshitake Narumi
Published: February 2005

During the metamorphosis of the silkworm, Bombyx mori, three major hemolymph proteins (MHPs) (molecular weights 17,000, 25,000, 27,000) were detected and found to be distributed in the hemolymph and in the tissues of several organs, such as the fat body, midgut, ovary, testis, and even eggs. The MHPs in eggs gradually decreased and disappeared during embryogenesis. The formation, distribution, and utilization of MHPs in tissues other than the gonad, however, were not affected by sex.

Radioisotope experiments in vivo revealed that the MHPs were synthesized at an early period of the fifth larval instar. The synthesis of at least two of them occurred in the fat body. MHPs in the hemolymph entered the tissues at the onset of the larval‐pupal transformation. On the basis of their appearance, distribution, and depletion, the MHPs may be classified as reserve proteins which are synthesized in the larval stage and utilized later in the developmental stages.
The disappearance of injected prostaglandins from the circulation of adult female australian field crickets, Teleogryllus commodus
Authors: Stanley‐Samuelson David W. , Loher Werner
Published: February 2005

During the first 2 h following injection of 0.02 μCi of [14C]‐prostaglandin E2 into the abdomen of adult virgin female crickets, T. commodus, the concentration of radioactivity in the circulating hemolymph decreases. The reduction is associated with the increase in radioactivity in the Malpighian tubule/hindgut complex, ovaries, fat body, and, to a much smaller extent, ventral nerve cord and flight muscles. The finding that most, but not all, of the radioactivity in the hindgut is located in the contents of the lumen suggests that a high proportion of prostaglandins circulating in the hemolymph of T. commodus is eliminated by the usual excretory pathway. We suggest that the differential uptake of label from the circulating hemolymph by various tissues may be related to possible physiological functions that remain to be discovered.
Biosynthesis and distribution of ecdysone and 20‐OH‐ecdysone in Aedes aegypti
Authors: Borovsky Dov , Whisenton Lavern R. , Thomas Billy R. , Fuchs Morton S.
Published: February 2005

The distribution and biosynthesis of ecdysone and 20‐hydroxyecdysone (20‐OH‐ecdysone) was followed in sugar‐ and blood‐fed female Aedes aegypti. In both sugar‐ and early blood‐fed animals most of the ecdysteroid determined by radioimmunoassay was found outside the ovary. Twenty‐four to 40 h after blood feeding, however, ecdysteroid was distributed between ovary and carcass in the ratio of 1:1.5. Ecdysteroid titer reached a plateau between 18 to 40 h after the blood meal and decreased thereafter. Analysis of the ecdysteroid titer using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) revealed that both 20‐OH‐ecdysone and ecdysone were synthesized after the blood meal. The ratio of 20‐OH‐ecdysone to ecdysone remained essentially constant and fluctuated in parallel throughout egg development. Chromatography of the early ecdysteroid peak (8 h after feeding) using TLC and HPLC indicated that although it cross‐reacted with ecdysteroid antibodies, it did not have the same elution times as ecdysone and 20‐OH‐ecdysone and is, therefore, probably a precursor of these ecdysteroids. Injections of egg development neurosecretory hormone (EDNH) preparation purified to near homogeneity, into ligated abdomens, induced ecdysteroid synthesis only if the abdomens were first treated with methoprene (12.5 pg). Methoprene at this concentration did not stimulate ecdysteroid synthesis in these abdomens. When blood‐fed females were treated with [4‐14C] cholesterol and analyzed using TLC and HPLC procedures, both [14C]labeled ecdysone and [14C]labeled 20‐OH‐ecdysone were synthesized in the ratio of 1:1.5. This report is the first to show that both ecdysone and 20‐OH‐ecdysone are synthesized in vivo in female A. aegypti.
Linoleic acid biosynthesis in the pea aphid, Acyrthosiphon pisum (Harris)
Authors: De Renobales Mertxe , Ryan Robert O. , Heisler Charles R. , McLean Donald L. , Blomquist Gary J.
Published: February 2005

