Examining disruption of pheromone communication in Choristoneura rosaceana and Pandemis limitata using microencapsulated (Z)-11-tetradecenyl acetate applied in a laboratory flight tunnel

Pheromone‐based mating disruption of lepidopteran pests (Tortricidae) of pome fruits using hand‐applied dispensing systems has become standard management practice for many producers in western North America. Sprayable microencapsulated (MEC) pheromone formulations that enable the application of pheromone controls with other orchard sprays and assist in the development of multispecies mating‐disruption systems are currently under development. Responses of male Choristoneura rosaceana (Harris) and Pandemis limitata (Robinson) (Lepidoptera: Tortricidae) to calling females in clean air, and air treated with their major pheromone component (Z)‐11‐tetradecenyl acetate (Z11‐14:OAc), released from a 3M sprayable pheromone formulation containing proprietary 3M Phase I microcapsules, applied at doses of 1, 10, and 100 mg of active ingredient (ai) m^?2 to the upwind end of a flight tunnel (equivalent to field rates of 10, 100, and 1000 g ai ha^?1) were compared in laboratory flight tunnels. In both species, disorientation was found to be dose‐dependent, because relative to male orientation to calling females in clean air, the orientation of male P. limitata was disrupted 23.3, 46.3, and 71.3%, and orientation by male C. rosaceana was disrupted 31.6, 37.7, and 45.8% by treatment doses of 1, 10, and 100 mg m^?2, respectively. Latency of male responses to calling females in a background of Z11‐14:OAc relative to responses in clean air was also dose‐dependent. Albeit short, the disruption lasted 26, 74, and 218 h in P. limitata and 30, 54, and 174 h in C. rosaceana at each application rate, respectively. Disruption by pheromone treatment was greater in P. limitata than in C. rosaceana. This difference may be correlated with species’ differences in the pheromone release rates of females. Mechanisms of disruption invoked by this 3M MEC pheromone formulation are discussed in relation to issues of its longevity and observed differences in the effects against the two species. It appears possible to evaluate relative activity of MEC pheromones in a laboratory setting which may aid in development of new formulations for mating disruption.     

Muscle palmitate uptake and binding are saturable and inhibited by antibodies to FABPPM

Studies show that uptake of long-chain fatty acids (LCFA) across the plasma membranes (PM) may occur partly via a carrier-mediated process and that the plasma membrane fatty acid-binding protein (FABP_PM) may be a component of this system. To test the hypothesis that FABP_PM is involved in transsarcolemmal transport of LCFA in muscle, we measured palmitate uptake in giant sarcolemmal vesicles and palmitate binding to PM proteins in rat muscles, (1) in the presence of increasing amounts of unbound palmitate and (2) in the absence or presence of antibody to FABP_PM. Both palmitate uptake and binding were found to be saturable functions of the unbound palmitate concentration with calculated V_max values of 10.5 ± 1.2 pmol/mg protein/15 sec and 45.6 ± 2.9 nmol/mg protein/15 min and K_m values of 12.8 ± 3.8 and 18.4 ± 1.8 nmol/L, respectively. The V_max values for both palmitate uptake and binding were significantly decreased by 75-79% in the presence of a polyclonal antibody to the rat hepatic FABP_PM. Antibody inhibition was found to be dose-dependent and specific to LCFA. Glucose uptake was not affected by the presence of the antibody to FABP_PM. Palmitate uptake and binding were also inhibited in the presence of trypsin and phloretin. These results support the hypothesis that transsarcolemmal LCFA transport occurs in part by a carrier-mediated process and that FABP_PM is a component of this process in muscle.     

Safe From Harm: Learned,Instructed, and Symbolic Generalization Pathways of Human Threat-Avoidance

Avoidance of threatening or unpleasant events is usually an adaptive behavioural strategy. Sometimes, however, avoidance can become chronic and lead to impaired daily functioning. Excessive threat-avoidance is a central diagnostic feature of anxiety disorders, yet little is known about whether avoidance acquired in the absence of a direct history of conditioning with a fearful event differs from directly learned avoidance. In the present study, we tested whether avoidance acquired indirectly via verbal instructions and symbolic generalization result in similar levels of avoidance behaviour and threat-beliefs to avoidance acquired after direct learning. Following fear conditioning in which one conditioned stimulus was paired with shock (CS+) and another was not (CS−), participants either learned or were instructed to make a response that cancelled impending shock. Three groups were then tested with a learned CS+ and CS− (learned group), instructed CS+ (instructed group), and generalized CS+ (derived group) presentations. Results showed similar levels of avoidance behaviour and threat-belief ratings about the likelihood of shock across each of the three pathways despite the different mechanisms by which they were acquired. Findings have implications for understanding the aetiology of clinical avoidance in anxiety.     

