Patterning during pupal commitment of the epidermis in the butterfly, Precis coenia: the role of intercellular communication

The commitment of cells to pupal development in the larvae of holometabolous insects can be prevented by treatment with juvenile hormone (JH) or a JH mimic during a critical period early in the last larval instar. By treating larvae of different ages with a JH mimic, pupal commitment of the epidermis of the butterfly, Precis coenia, was found to occur in a strict temporal and spatial progression, which was serially homologous and occurred independently in each segment. The mechanism underlying this sequence of pupal commitment was examined by cauterizing regions of the epidermis to observe the effects of local ablation on the pattern of pupal commitment revealed by treatment with the JH mimic. Cautery of the segmental site of origin of pupal commitment, the dorsal midline, suppressed pupal commitment in the rest of the operated segment, indicating that the midline has a special effect on commitment of the rest of the segment. Cautery off the midline produced asymmetries in the pattern of pupal commitment; when placed close to the midline, such cauteries prevented pupal commitment in the region "downstream" of the cautery, suggesting that a signal (diffusible or transducible) emanates from the midline. Finally, cautery of a circle around the midline inhibited pupal commitment only outside the circle, showing that cautery could act as a barrier to the passage of a signal coming from the midline. These results suggest that inductive as well as hormonal signals are involved in the regulation of pupal commitment in the epidermis of the lepidopteran, P. coenia.     

Aggregation-dependent turnover of flagellar adhesion molecules in chlamydomonas gametes

Previous studies on flagellar adhesion in chlamydomonas (Snell, W. and S. Roseman. 1979. J. Biol. Chem. 254:10820-10829.) have shown that as gametes adhere to flagella isolated from gametes of the opposite mating type, the adhsiveness of the added flagella but not of the gametes is lost. The studies reported here show that the addition of protein synthesis inhibitors (cycloheximide [CH] or anisomycin) to the medium of such cell- flagella mixtures causes the cells to lose their adhesiveness. This loss, however, occurs only after the cells have interacted with 4-8 flagella/cell and does not occur if the cells are kept in CH (7 h) without aggregating. The availability of an impotent (imp) mating type plus (MT(+)) mutant (provided by U.W. Goodenough), which adheres but is unable to undergo the fusion that normally follows adhesion, made it possible to determine whether a similar loss of adhesiveness occurs in mixtures of matting type minus (mt(-)) and imp mt(+) gametes. In the absence of inhibitor, mt(-) and imp mt(+) gametes adhered to each other (without fusing) for several hours; however, in the presence of CH or anisomycin, the gametes began to de-adhere 35 min after mixing, and, by 90 min, 100 percent of the cells were single again. This effect was reversible, and the rapid turnover of cells were single again. This effect was reversible, and the rapid turnover of molecules involved in adhesion occurred only during adhesion inasmuch as gametes pretreated for 4 h with CH were able to aggregate in CH for the same length of time as nonpretreated cells aggregated in CH. By the addition of CH at various times after the mt(-) and imp mt(+) gametes were mixed, measurements were made of the “pool size” of the molecules involved in adhesion. The pool reached a minimum after 25 min of aggregation, rapidly increased for the next 25 min, and then leveled off at the premixing level. These results suggest that flagellar adhesion in chlamydomonas causes modification of surface molecules (receptors, ligands), which brings about their inactivation and stimulates their replacement.     

The effectiveness of flower strips and hedgerows on pest control,pollination services and crop yield: a quantitative synthesis

Floral plantings are promoted to foster ecological intensification of agriculture through provisioning of ecosystem services. However, a comprehensive assessment of the effectiveness of different floral plantings, their characteristics and consequences for crop yield is lacking. Here we quantified the impacts of flower strips and hedgerows on pest control (18 studies) and pollination services (17 studies) in adjacent crops in North America, Europe and New Zealand. Flower strips, but not hedgerows, enhanced pest control services in adjacent fields by 16% on average. However, effects on crop pollination and yield were more variable. Our synthesis identifies several important drivers of variability in effectiveness of plantings: pollination services declined exponentially with distance from plantings, and perennial and older flower strips with higher flowering plant diversity enhanced pollination more effectively. These findings provide promising pathways to optimise floral plantings to more effectively contribute to ecosystem service delivery and ecological intensification of agriculture in the future.     