The pea aphid Acyrthosiphon pisum (Harris) incorporated [1‐14C]acetate into a phospholipid dienoic fatty acid in a time‐dependent manner. In 2‐h incubations, the incorporation of radioactivity into the 18:2 fraction was minimal, whereas after 45 h 18:2 was the major fatty acid labeled. Ozonolysis of the isolated dienoic fatty acid methyl ester followed by radio‐gas‐liquid chromatography showed that radioactivity was associated with fragments containing carbons 1–9 and 13–18. These data established the location of the double bonds in the 9,12 positions and indicated that the entire molecule was labeled from [1‐14C]acetate. Tetracycline‐treated aphids synthesized linoleic acid in the same proportions as untreated controls. Scanning electron microscopy showed that over 50% of the treated insects had greatly reduced numbers of intracellular symbiotes or lacked them or most of the existing symbiotes had an abnormal appearance. Therefore, we conclude that intracellular symbiotes are not involved in the biosynthesis of linoleic acid in the pea aphid.
Absence of female‐specific protein in the hemolymph of stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae)
Authors: Houseman Jon G. , Morrison P. E.
Published: February 2005

Changes, during the reproductive cycle, in fat body, hemolymph, and ovarian proteins of the stable fly Stomoxys calcitrans were characterized quantitatively and qualitatively using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Protein content of all three tissues increased after blood feeding. Fat body protein increased first, followed by hemolymph and ovarian proteins. SDS‐PAGE failed to identify vitellogenin in both female hemolymph and fat body samples. No single protein or group of proteins predominated at any stage of the reproductive cycle. Comparisons between male and female stable fly hemolymph and fat body proteins failed to detect female‐specific proteins. Female‐specific proteins, however, were detected in the hemolymph of four other species of Diptera.
The effect of L‐canavanine and L‐canaline on the hemolymph amino acid composition of the tobacco hornworm, Manduca sexta (L.) (Sphingidae)
Authors: Lenz Cynthia , Dahlman D. L. , Rosenthal G. A.
Published: February 2005

Tobacco hornworm larvae, Manduca sexta (L.) (Sphingidae), were administered L‐canaline either by parenteral injection or by dietary consumption. The overt toxicity and the alteration of hemolymph amino acids caused by these nonprotein amino acids were evaluated. The LD50 value for parenterally administered canavanine and canaline is 1.0 and 2.5 mg/g fresh body weight, respectively. A dietary concentration of 5.2 mM for canavanine and over 20 mM for canaline represent the respective LC50 values. A large percentage of the larvae reared on diets supplemented with additional arginine, ornithine, or 2,4‐diaminobutyric acid in addition to canavanine or canaline were unable to complete larval‐pupal ecdysis. These toxic effects were associated with a decreased glutamic acid hemolymph titer and dramatically elevated ornithine. On the other hand, larvae administered canavanine or canaline alone, either by dietary consumption or parenteral injection, experienced less drastic developmental aberrations. These symptoms were in some cases correlated with increased ornithine and glutamic acid titers. Evidence is presented that even a canavanine‐ and canaline‐sensitive insect such as M. sexta has a marked ability to eliminate these protective allelochemicals.
Effects of the tumor promoter 12‐0‐tetradecanoyl‐phorbol‐13‐acetate on the wasp, Bracon hebetor
Authors: Best R. G. , Grosch D. S.
Published: February 2005

