Monitoring Susceptibility of Western Bean Cutworm (Lepidoptera: Noctuidae) Field Populations to Bacillus thuringiensis Cry1F Protein.

Zea mays L. (maize) hybrids producing the Cry1F protein from Bacillus thuringiensis were first commercialized in the United States in 2003. These products demonstrated varying levels of moderate control, but not immunity to Striacosta albicosta (Smith) (Lepidoptera: Noctuidae) (western bean cutworm). Susceptibility of western bean cutworm to Cry1F protein was assessed in field populations collected in the mid- and western United States in 2003, 2004, 2013, and 2014 using diet bioassay. A meta-analysis of 32 western bean cutworm field collections assessed for susceptibility to Cry1F was conducted to investigate changes in susceptibility over time. Based on meta-analysis results, these data suggest a 5.2-fold increase in median lethal concentration (LC50) response to Cry1F in the 2013­2014 populations compared with collections that were assessed 10 yr earlier. Widespread use of Cry1F-producing maize hybrids over the past 10 yr may have contributed to favoring western bean cutworm populations with tolerance to the Cry1F protein.

Evolution of Resistance by Helicoverpa zea (Lepidoptera: Noctuidae) Infesting Insecticidal Crops in the Southern United States.

We created a deterministic, frequency-based model of the evolution of resistance by corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), to insecticidal traits expressed in crops planted in the heterogeneous landscapes of the southern United States. The model accounts for four generations of selection by insecticidal traits each year. We used the model results to investigate the influence of three factors on insect resistance management (IRM): 1) how does adding a third insecticidal trait to both corn and cotton affect durability of the products, 2) how does unstructured corn refuge influence IRM, and 3) how do block refuges (50% compliance) and blended refuges compare with regard to IRM? When Bt cotton expresses the same number of insecticidal traits, Bt corn with three insecticidal traits provides longer durability than Bt corn with two pyramided traits. Blended refuge provides similar durability for corn products compared with the same level of required block refuge when the rate of refuge compliance by farmers is 50%. Results for Mississippi and Texas are similar, but durabilities for corn traits are surprisingly lower in Georgia, where unstructured corn refuge is the highest of the three states, but refuge for Bt cotton is the lowest of the three states. Thus, unstructured corn refuge can be valuable for IRM but its influence is determined by selection for resistance by Bt cotton.

Snapshot 3D optical coherence tomography system using image mapping spectrometry.

A snapshot 3-Dimensional Optical Coherence Tomography system was developed using Image Mapping Spectrometry. This system can give depth information (Z) at different spatial positions (XY) within one camera integration time to potentially reduce motion artifact and enhance throughput. The current (x,y,λ) datacube of (85×356×117) provides a 3D visualization of sample with 400 µm depth and 13.4 µm in transverse resolution. Axial resolution of 16.0 µm can also be achieved in this proof-of-concept system. We present an analysis of the theoretical constraints which will guide development of future systems with increased imaging depth and improved axial and lateral resolutions.

Assessing the ecological risks from the persistence and spread of feral populations of insect-resistant transgenic maize.

One source of potential harm from the cultivation of transgenic crops is their dispersal, persistence and spread in non-agricultural land. Ecological damage may result from such spread if the abundance of valued species is reduced. The ability of a plant to spread in non-agricultural habitats is called its invasiveness potential. The risks posed by the invasiveness potential of transgenic crops are assessed by comparing in agronomic field trials the phenotypes of the crops with the phenotypes of genetically similar non-transgenic crops known to have low invasiveness potential. If the transgenic and non-transgenic crops are similar in traits believed to control invasiveness potential, it may be concluded that the transgenic crop has low invasiveness potential and poses negligible ecological risk via persistence and spread in non-agricultural habitats. If the phenotype of the transgenic crop is outside the range of the non-transgenic comparators for the traits controlling invasiveness potential, or if the comparative approach is regarded as inadequate for reasons of risk perception or risk communication, experiments that simulate the dispersal of the crop into non-agricultural habitats may be necessary. We describe such an experiment for several commercial insect-resistant transgenic maize events in conditions similar to those found in maize-growing regions of Mexico. As expected from comparative risk assessments, the transgenic maize was found to behave similarly to non-transgenic maize and to be non-invasive. The value of this experiment in assessing and communicating the negligible ecological risk posed by the low invasiveness potential of insect-resistant transgenic maize in Mexico is discussed.

Evolution of gene expression in the Drosophila olfactory system.

