Comparison of two sterile Mediterranean fruit fly (Diptera: Tephritidae) strains released in California's Preventative Release Program

At Mediterranean fruit fly rearing centers, genetic sexing strains such as the male-only temperature sensitive lethal (tsl) strains, are replacing many of the older bisexual strains, such as Maui-93. Three tests were performed to compare the performance of the Vienna-4 (which carries tsl) and Maui-93 strains. Using standard laboratory quality control tests, the Maui-93 strain had higher percent fliers and a lower percentage deformed and partially eclosed flies. Analyses of data from field mark-recapture tests with Jackson traps failed to find significant differences in recapture numbers between the two strains. In field cage survivorship tests, the Vienna-4 strain showed significantly lower mortality in comparison to the Maui-93 strain. These results suggest that although the quality of Vienna-4 flies is lower than Maui-93 at the time of adult emergence, it is comparable with or better than Maui-93 several days later, because weaker flies have been eliminated from the cohort. The benefits of releasing only sterile males, instead of both males and females, was not factored into the quality assessment in this study, but would be an additional benefit in using a strain that carries a tsl mutation.     

Flower choice by naïve young crab spiders and the effect of subsequent experience

Initial responses of na?ve individuals to critical environmental stimuli provide important information about the innate contribution to behaviour, and subsequent responses to the same stimuli may show the role of experience in mediating those initial responses. To test the role of these factors, I measured initial patch choices and giving-up responses of just-emerged, na?ve, second-instar crab spiders, Misumena vatia, on several hunting sites they encountered after leaving their natal nests. In follow-up tests I measured the effects of these experiences on subsequent patch choice decisions. The choice of hunting sites is a vital decision at all stages of the life cycle for sit-and-wait predators such as Misumena. In their initial tests these spiderlings remained more frequently on goldenrod (Solidago spp.) flowers than on green or yellow goldenrod buds, a preference they retained through tests run on 5 consecutive days. Individuals on green and yellow buds shifted sites more quickly and frequently than those from flowers, and made most of these moves to flowers, which attracted many more prey than did buds. These differences were not affected by age, energetic condition, or loss of information over the period of the experiment. Once spiderlings moved from buds, they showed a high, increasing tendency to move from buds in subsequent runs, those from flowers showed a consistently low tendency. These results suggest that spiderlings retain their innate behavioural patterns through the second instar, but that experience also plays a modest role in patch choice at this stage. Copyright 2000 The Association for the Study of Animal Behaviour.     

Effect of Fas+ and Fas− Target Cells on the Ability of NK Cells to Repeatedly Fragment DNA and Trigger Lysis via the Fas Lytic Pathway

We and others have recently shown that human NK cells express the Fas ligand (FasL) constitutively and that they can trigger the lysis of Fas positive (Fas+) target cells (TC) by apoptosis. We have also previously demonstrated that NK cells exposed to sensitive TC temporarily lose their ability to lyse sensitive TC via the granule-mediated pathway and that this loss is recovered when inactivated NK cells (NKi) are incubated in medium supplemented with IL-2, IL-12 or IL-15. In this study, we investigated the fate of the Fas-lytic pathway in NK cells exposed to either Fas+ or Fas– TC. To this end, we exposed NK cells to Jurkat (Fas–) or Jurkat (Fas+) TC for up to 6 h, separated NK cells from the TC and assessed the residual lytic activity against K562, a traditional human NK cell target, Jurkat Fas+ and Jurkat Fas– TC. Fas lytic activity was determined in calcium free medium, in the presence or absence of two distinct Fas-blocking monoclonal antibodies and a Fas.Fc fusion protein. In parallel experiments, the extent of DNA fragmentation in the three TCs was also assayed by the JAM test. Our results indicate that: (i) NK cells exposed to susceptible Fas+ TC temporarily lose most of their lytic potential due to the granule-mediated pathway, while only partially losing the Fas-lytic pathway. They also partially lose their ability to fragment DNA. (ii) NK cells exposed to Fas+ TC completely recover the Fas lytic pathway and the ability to fragment DNA via the Fas/Fas ligand when incubated in medium supplemented with IL-2 for 18 h.     