Urban land use limits regional bumble bee gene flow

Potential declines in native pollinator communities and increased reliance on pollinator‐dependent crops have raised concerns about native pollinator conservation and dispersal across human‐altered landscapes. Bumble bees are one of the most effective native pollinators and are often the first to be extirpated in human‐altered habitats, yet little is known about how bumble bees move across fine spatial scales and what landscapes promote or limit their gene flow. In this study, we examine regional genetic differentiation and fine‐scale relatedness patterns of the yellow‐faced bumble bee, Bombus vosnesenskii, to investigate how current and historic habitat composition impact gene flow. We conducted our study across a landscape mosaic of natural, agricultural and urban/suburban habitats, and we show that B. vosnesenskii exhibits low but significant levels of differentiation across the study system (F_ST = 0.019, D_est = 0.049). Most importantly, we reveal significant relationships between pairwise F_ST and resistance models created from contemporary land use maps. Specifically, B. vosnesenskii gene flow is most limited by commercial, industrial and transportation‐related impervious cover. Finally, our fine‐scale analysis reveals significant but declining relatedness between individuals at the 1–9 km spatial scale, most likely due to local queen dispersal. Overall, our results indicate that B. vosnesenskii exhibits considerable local dispersal and that regional gene flow is significantly limited by impervious cover associated with urbanization.     

Control of pupal commitment in the imaginal disks of Precis coenia (Lepidoptera: Nymphalidae)

When final (5th) instar larvae of Precis coenia were treated with the juvenile hormone analog (JHA) methoprene, they underwent a supernumerary larval molt, except for certain regions of their imaginal disks, which deposited a normal pupal cuticle. Evidently those regions had already become irreversibly committed to pupal development at the time JHA was applied. By applying JHA at successively later times in the instar, the progression of pupal commitment could be studied. Pupal commitment in the proboscis, antenna, eye, leg and wing imaginal disks occurred in disk-specific patterns. In each imaginal disk there were distinct initiation sites where pupal commitment began during the first few hours of the final larval instar, and from which commitment spread across the remainder of the disk over a 2- to 3-day period. The initiation sites were not always located in homologous regions of the various disks. As a rule, pupal commitment also spread from imaginal disk tissue to surrounding epidermal tissue. The regions of pupal commitment in all disks except those of the wings, coincided with the regions of growth of the disk. Only portions of the disk that had undergone cell division and growth underwent pupal commitment. Shortening the growth period did not prevent pupal commitment in the wing imaginal disk, indicating that, in this disk at least, a normal number of cell divisions was not crucial in reprogramming of disk cells for pupal cuticle synthesis. The apparent growth spurt of imaginal disks that occurs during the last part of the final larval instar is merely the final stage of normal and constant exponential growth. Juvenile hormone (JH) and ecdysteroids appeared to play little role in the regulation of normal imaginal disk growth. Instead, growth of the disks may be under intrinsic control. Interestingly, even though endogenous fluctuation in JH titers do not affect imaginal disk growth, exogenous JHA proved able to inhibit both pupal commitment, cell movement, and growth of the disks during the last larval instar. This function of JH could be important under certain adverse conditions, such as when metamorphosis is delayed in favor of a supernumerary larval molt.     

