Rhesus monkey behavior during exposure to high-peak-power 5.62-GHz microwave pulses

Limits on the exposure to high-peak-power, short-duration microwave pulses have only recently been adopted. Additional data, however, are needed to understand the effects that may be produced by exposure to high-peak-power pulsed microwaves. Four male rhesus monkeys (Macaca mulatta) were trained on an operant task for food pellet reward to investigate the behavioral effects of very high-peak-power 5.62 GHz microwaves. The operant task required monkeys to pull one plastic lever on a variable interval schedule (VI-25 s) and then respond to color signals and pull a second lever to obtain food. The monkeys were conditioned to perform a color discrimination task using one of three colors displayed by a fiber-optic cable. A red signal was the discriminative stimulus for responding on the first lever. A response on the second lever when a green signal was presented (1 s duration) delivered a food pellet. If a response on the second lever was made in the presence of a white signal, a 30-s timeout occurred. While performing the behavioral task, the monkeys were exposed to microwave pulses produced by either a military radar (FPS-26A) operating at 5.62 GHz or the same radar coupled to a Stanford linear energy doubler (SLED) pulse-forming device (ITT-2972) that enhanced peak power by a factor of nine by adding a high power pulse to the radar pulse. The effects of both types of pulses were compared to sham exposure. Peak field power densities tested were 518, 1270, and 2520 W/cm² for SLED pulses and 56, 128, and 277 W/cm² for the radar pulses. The microwave pulses (radar or SLED) were delivered at 100 pps (2.8 μs radar pulse duration, ≈ 50 ns SLED pulse duration) for 20 min and produced averaged whole-body SARs of 2,4, or 6 W/kg. Compared to sham exposures, significant alterations of lever responding, reaction time, and earned food pellets occurred during microwave exposure at 4 and 6 W/kg but not 2 W/kg. There were no differences between radar or SLED pulses in producing behavioral effects. ©1994 Wiley-Liss, Inc.

1 This is a US Government work and, as such, is in the public domain in the United States of America.

     

Studies with Differentially Labeled [11C]Cocaine, [11C]Norcocaine, [11C]Benzoylecgonine,and [11C]-and 4′-[18F]Fluorococaine to Probe the Extent to Which [11C]Cocaine Metabolites Contribute to PET Images of the Baboon Brain

Abstract: The psychostimulant drug of abuse, cocaine (benzoylecgonine methyl ester), is rapidly metabolized by cleavage of its two ester groups, to give benzoylecgonine (BE) and ecgonine methyl ester, and by N-demethylation, to give N-norcocaine (NC). The recent use of [N-methyl-¹¹CH₃]cocaine to image brain cocaine binding sites with positron emission tomography (PET) raises the question of whether PET images partially reflect the distribution and kinetics of labeled cocaine metabolites. We prepared [O-metty/-¹¹CH₃]cocaine by methylation of the sodium salt of BE with [¹¹C]CH₃l, and showed that PET baboon brain scans, as well as regional brain kinetics and plasma time-activity curves corrected for the presence of labeled metabolites, are nearly identical to those seen with [N-methyl-¹¹CH₃]cocaine. This strongly suggests that ¹¹C metabolites do not significantly affect PET images, because the metabolite pattern is different for the two labeled forms of cocaine. In particular, nearly half the ¹¹C in blood plasma at 30 min was [¹¹C]CO₂ when [N-methy/-¹¹CH₃]cocaine was administered, whereas [¹¹C]CO₂ was not formed from [O-methy/-¹¹CH₃]cocaine. Only a trace of [¹¹C]NC was detected in plasma after [O-methyl-¹¹CH₃]cocaine administration. Nearly identical brain PET data were also obtained when 4′-[N-methy/-¹¹CH₃]fluorococaine and 4′-[¹⁸F]fluoro-cocaine (prepared by nucleophilic aromatic substitution from [¹⁸F]fluoride-and 4′-nitrococaine) were compared with [N-methy/-¹¹CH₃]cocaine. In vitro assays with rat brain membranes showed that cocaine and 4′-fluoroco-caine were equipotent at the dopamine reuptake site, but that 4′-fluorococaine was about 100 times more potent at the 5-hydroxytryptamine reuptake site. The studies with positron-emitting 4′-fluorococaines thus support the lack of significance of labeled metabolites or of binding to 5-hydroxytryptamine reuptake sites to PET images taken with [N-methy/-¹¹CH₃]cocaine. [¹¹C]NC prepared by O-methylation of norbenzoylecgonine gave PET images with preferential uptake in striatum, but slower clearance from all brain regions than [O-methy/-¹¹CH₃]cocaine. [¹¹C]BE prepared by N-methylation of norbenzoylecgonine did not show brain uptake.     