Characterisation of Ilomastat for Prolonged Ocular Drug Release

We are developing tablet dosage forms for implantation directly into the subconjunctival space of the eye. The matrix metalloproteinase inhibitor, ilomastat, has previously been shown to be efficacious at suppressing scarring following glaucoma filtration surgery (GFS). We report on the physical characterisation of ilomastat which is being developed for ocular implantation. Since ilomastat is being considered for implantation it is necessary to examine its polymorphs and their influence on aspects of the in vitro drug release profile. X-ray powder diffraction identified two polymorphs of ilomastat from different commercial batches of the compound. Tablets were prepared from the two different polymorphs. Isothermal perfusion calorimetry was used to show that amorphous content is not increased during tablet formulation. The melting points of the two polymorphs are 188 and 208°C as determined by differential scanning calorimetry. Utilising single crystal X-ray diffraction, the structural conformations and packing arrangements of the different polymorphs were determined. The orthorhombic crystal crystallised as a monohydrate while the second monoclinic crystal form is non-solvated. Ilomastat tablets prepared from the two different solid forms exhibited similar drug release profiles in vitro under conditions mimicking the aqueous composition, volume and flow of the subconjunctival space after GFS. This suggests that a reproducible dose at each time point during release after implantation should be achievable in vivo with ilomastat tablets prepared from the two polymorphs identified.     

Interspecific mate choice by late-courting male western grebes

In mixed populations of western and Clark's grebes, advertisingcalls by males and females play a critical role in mate choiceand reproductive isolation. We conducted field playback experimentsthat tested whether courting western grebe males became lesschoosy in their responses to female Clark's grebe calls as themating season progressed and mating opportunities diminished.Late-courting western grebe males were much more likely to answerand approach advertising calls of Dark's grebe females thanwere males courting earlier in the season. This change in responsivenessoccurred as the operational sex ratio index of the populationapproached 3:1 male calls per female call. These and field censusdata support the hypothesis that late-season hybridization betweenthese two closely related species may be a result not of speciesmisidentification, but of active and adaptive mate choice byindividuals with limited alternatives.     

Meta‐analysis suggests negative,but pCO2‐specific,effects of ocean acidification on the structural and functional properties of crustacean biomaterials

Crustaceans comprise an ecologically and morphologically diverse taxonomic group. They are typically considered resilient to many environmental perturbations found in marine and coastal environments, due to effective physiological regulation of ions and hemolymph pH, and a robust exoskeleton. Ocean acidification can affect the ability of marine calcifying organisms to build and maintain mineralized tissue and poses a threat for all marine calcifying taxa. Currently, there is no consensus on how ocean acidification will alter the ecologically relevant exoskeletal properties of crustaceans. Here, we present a systematic review and meta‐analysis on the effects of ocean acidification on the crustacean exoskeleton, assessing both exoskeletal ion content (calcium and magnesium) and functional properties (biomechanical resistance and cuticle thickness). Our results suggest that the effect of ocean acidification on crustacean exoskeletal properties varies based upon seawater pCO₂ and species identity, with significant levels of heterogeneity for all analyses. Calcium and magnesium content was significantly lower in animals held at pCO₂ levels of 1500–1999 µatm as compared with those under ambient pCO₂. At lower pCO₂ levels, however, statistically significant relationships between changes in calcium and magnesium content within the same experiment were observed as follows: a negative relationship between calcium and magnesium content at pCO₂ of 500–999 µatm and a positive relationship at 1000–1499 µatm. Exoskeleton biomechanics, such as resistance to deformation (microhardness) and shell strength, also significantly decreased under pCO₂ regimes of 500–999 µatm and 1500–1999 µatm, indicating functional exoskeletal change coincident with decreases in calcification. Overall, these results suggest that the crustacean exoskeleton can be susceptible to ocean acidification at the biomechanical level, potentially predicated by changes in ion content, when exposed to high influxes of CO₂. Future studies need to accommodate the high variability of crustacean responses to ocean acidification, and ecologically relevant ranges of pCO₂ conditions, when designing experiments with conservation‐level endpoints.     