A single oral dose of the tumor promoter, 12‐0‐tetradecanoyl‐phorbol‐13‐acetate (TPA), caused a rapid necrosis of the ovarioles, aberrations in the developmental sequence of oocytes, and a concomitant dose‐dependent decline in egg production in the wasp, Bracon hebetor. TPA and its metabolities were found to have a biological half life of 26.7 h, with a peak concentration in the ovarioles in 3 h. Damage to ovariole tissue was persistent despite the relatively short half life. Other tissues in the wasp were largely unaffected, although TPA induced lethargy that persisted until death. There was no shortening of life span. Inhibition of intercellular transport and metabolic cooperation may account for decreased fecundity and fertility, but interaction with a phorbol ester receptor is more likely to account for developmental changes and central nervous system poisoning.
Pupariation in flies: A tool for monitoring effects of drugs, venoms, and other neurotoxic compounds
Authors: Zdarek Jan , Fraenkel Gottfried
Published: February 2005

A complex Sarcophaga bullata pupariation assay was used to evaluate the neurotropic effects of several drugs, venoms, and insecticides. The assay consists of tests for (1) immediate effects on the intact larva, (2) effects on ligated (ie, isolated from the central nervous system) larval abdomens, (3) morphogenetic effects on the puparium, and (4) effects on stereotyped pupariation behavior. The latter are monitored barographically by recording changes in hemocoelic pressure. Of 62 compounds screened, 18 showed morphogenetic activity at a threshold dose of 5 μg or less, 11 at a dose of 50 μg, four at a dose of 100 μg, and 29 showed no morphogenetic activity. From a comparison of the putative pharmacological actions of the tested compounds with their morphogenetic effects, certain generalizations can be made: Agents that paralyze neuromuscular systems at the peripheral level (eg, tetrodotoxin), or suppress or modify basic motor patterns centrally (eg, veratrine sulphate), cause retention of larval morphological characters in the puparium. Compounds that stimulate convulsive contractions of segmental musculature (mostly cholinergic drugs like eserine sulphate, nicotine, organophosphate insecticides) cause retention of larval segmentation on longitudinally contracted puparia. Five compounds (venom of the scorpion, Leirus quinquestriatus, pyrethrins, protoveratrine A, and kainic and quisqualic acids) stimulate musculature of the denervated abdomen. Barographic monitoring of changes in pupariation behavior appears to be a most sensitive and informative test. It reveals great differences in the ways in which compounds producing seemingly identical morphogenetic effects affect and modify behavior, thus making pharmacological classification more accurate.
Effect of heat shock on protein synthesis in Locusta migratoria epidermis
Authors: Baldaia L. , Maisonhaute C. , Porcheron P. , Best‐Belpomme M.
Published: February 2005

Heat shock induced by an increase in temperature from 30°C to 47°C led to changes in protein synthesis in wing pads of the fifth larval instar of Locusta migratoria. Synthesis of heat shock proteins in the molecular weight range of 85,000, 70,000 and 18,000–22,000 was first detected at a threshold temperature of 45°C and was found to be highest at 47°C. A marked decline in the synthesis of many other proteins was also evident at 47°C. Recovery of general protein synthesis was observed when wing pads were shifted back to 30°C after a 2‐h heat shock at 47°C. Heat shock protein patterns in Locusta and Drosophila were compared.
N‐terminal amino acid sequence analysis of dihydrofolate reductase from methotrexate‐resistant mosquito cells
Authors: Johnston Anne M. , Fallon Ann Marie
Published: February 2005

Methotrexate‐resistant mosquito (Aedes albopictus) cell mutants were used as starting material for the purification of dihydrofolate reductase. Initial fractionation of a crude cell lysate by ammonium sulfate precipitation showed an enrichment of enzyme activity in the 60‐100% pellet. Purification of this ammonium sulfate fraction on a Sephadex G‐75 column prior to affinity chromatography was required for maximal yield of purified protein. A protein with dihydrofolate reductase activity and an Mr of 24,000 was eluted from a methotrexate‐agarose column with 0.5 M Tris‐HCl, pH 8.5, containing 2 mM folic acid. A 21‐fold increase in specific activity relative to that in the crude cell lysate was obtained, and amino acid sequence analysis confirmed that dihydrofolate reductase from the affinity column migrated as an electrophoretically pure band on SDS polyacrylamide gels. The initial 20 amino acid residues of the mosquito dihydrofolate reductase were identified by microsequencing and compared to those of murine, human, and bacterial dihydrofolate reductase molecules.
Pharmacology of octopamine‐, dopamine‐, and 5‐hydroxytryptamine‐stimulated cyclic amp accumulation in the corpus cardiacum of the american cockroach, Periplaneta americana L
Authors: Gole John W. D. , Orr Gregory L. , Downer Roger G. H.
Published: February 2005