Frank Macfarlane Burnet and the immune self

This essay is excerpted from Alfred I. Tauber,The Immune Self: Theory or Metaphor? (New York and Cambridge: Cambridge University Press, 1994).     

The new class 11 transposon Tn163 is plasmid-borne in two unrelated Rhizobium leguminosarum biovar viciae strains

Tn163 is a transposable element identified in Rhizobium leguminosarum bv. viciae by its high insertion rate into positive selection vectors. The 4.6 kb element was found in only one further R. leguminosarum bv. viciae strain out of 70 strains investigated. Both unrelated R. leguminosarum bv. viciae strains contained one copy of the transposable element, which was localized in plasmids native to these strains. DNA sequence analysis revealed three large open reading frames (ORFs) and 38 bp terminal inverted repeats. ORF1 encodes a putative protein of 990 amino acids displaying strong homologies to transposases of class 11 transposons. ORF2, transcribed in the opposite direction, codes for a protein of 213 amino acids which is highly homologous to DNA invertases and resolvases of class II transposons. Homology of ORF1 and ORF2 and the genetic structure of the element indicate that Tn163 can be classified as a class II transposon. It is the first example of a native transposon in the genus Rhizobium. ORF3, which was found not to be involved in the transposition process, encodes a putative protein (256 amino acids) of unknown function. During transposition Tn163 produced direct repeats of 5 bp, which is typical for transposons of the Tn3 family. However, one out of the ten insertion sites sequenced showed a 6 by duplication of the target DNA; all duplicated sequences were A/T rich. Insertion of Tn163 into the sacB gene revealed two hot spots. Chromosomes of different R. leguminosarum bv. viciae strains were found to be highly refractory to the insertion of Tn163.     

Evaluation of the palatability of chrysomelid larvae containing anthraquinones to birds

Chrysomelid larvae of the subfamily Galerucinae, tribe Galerucini, are known to contain 1,8-dihydroxylated 9,10-anthraquinones. Since nonhydroxylated 9,10-anthraquinone is the active agent in several commercial products sold to protect seeds against birds, we suggested that the naturally occurring dihydroxylated anthraquinones of galerucine larvae may also act as protective devices against bird predation. Tits (Parus spp.) are potential predators of larvae of the tansy leaf beetle, Galeruca tanaceti, and the elm leaf beetle, Xanthogaleruca luteola. To investigate the palatability of these chrysomelid larvae to birds, we offered them with mealworms and Calliphora pupae, respectively, as controls in dual choice bioassays to eight singly kept, naive tits (five P. major and three P. ater individuals). The bioassays were limited to 5 days, during which larvae were offered daily for 2 h (X. luteola) and 3 h (G. tanaceti), respectively. Every day, the birds significantly avoided uptake of G. tanaceti and X. luteola. More than 98% of the control food was consumed daily, whereas the percentage of chrysomelid larvae totally eaten never surpassed 6.6% for G. tanaceti and 51.8% for X. luteola. In order to determine whether this avoidance was due to the anthraquinones of the chrysomelid larvae, mealworms and Calliphora pupae, respectively, were treated with these compounds in concentrations equivalent to the natural ones. Dual choice bioassays with treated and untreated prey were conducted, again for 5 days with a daily 2- or 3-h test period, respectively. The tits ate all or nearly all treated and untreated food items every day. However, during the 5-day test period the tits learnt to take up the control insects significantly earlier than the treated ones; the food containing anthraquinones was not consumed as readily as the control, which suggest aversive learning based on distastefulness. The efficiency of anthraquinones in protecting galerucine larvae against bird predation is discussed with special respect to learning behavior and factors which might delay or mask learning of avoidance.     