Canonical correlation analysis of marine macrobenthos survey data

Canonical correlation analysis is applied to measurements of environmental variables and species distributions made during a survey of macrobenthos around a sewage-treatment farm drain. The implications of data reduction, necessary to enable the method to proceed, are discussed. The amount of data was reduced by discarding the rarest species, discarding species occurring at fewest stations, and including only those species and environmental variables which correlated highly with the greatest number of other variables. Only the third data-reduction scheme gave ecologically sensible results. Use of station scores on the first two canonical variates (CV1 and CV2) enabled the sampling grid to be divided into a group of nearshore stations, a group of intermediate depth, and a group of deep offshore stations. Loadings of environmental variables on the canonical variates were found to be unstable but correlations between these variables and canonical variates enabled the variates to be interpreted: CV1 as a gradient of depth and associated changes in sediment characteristics, CV2 with depth- and nutrient-related components, and CV3 as patchiness in sediment characteristics different from that normally expected with depth. Use of correlations between species and canonical variates enables definition of two major species groups, one confined to nearshore environments and a second offshore. These groups (and their sub-groups) related well to groups defined previously by hierarchical classification. It is concluded that, with careful attention to the method of data reduction, canonical correlation analysis can be an effective tool in the analysis of marine benthic survey data.     

The USDA Barley Core Collection: Genetic Diversity,Population Structure,and Potential for Genome-Wide Association Studies

New sources of genetic diversity must be incorporated into plant breeding programs if they are to continue increasing grain yield and quality, and tolerance to abiotic and biotic stresses. Germplasm collections provide a source of genetic and phenotypic diversity, but characterization of these resources is required to increase their utility for breeding programs. We used a barley SNP iSelect platform with 7,842 SNPs to genotype 2,417 barley accessions sampled from the USDA National Small Grains Collection of 33,176 accessions. Most of the accessions in this core collection are categorized as landraces or cultivars/breeding lines and were obtained from more than 100 countries. Both STRUCTURE and principal component analysis identified five major subpopulations within the core collection, mainly differentiated by geographical origin and spike row number (an inflorescence architecture trait). Different patterns of linkage disequilibrium (LD) were found across the barley genome and many regions of high LD contained traits involved in domestication and breeding selection. The genotype data were used to define ‘mini-core’ sets of accessions capturing the majority of the allelic diversity present in the core collection. These ‘mini-core’ sets can be used for evaluating traits that are difficult or expensive to score. Genome-wide association studies (GWAS) of ‘hull cover’, ‘spike row number’, and ‘heading date’ demonstrate the utility of the core collection for locating genetic factors determining important phenotypes. The GWAS results were referenced to a new barley consensus map containing 5,665 SNPs. Our results demonstrate that GWAS and high-density SNP genotyping are effective tools for plant breeders interested in accessing genetic diversity in large germplasm collections.     

Trisubstituted and tetrasubstituted pyrazolines as a novel class of cell-growth inhibitors in tumor cells with wild type p53

Derivatives with scaffolds of 1,3,5-tri-substituted pyrazoline and 1,3,4,5-tetra-substituted pyrazoline were synthesized and tested for their inhibitory effects versus the p53^+/+ HCT116 and p53^?/? H1299 human tumor cell lines. Several compounds were active against the two cell lines displaying IC₅₀ values in the low micromolar range with a clearly more pronounced effect on the p53^+/+ HCT116 cells. The compound class shows excellent developability due to the modular synthesis, allowing independent optimization of all three to four key substituents to improve the properties of the molecules.     

CLASI-FISH: Principles of combinatorial labeling and spectral imaging

Although the number of phylotypes present in a microbial community may number in the hundreds or more, until recently, fluorescence in situ hybridization has been used to label, at most, only a handful of different phylotypes in a single sample. We recently developed a technique, CLASI-FISH for combinatorial labeling and spectral imaging – fluorescence in situ hybridization, to greatly expand the number of distinguishable taxa in a single FISH experiment. The CLASI technique involves labeling microbes of interest with combinations of probes coupled with spectral imaging to allow the use of fluorophores with highly overlapping excitation and emission spectra. Here, we present the basic principles and theory of CLASI-FISH along with some guidelines for performing CLASI-FISH experiments. We further include a protocol for creating fluorescence spectral reference standards, a vital component of successful CLASI-FISH.