Host plant shifts by phytophagous insects play a key role in insect evolution and plant ecology. Such shifts often involve major behavioral changes as the insects must acquire an attraction and/or lose the repulsion to the new host plant's odor and taste. The evolution of chemotactic behavior may be due, in part, to gene expression changes in the peripheral sensory system. To test this hypothesis, we compared gene expression in the olfactory organs of Drosophila sechellia, a narrow ecological specialist that feeds on the fruit of Morinda citrifolia, with its close relatives Drosophila simulans and Drosophila melanogaster, which feed on a wide variety of decaying plant matter. Using whole-genome microarrays and quantitative polymerase chain reaction, we surveyed the entire repertoire of Drosophila odorant receptors (ORs) and odorant-binding proteins (OBPs) expressed in the antennae. We found that the evolution of OR and OBP expression was accelerated in D. sechellia compared both with the genome average in that species and with the rate of OR and OBP evolution in the other species. However, some of the gene expression changes that correlate with D. sechellia's increased sensitivity to Morinda odorants may predate its divergence from D. simulans. Interspecific divergence of olfactory gene expression cannot be fully explained by changes in the relative abundance of different sensilla as some ORs and OBPs have evolved independently of other genes expressed in the same sensilla. A number of OR and OBP genes are upregulated in D. sechellia compared with its generalist relatives. These genes include Or22a, which likely responds to a key odorant of M. citrifolia, and several genes that are yet to be characterized in detail. Increased expression of these genes in D. sechellia may have contributed to the evolution of its unique chemotactic behavior.

Inflammation associated with the postprandial lipolysis of triglyceride-rich lipoproteins by lipoprotein lipase.

Although hypertriglyceridemia has repeatedly been implicated as an atherogenic condition, there are conflicting reports concerning the atherogenicity of products released from triglyceride-rich lipoproteins by lipoprotein lipase. The hydrolysis of triglyceride is a normal process by which chylomicrons and very low-density lipoproteins are metabolized and cleared from the circulation, which would suggest a beneficial role for lipoprotein lipase in reducing circulating levels of triglyceride and, therefore, reducing atherosclerotic burden. However, many in vitro studies have shown that lipolysis products such as fatty acids induce vascular cell inflammation, which can initiate or exacerbate atherosclerosis. This review summarizes the results and implications of recent studies on the effects of lipoprotein lipase on vascular inflammation, expanding upon existing controversy among human postprandial studies, animal models, and in vitro experimental models.

Three-year field monitoring of Cry1F, event DAS-01507-1, maize hybrids for nontarget arthropod effects.

Field studies were conducted over a 3-yr period to investigate the potential effects of cultivating transgenic maize hybrids containing a Cry1F insect-resistant protein on nontarget arthropod abundance. The narrow spectrum of activity of Cry1F against a subset of lepidopteran pest species would not suggest broad-spectrum effects on nontarget arthropods. However, because of the insecticidal nature of Bt proteins, an alternate hypothesis is that some nontargets may be affected by exposure to the protein. To examine this hypothesis at the field level, monitoring for nontarget organism abundance was initiated at four locations across the U.S. Corn Belt from 2004 through 2006. At each location, paired fields (approximately 0.8 ha each) of commercial Cry1F maize hybrids and isogenic nontransgenic control hybrids were planted. Sampling methods used to monitor nontarget organisms included visual surveillance, sticky cards, pitfall traps, and litterbags. Data were analyzed using multivariate analyses to look for a general community level response to the treatments. Analysis of variance was conducted on individual taxa to detect differences distinct from the primary community response. Community level analyses of the nontarget arthropod abundance showed no significant impact on community abundance when comparing Bt with non-Bt maize fields. Analyses of the individual taxa also showed no significant differences in abundance between Bt and non-Bt fields. Results of these studies confirm earlier laboratory testing and support the hypothesis that Cry1F maize does not produce adverse effects on nontarget arthropods occurring in maize fields.

Multiscale optical imaging of rare-earth-doped nanocomposites in a small animal model.

Rare-earth-doped nanocomposites have appealing optical properties for use as biomedical contrast agents, but few systems exist for imaging these materials. We describe the design and characterization of (i) a preclinical system for whole animal in vivo imaging and (ii) an integrated optical coherence tomography/confocal microscopy system for high-resolution imaging of ex vivo tissues. We demonstrate these systems by administering erbium-doped nanocomposites to a murine model of metastatic breast cancer. Short-wave infrared emissions were detected in vivo and in whole organ imaging ex vivo. Visible upconversion emissions and tissue autofluorescence were imaged in biopsy specimens, alongside optical coherence tomography imaging of tissue microstructure. We anticipate that this work will provide guidance for researchers seeking to image these nanomaterials across a wide range of biological models.