Root, shoot tissues of Brassica juncea and Cereal secale promote potato health

Brassica species are increasingly being used as cover crops to suppress soil-borne diseases in potato cropping systems. Experiments were conducted in controlled environments and in the field to evaluate the effects of cover crop root or shoot or a combination of root and shoot tissues on potato root and tuber health. In a lab assay we examined the extent to which volatile compounds released from tissues of two cover crop species, rye (Cereale secale L.) and oriental mustard (Brassica juncea L.), could inhibit mycelium growth of two important potato diseases, Rhizoctonia solani and Pythium ultimum. Twenty-four hours into the lab assay, volatile compounds from all residues suppressed fungal growth. After 48 h, marked suppression of hyphal growth continued in the presence of mustard residues but not in the presence of rye tissues or the control without tissues. A 75 L volume container experiment evaluated the effect of incorporating different quantities of mustard shoot and root tissues (none, comparable to field level and fourfold field level) into R. solani and P. ultimum infested soil on potato growth, root health and tuber disease. In the container study, incorporating mustard shoots at the highest dose increased potato yield by 54% and reduced disease rating to 2.3 compared to a severe rating of 4.4 in the control. In the field trial, potato growth, root health and tuber disease levels were evaluated in plots where disease management involved either incorporation of mustard or rye cover crop roots, shoots and whole plants (roots plus shoots) or standard farmer practice of a fumigated fallow as a control. White root tissue was used as a health indicator, and averaged 58 and 78% in the fumigated control and mustard cover crop treatments, respectively. The highest healthy root tissue status (91%) was recorded where whole plants of mustard were incorporated. In contrast to the visual assessment of root and tuber health, tuber yield in the field was not influenced by cover crop treatment. Across experiments, the incorporation of or exposure to whole mustard plants was consistently effective at suppressing soil-borne fungi and promoting healthy roots and tubers, especially at higher rates of biomass. Mustard should be managed so as to maximize incorporated biomass for effective biofumigation. Multipurpose management requiring removal of mustard shoots is incompatible with promoting potato rhizosphere health.     

Characterization of dental pulp stem/stromal cells of Huntington monkey tooth germs

Background

Activation by extracellular ligands of G protein-coupled (GPCRs) and tyrosine kinase receptors (RTKs), results in the generation of second messengers that in turn control specific cell functions. Further, modulation/amplification or inhibition of the initial signalling events, depend on the recruitment onto the plasma membrane of soluble protein effectors. High throughput methodologies to monitor quantitatively second messenger production, have been developed over the last years and are largely used to screen chemical libraries for drug development. On the contrary, no such high throughput methods are yet available for the other aspect of GPCRs regulation, i.e. protein translocation to the plasma membrane, despite the enormous interest of this phenomenon for the modulation of receptor downstream functions. Indeed, to date, the experimental procedures available are either inadequate or complex and expensive.

Results

Here we describe the development of a novel conceptual approach to the study of cytosolic proteins translocation to the inner surface of the plasma membrane. The basis of the technique consists in: i) generating chimeras between the protein of interests and the calcium (Ca²⁺)-sensitive, luminescent photo-protein, aequorin and ii) taking advantage of the large Ca²⁺ concentration [Ca²⁺] difference between bulk cytosolic and the sub-plasma membrane rim.

Conclusion

This approach, that keeps unaffected the translocation properties of the signalling protein, can in principle be applied to any protein that, upon activation, moves from the cytosol to the plasma membrane. Thus, not only the modulation of GPCRs and RTKs can be investigated in this way, but that of all other proteins that can be recruited to the plasma membrane also independently of receptor activation. Moreover, its automated version, which can provide information about the kinetics and concentration-dependence of the process, is also applicable to high throughput screening of drugs affecting the translocation process.     

Reconnecting plants and pollinators: challenges in the restoration of pollination mutualisms

Ecological restoration of plant-pollinator interactions has received surprisingly little attention, despite animal-mediated pollination underpinning reproduction of the majority of higher plants. Here, we offer a conceptual and practical framework for the ecological restoration of pollination mutualisms. Through the use of targeted restoration plantings to attract and sustain pollinators and increased knowledge of the ecological requirements of pollinators, we propose that pollination could be successfully restored in degraded ecosystems. The challenge for pollination biologists is to integrate their findings with those of plant restoration ecologists to ensure sustainable pollination in restored ecosystems.     

Private specificities can dominate the humoral response to self-antigens in patients with cryptogenic fibrosing alveolitis

Background  

The pathogenetic mechanisms that underlie the interstitial lung disease cryptogenic fibrosing alveolitis (CFA) may involve an immunological reaction to unidentified antigens in the lung, resulting in tissue damage.