High-performance liquid chromatographic method for measurement of cytochrome P450-mediated metabolism of 7-ethoxy-4-trifluoromethylcoumarin

An HPLC method for analysis of deethylation of 7-ethoxy-4-trifluoromethylcoumarin (ETFMC), a substrate of various enzymes of the cytochrome P450 superfamily, was developed. ETFMC was incubated at 37°C with human hepatic microsomes or microsomes prepared from a lymphoblastoid cell line that expresses human CYP2B6. Under these conditions, the highly fluorescent metabolite 7-hydroxy-4-trifluoromethylcoumarin (HTFMC) is formed. The metabolite was analyzed by reversed-phase HPLC with fluorescence detection. The limits of detection of the metabolite were 5.0 fmol per injection, a sensitivity at least one order of magnitude greater than the standard method, which does not involve HPLC. This method will be of great utility when quantities of microsomal protein from cell lines expressing human CYP enzymes are limited.     

Posture Affects How Robots and Infants Map Words to Objects

For infants, the first problem in learning a word is to map the word to its referent; a second problem is to remember that mapping when the word and/or referent are again encountered. Recent infant studies suggest that spatial location plays a key role in how infants solve both problems. Here we provide a new theoretical model and new empirical evidence on how the body – and its momentary posture – may be central to these processes. The present study uses a name-object mapping task in which names are either encountered in the absence of their target (experiments 1–3, 6 & 7), or when their target is present but in a location previously associated with a foil (experiments 4, 5, 8 & 9). A humanoid robot model (experiments 1–5) is used to instantiate and test the hypothesis that body-centric spatial location, and thus the bodies’ momentary posture, is used to centrally bind the multimodal features of heard names and visual objects. The robot model is shown to replicate existing infant data and then to generate novel predictions, which are tested in new infant studies (experiments 6–9). Despite spatial location being task-irrelevant in this second set of experiments, infants use body-centric spatial contingency over temporal contingency to map the name to object. Both infants and the robot remember the name-object mapping even in new spatial locations. However, the robot model shows how this memory can emerge –not from separating bodily information from the word-object mapping as proposed in previous models of the role of space in word-object mapping – but through the body’s momentary disposition in space.     

Nonflowering plants possess a unique folate-dependent phenylalanine hydroxylase that is localized in chloroplasts

Tetrahydropterin-dependent aromatic amino acid hydroxylases (AAHs) are known from animals and microbes but not plants. A survey of genomes and ESTs revealed AAH-like sequences in gymnosperms, mosses, and algae. Analysis of full-length AAH cDNAs from Pinus taeda, Physcomitrella patens, and Chlamydomonas reinhardtii indicated that the encoded proteins form a distinct clade within the AAH family. These proteins were shown to have Phe hydroxylase activity by functional complementation of an Escherichia coli Tyr auxotroph and by enzyme assays. The P. taeda and P. patens AAHs were specific for Phe, required iron, showed Michaelian kinetics, and were active as monomers. Uniquely, they preferred 10-formyltetrahydrofolate to any physiological tetrahydropterin as cofactor and, consistent with preferring a folate cofactor, retained activity in complementation tests with tetrahydropterin-depleted E. coli host strains. Targeting assays in Arabidopsis thaliana mesophyll protoplasts using green fluorescent protein fusions, and import assays with purified Pisum sativum chloroplasts, indicated chloroplastic localization. Targeting assays further indicated that pterin-4a-carbinolamine dehydratase, which regenerates the AAH cofactor, is also chloroplastic. Ablating the single AAH gene in P. patens caused accumulation of Phe and caffeic acid esters. These data show that nonflowering plants have functional plastidial AAHs, establish an unprecedented electron donor role for a folate, and uncover a novel link between folate and aromatic metabolism.     

SMN and the Gemin proteins form sub-complexes that localise to both stationary and dynamic neurite granules

Childhood spinal muscular atrophy (SMA) is caused by a reduction in survival motor neuron (SMN) protein. SMN is expressed in every cell type, but it is predominantly the lower motor neurones of the spinal cord that degenerate in SMA. SMN has been linked to the axonal transport of β-actin mRNA, a breakdown in which could trigger disease onset. It is known that SMN is present in transport ribonucleoproteins (RNPs) granules that also contain Gemin2 and Gemin3. To further characterise these granules we have performed live cell imaging of GFP-tagged SMN, GFP-Gemin2, GFP-Gemin3, GFP-Gemin6 and GFP-Gemin7. In all, we have made two important observations: (1) SMN granules appear metamorphic; and (2) the SMN-Gemin complex(es) appears to localise to two distinct subsets of bodies in neurites; stationary bodies and smaller dynamic bodies. This study provides an insight into the neuronal function of the SMN complex.