A variety of potential receptor agonists were incubated with isolated corpora cardiaca from the American cockroach Periplaneta americana to determine their effects on cyclic AMP production in this gland. Octopamine, dopamine, and 5‐hydroxytryptamine elevated cyclic AMP levels in a dose‐dependent manner with estimated Ka values of 15.8, 1.7, and 1.1 m̈M, respectively, and their stimulation was found to be additive. Several vertebrate receptor antagonists were tested against the three amines and a preliminary pharmacological profile developed.
Hormonal control of ovarian development in the silkworm, Bombyx mori
Authors: Tsuchida Kozo , Nagata Masao , Suzuki Akinori
Published: February 2005

Hormonal control of ovarian development was examined in Bombyx mori. The weight of the ovary increased suddenly by 3 days after pupal ecdysis, and vitellogenin could be immunologically detected in the ovary at that time. The ecdysteroid titers during pupal‐adult development, quantified by radioimmunoassay, increased from day 0 to day 2. Ovarian development was arrested for a long period in brainless pupae and isolated pupal abdomens. Injection of 20‐hydroxyecdysone into such preparations stimulated development of the ovaries, and vitellogenin could be detected in ovaries 2 days after injection. The results suggest that 20‐hydroxyecdysone acts by stimulating the growth of ovary.
Tyrosinase catalyzed protein polymerization as an in vitro model for quinone tanning of insect cuticle
Authors: Sugumaran Manickam , Hennigan Brian , O'Brien Jean
Published: February 2005

The validity of Pryor's widely accepted quinone tanning hypothesis for the sclerotization of insect cuticle was examined using an in vitro model system. Quinones generated in situ by the oxidation of catechols with mushroom tyrosinase and molecular oxygen readily reacted with test proteins such as lysozyme, ribonuclease and cytochrome‐c, producing dimers, trimers, and higher oligomers. With the exception of 3,4‐dihydroxyphenylalanine, dopamine, and norepinephrine, most other catechols tested participated in protein polymerization. The inability of these three compounds to support oligomerization of test protein was attributed to their high rate of intramolecular cyclization reaction. Radioactive incorporation studies reveal the formation of catechol‐protomer adducts, as well as aryl‐protein crosslinks in the reaction mixture. The above results strongly support the quinone tanning hypothesis. Based on these findings, the mechanism of cuticular sclerotization is discussed.
Lipophorin in the larval and adult stages of Musca domestica
Authors: De Bianchi Antonio G. , Capurro Margareth De L. , Marinotti Osvaldo
Published: February 2005

The major Musca domestica hemolymph lipoprotein, lipophorin, was purified from larval and from adult animals. The housefly lipophorin is composed of two apoproteins, apolipophorin I (Mr ∽ 253,000) and apolipophorin II (Mr ∽ 85,000). The lipophorin contains about 3.9% carbohydrates and reacts positively with concanavalin A. The density of larval lipophorin is equal to 1.152 g/ml and of adult lipophorin to 1.106 g/ml. The amount of lipophorin per animal increases during the larval stage, is constant during pupal stage, and suffers a great reduction at the pharate adult stage. The amount of lipophorin remains stable during the whole first gonotrophic cycle of the housefly. Lipophorin is not detected in the eggs of this insect.
Purification and properties of a factor from insect hemolymph that promotes multicellular vesicle formation in vitro
Authors: Ferkovich S. M. , Oberlander H. , Dillard C. , Leach C. E.
Published: February 2005