Are there age-related differences in learning to optimize speed, accuracy, and energy expenditure?

Studies of age-related differences in manual aiming have indicated that older adults take longer to complete their movements than their younger counterparts because they tend to rely on time-consuming feedback-based control processes. Many authors have suggested that the reliance on feedback is the result of a "play-it-safe" strategy that has been adopted to compensate for a deterioration in accurate and consistent force generation. That is, perhaps because older adults know that their motor systems are not as reliable as the systems were at a younger age, they plan shorter movements that conserve time and space for feedback control to correct their programmed actions. The vast majority of the previous studies that have revealed these age-related differences in aiming, however, have used computer-based tasks that involve the transformation of perceptual into motor space. In the present experiment, older and younger adults completed real aiming movements over three sessions. The results suggest that, when acting in a real environment, the main difference between older and younger adults in movement execution lies in the efficient use of response-related feedback, not in the programming of movement.

Seeing vs. believing: Is believing sufficient to activate the processes of response co-representation?

It has been suggested that the observation of another person's action affects the behavior of the observer because the observation of action leads to the excitation of similar response codes in the observer. It is unknown, however, if one must witness the action or if it is sufficient for one to believe that the other agent is responding for response co-representation to occur. To this end, participants in the present study performed a joint spatial-compatibility task with a confederate when: (1) the confederate sat beside the participant; and, (2) the confederate left the room and told the participant that they would continue to perform their component of the task on a networked computer in another room. Even though participants believed that the confederate performed the task in another room, joint spatial-compatibility was only observed when the confederate was present. These results reveal that the actions of another person may only be represented by the observer when the observer is able to witness a portion of the action.

Surveillance nanotechnology for multi-organ cancer metastases.

The identification and molecular profiling of early metastases remains a major challenge in cancer diagnostics and therapy. Most in vivo imaging methods fail to detect small cancerous lesions, a problem that is compounded by the distinct physical and biological barriers associated with different metastatic niches. Here, we show that intravenously injected rare-earth-doped albumin-encapsulated nanoparticles emitting short-wave infrared light (SWIR) can detect targeted metastatic lesions in vivo, allowing for the longitudinal tracking of multi-organ metastases. In a murine model of basal human breast cancer, the nanoprobes enabled whole-body SWIR detection of adrenal gland microlesions and bone lesions that were undetectable via contrast-enhanced magnetic resonance imaging (CE-MRI) as early as, respectively, three weeks and five weeks post-inoculation. Whole-body SWIR imaging of nanoprobes functionalized to differentially target distinct metastatic sites and administered to a biomimetic murine model of human breast cancer resolved multi-organ metastases that showed varied molecular profiles at the lungs, adrenal glands and bones. Real-time surveillance of lesions in multiple organs should facilitate pre-therapy and post-therapy monitoring in preclinical settings.

Quantitative genetics of natural variation of behavior in Drosophila melanogaster: the possible role of the social environment on creating persistent patterns of group activity.

Using a set of nine effectively isogenic lines collected from nature in 1998, we observed unperturbed behaviors of mixed-sex groups of Drosophila melanogaster. We repeatedly scanned replicated groups of genetically identical individuals, five females and five males, and recorded the behavior of each individual (i.e., walking, feeding, grooming, flying, courting, mating, fighting, or resting). From these behaviors, we made a composite variable of activity for our quantitative genetic analysis. Genotypes differed in activity, explaining 14.41% of the variation in activity; 8.60% of the variation was explained by a significant genotype x sex interaction, which signifies genetic variation for sexual dimorphism in behavior. Phenotypic plasticity explained 11.13% of the variation in activity. Different genotypes and sexes within genotypes had different rank orders of the component behaviors that contribute to activity. We found no effect of common rearing environment. Instead, differences between replicate groups within genotype accounted for 19.47% variation in activity, and activity was significantly repeatable across scans. This emergent group behavior is likely caused by differences between groups of interacting individuals, even though individuals were genetically identical across groups. Thus, emergent group behavior explained almost as much variation in activity as the combined sources of genetic variation (23.01%), and this is an additional level on which selection could operate: individuals and groups. We discuss how differences among groups could change patterns of additive genetic variation available for evolution. Furthermore, because the behavior of an individual is influenced by conspecifics, genotype interactions between individuals could contribute to indirect selection. Finally, if we consider activity as a syndrome governing all component behaviors with strong genetic correlations among behaviors within an individual, then these component behaviors cannot evolve independently. These results suggest that reductionist approaches of molecular behavior genetics may be incomplete and/or misleading when considering similar phenotypes at the population level or when trying to understand how behaviors evolve.