Bi-directional transfer of nitrogen between alfalfa and bromegrass: Short and long term evidence

Transfer of N from legumes to associated non-legumes has been demonstrated under a wide range of conditions. Because legumes are able to derive their N requirements from N₂ fixation, legumes can serve, through the transfer of N, as a source of N for accompanying non-legumes. Studies, therefore, are often limited to the transfer of N from the legume to the non-legume. However, legumes preferentially rely on available soil N as their source of N. To determine whether N can be transferred from a non-legume to a legume, two greenhouse experiments were conducted. In the short-term N-transfer experiment, a portion of the foliage of meadow bromegrass (Bromus riparius Rhem.) or alfalfa (Medicago sativa L.) was immersed in a highly labelled ¹⁵N-solution and following a 64 h incubation, the roots and leaves of the associated alfalfa and bromegrass were analyzed for ¹⁵N. In the long-term N transfer experiment, alfalfa and bromegrass were grown in an ¹⁵N-labelled nutrient solution and transplanted in pots with unlabelled bromegrass and alfalfa plants. Plants were harvested at 50 and 79 d after transplanting and analyzed for ¹⁵N content. Whether alfalfa or bromegrass were the donor plants in the short-term experiment, roots and leaves of all neighbouring alfalfa and bromegrass plants were enriched with ¹⁵N. Similarly, when alfalfa or bromegrass was labelled in the long-term experiment, the roots and shoots of neighbouring alfalfa and bromegrass plants became enriched with ¹⁵N. These two studies conclusively show that within a short period of time, N is transferred from both the N₂-fixing legume to the associated non-legume and also from the non-legume to the N₂-fixing legume. The occurrence of a bi-directional N transfer between N₂-fixing and non-N₂-fixing plants should be taken into consideration when the intensity of N cycling and the directional flow of N in pastures and natural ecosystems are investigated.     

Tissue and subcellular immunolocalisation of enzymes of lignin synthesis in differentiating and wounded hypocotyl tissue of French bean (Phaseolus vulgaris L.)

Antisera raised againstl-phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), and a cationic cell-wall peroxidase, which had all been purified from suspension-cultured cells of French bean, have been used to carry out immunogold localisations in the growing plant. Immunoglobulin-G fractions were prepared from each antiserum and used to study the distribution of the enzymes in differentiating and wounded hypocotyls by immunogold techniques and visualisation by both light and electron microscopy. Following silver enhancement to amplify the signal, proteins were detected by confocal microscopy in both developing (pre-xylem/ phloem) and later metaxylem stelar tissue.l-Phenylalanine ammonia-lyase and C4H also accumulated in cells adjacent to metaxylem, presumably involved in maintaining a supply of phenylpropanoid precursors to the enucleated xylem for further lignin synthesis. In these cells, PAL subunits were cytosolic although some were associated with endomembrane. Cinnamate-4-hydroxylase was wholly associated with membrane and particularly high concentrations were found in the Golgi bodies. The cationic peroxidase accumulated in xylem at sites of secondary thickening and in the middle lamella. The three proteins are also involved in defensive lignification. Thus when visualised by light microscopy, PAL and C4H were seen to accumulate to high levels throughout the cell types in wound sites and especially in the epidermal cells. An even more intense general distribution was found upon hyperinduction of wounded cells with-aminooxy--phenylpropionic acid. At the subcellular level, PAL was found to be localised in the cytosol in the wounded cells; however, because of the loss of membrane through mechanical damage, association with membrane structures, particularly endoplasmic reticulum, in unwounded cells is not entirely ruled out. Cinnamate-4-hydroxylase was associated with membranes when these were preserved. In wounded tissue, the peroxidase was found at the growing edges of tylose-like structures in the vascular xylem.Abbreviations AOPP -aminooxy--phenylpropionic acid - C4H cinnamic acid-4-hydroxylase - CHS chalcone synthase - GRP glycine-rich glycoprotein - HRGP hydroxyproline-rich glycoprotein - Ig immunoglobulin - PAL phenylalanine ammonia-lyase G.P.B. thanks the Agicultural and Food Research Council for support.     

Phylogenetic and biogeographic implications of chloroplast DNA variation in Picea

Purified chloroplast DNA ( cp DNA) extracts from 31 species of Picea and two species of Pinus (P. sylvestris and P. cembra ) were digested with eight restriction endo-nucleases, separated by electrophoresis and scored for restriction fragment length polymorphisms. The resulting data was analyzed phenetically and cladistically. The phenetic analysis indicated lower levels of cpDNA differentiation within Picea than within Pinus and lower levels of differentiation among Eurasian than among North-American Picea species. The cladistic analysis, using Pinus sylvestris as an outgroup, suggested monophyly for Picea and resolved several monophyletic groups among the 31 species of Picea . An assessment of biogeographic events, based on the cladogram, suggests that Picea originated in North-America and that the colonization of Eurasia occurred through separate, intercontinental migrations.