A vesicle promoting factor (VPF) in an insect cell line IAL‐TND1 was partially purified from larval hemolymph of the cabbage looper, Trichoplusia ni Hübner. The polypeptide had a molecular weight estimated to be between 20.5 kD and 37.5 kD by gel permeation, 22.5 kD by the Ferguson plot on nondenaturing polyacrylamide gel electrophoresis, and 16.88 kD on sodium dodecyl sulfate PAGE. VPF fractions were isolated from hemolymph by gel permeation on Fractogel® and were either subjected to chromatofocusing or preparative isoelectric focusing. After the gel permeation step, the VPF polypeptide was highly unstable during the separation and storage. The two active fractions from the isoelectric separations had isoelectric points of 6.21 and 6.36 and had specific activities of 34 and 32 vesicles/μg protein per test culture chamber. The percentage of total larval equivalent hemolymph proteins found in these fractions was less than 1%. Chromatofocusing technique also yielded an active fraction containing a single band on nondissociating electrophoresis that had VPF activity. This band had an isoelectric point of 6.60 but had a lower specific activity of three vesicles/μg protein in the VPF cell bioassay.
Evidence for a sex pheromone metabolizing cytochrome P‐450 mono‐oxygenase in the housefly
Authors: Ahmad Sami , Kirkland Kenneth E. , Blomquist Gary J.
Published: February 2005

Direct evidence is presented for the role of a cytochrome P‐450 monooxygenase (called mixed‐function oxidase, or polysubstrate mono‐oxygenase, PSMO) in the metabolism of the sex pheromone (Z)‐9‐tricosene to its corresponding epoxide and ketone in the housefly. A secondary alcohol, most likely an intermediate in the conversion of the alkene to the ketone, was also tentatively identified. The results of in vivo and in vitro experiments showed that the PSMO inhibitors, piperonyl butoxide (PB) and carbon monoxide, markedly inhibited the formation of epoxide and ketone from (9,10‐3H) (Z)‐9‐tricosene. An examination of the relative rates of (Z)‐9‐tricosene metabolism showed that males exhibited a higher rate of metabolism than females with the antennae of males showing the highest activity of any tissue/organ examined. The major product from all tissues/organs was the epoxide. Data from experiments with subcellular fractions showed that the microsomal fraction had the majority of enzyme activity, which was strongly inhibited by PB and CO and required NADPH and O2for activity. A carbon monoxide difference spectrum with reduced cytochrome showed maximal absorbance at 450 nm and allowed quantification of the cytochrome P‐450 in the microsomal fraction of 0.410‐nmol cytochrome P‐450 mg−1protein. Interaction of (Z)‐9‐tricosene with the cytochrome P‐450 resulted in a type I spectrum, indicating that the pheromone binds to a hydrophobic site adjacent to the heme moiety of the oxidized cytochrome P‐450.
Pharmacology of [3H]mianserin binding in the nerve cord of the American cockroach, Periplaneta americana
Authors: Minhas Nailah , Gole John W. D. , Orr Gregory L. , Downer Roger G. H.
Published: February 2005

The binding of [3H]mianserin to homogenates of cockroach nerve cord was investigated, using a centrifugal binding assay. Nonspecific binding was defined as binding in the presence of 0.1 μM phentolamine. Specific binding was saturable and indicated a high affinity site (KD = 39.6 nM and Bmax = 0.8 fmol μGg−1) and a low affinity site (KD = 648.9 nM and Bmax = 5.9 fmol μg−1). The binding was reduced or abolished by heat, detergents, trypsin and HgCl2. Incorporation of CaCl2 or NaCl into the medium reduced binding in a dose‐dependent fashion, whereas MgCl2 and NaF inhibited binding in a biphasic manner. The stable GTP analog, 5′‐guanylylimidodiphosphate, reduced the high affinity binding at 10 μM. Phentolamine, D,L‐octopamine, demethylchlordimeform (DCDM), clonidine, and D,L‐synephrine are effective displacers of [3H]mianserin‐binding, whereas dopamine, acetylcholine, serotonin (5‐HT), and histamine show poor or no displacement. The pharmacological properties of octopamine‐sensitive adenylate cyclase stimulation in cockroach nerve cord are similar to those of [3H]mianserin‐binding.
Cell culture techniques for studying insect cuticle
Authors: Marks Edwin P. , Ward Gordon B.
Published: February 2005

Evidence that biosynthetic pathways critical to the formation of insect cuticle are retained in continuous insect cell lines opens new possibilities for research on the cuticle system. Recent findings indicate that chitin, molting hormone, and catecholamines are all produced by a vesicle cell line derived from embryos of the cockroach Blattella germanica. The chitin that is formed by this cell line is particulate and does not show the characteristic featherlike crystalline structure found in mature cuticle. The molting hormone is produced as ecdysone and is released into the culture medium. The addition of 20‐hydroxyecdysone to the cultures increases the production of chitin fourfold. These responses are similar to those found in insect organ cultures.
Isolation and fluorescence studies on a lipophorin from the weevil diaprepes abbreviatus
Authors: Shapiro Jeffrey P.
Published: February 2005

Lipophorin was isolated from larvae of a root weevil, Diaprepes abbreviatus (Coleoptera: Curculionidae), using density gradient ultracentrifugation. D. abbreviatus lipophorin contained two apoproteins, apolipophorin‐I (Mr = 226,000) and apolipophorin‐II (Mr = 72,100) and had a density of 1.08. Relative to other larval lipophorins, D. abbreviatus lipophorin contained little cysteine (determined as cysteic acid) and methionine. Fluorescence spectroscopy of intrinsic tyrosine and tryptophan residues excited at 290 nm revealed a single broad emission peak at 330 nm. Upon denaturing and delipidating lipophorin in guanidine HCl, this peak resolved into two peaks with maxima at 305 and 350 nm. Excitation spectra suggested that the two peaks were due to tyrosine and tryptophan, respectively. Fluorescence quenching agents, iodide and acrylamide, were used to determine accessibility of tyrosine and tryptophan residues to the aqueous environment. Iodide, a polar quenching agent, did not quench fluorescent emission from native lipophorin; quenching by iodide increased to moderate levels when lipophorin was denatured in guanidine HCl. Acrylamide quenched the fluorescence of native lipophorin moderately and very efficiently quenched fluorescence of denatured lipophorin. No difference was observed between fluorescence quenching of denatured vs. denatured and delipidated lipophorin by either iodide or acrylamide.
Subcellular distribution and activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase in the southern armyworm, Spodoptera eridania
Authors: Ahmad Sami , Pritsos Chris A. , Bowen Susan M. , Heisler Charles R. , Blomquist Gary J. , Pardini Ronald S.
Published: February 2005

In mid‐fifth‐instar larvae of the southern armyworm, Spodoptera eridania, the subcellular distribution of four antioxidant enzymes—superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR)—were examined. Two‐thirds (4.26 units ·mg protein−1) of the SOD activity was found in the cytosol, and one‐thirds (2.13 units ·mg protein−1) in the mitochondria. CAT activity was unusually high and not restricted to the microsomal fraction where peroxisomes are usually isolated. The activity was distributed as follows: cytosol (163 units) mitochondria (125 units) and microsomes (119 units). Similar to CAT, the subcellular compartmentalization of both GPOX and GR was unusual. No activity was detected in the cytosol, but in mitochondria and microsomes, GR levels were 5.49 and 3.09 units. Although GPOX activity exhibited 14–16‐fold enrichment in mitochondria and microsomes, respectively, over the 850g crude homogenate, the level was negligible (mitochondria = 1.4 × 10−3 units; microsomes = 1.6 × 10−3 units), indicating that this enzyme is absent. The unusual distribution of CAT has apparently evolved as an evolutionary answer to the absence of GR from the cytosol, and the lack of GPOX activity.