Line-scanning confocal microscopy for high-resolution imaging of upconverting rare-earth-based contrast agents.

Rare-earth (RE) doped nanocomposites emit visible luminescence when illuminated with continuous wave near-infrared light, making them appealing candidates for use as contrast agents in biomedical imaging. However, the emission lifetime of these materials is much longer than the pixel dwell times used in scanning intravital microscopy. To overcome this limitation, we have developed a line-scanning confocal microscope for high-resolution, optically sectioned imaging of samples labeled with RE-based nanomaterials. Instrument performance is quantified using calibrated test objects. NaYF4 : Er,Yb nanocomposites are imaged in vitro, and in ex vivo tissue specimens, with direct comparison to point-scanning confocal microscopy. We demonstrate that the extended pixel dwell time of line-scanning confocal microscopy enables subcellular-level imaging of these nanomaterials while maintaining optical sectioning. The line-scanning approach thus enables microscopic imaging of this emerging class of contrast agents for preclinical studies, with the potential to be adapted for real-time in vivo imaging in the clinic.

Preliminary estimates of effects of a cognitive behavioral treatment system on drop-out rates in exercise programs at three YMCA facilities.

The effect of a treatment system based on principles of social cognitive and cognitive behavioral theory was estimated through case studies conducted in three YMCA fitness facilities. Significant (p < .05) reductions in drop-out rates of 23% (Georgia sample; n = 171), 27% (California sample; n = 159), and 21% (Florida sample; = 83) were found for the new and returning exercisers over the 5- and 6-mo. study time frames. Age was not related to drop-out rates in the treatment groups. Findings were consistent with previous research on earlier versions of the present treatment system, conducted with larger sample sizes within different fitness and wellness venues in the USA, U.K., and Italy. Replication, more detailed assessment of outputs of exercise, and tests of further revisions of the protocol were suggested to extend this preliminary research.

Design and characterization of a handheld multimodal imaging device for the assessment of oral epithelial lesions.

A compact handpiece combining high resolution fluorescence (HRF) imaging with optical coherence tomography (OCT) was developed to provide real-time assessment of oral lesions. This multimodal imaging device simultaneously captures coregistered en face images with subcellular detail alongside cross-sectional images of tissue microstructure. The HRF imaging acquires a 712 × 594 µm² field-of-view at the sample with a spatial resolution of 3.5 µm. The OCT images were acquired to a depth of 1.5 mm with axial and lateral resolutions of 9.3 and 8.0 µm, respectively. HRF and OCT images are simultaneously displayed at 25 fps. The handheld device was used to image a healthy volunteer, demonstrating the potential for in vivo assessment of the epithelial surface for dysplastic and neoplastic changes at the cellular level, while simultaneously evaluating submucosal involvement. We anticipate potential applications in real-time assessment of oral lesions for improved surveillance and surgical guidance.

Effectiveness of a steam cleaning unit for disinfection in a veterinary hospital.

OBJECTIVE: To evaluate whether the application of steam to a variety of surface types in a veterinary hospital would effectively reduce the number of bacteria. SAMPLE: 5 surface types. PROCEDURES: Steam was applied as a surface treatment for disinfection to 18 test sites of 5 surface types in a veterinary hospital. A pretreatment sample was obtained by collection of a swab specimen from the left side of each defined test surface. Steam disinfection was performed on the right side of each test surface, and a posttreatment sample was then collected in the same manner from the treated (right) side of each test surface. Total bacteria for pretreatment and posttreatment samples were quantified by heterotrophic plate counts and for Staphylococcus aureus, Pseudomonas spp, and total coliforms by counts on selective media. RESULTS: Significant reductions were observed in heterotrophic plate counts after steam application to dog runs and dog kennel floors. A significant reduction in counts of Pseudomonas spp was observed after steam application to tub sinks. Bacterial counts were reduced, but not significantly, on most other test surfaces that had adequate pretreatment counts for quantification. CONCLUSIONS AND CLINICAL RELEVANCE: Development of health-care-associated infections is of increasing concern in human and veterinary medicine. The application of steam significantly reduced bacterial numbers on a variety of surfaces within a veterinary facility. Steam disinfection may prove to be an alternative or adjunct to chemical disinfection within veterinary practices.

Characterization of thin poly(dimethylsiloxane)-based tissue-simulating phantoms with tunable reduced scattering and absorption coefficients at visible and near-infrared wavelengths.

Optical phantoms are used in the development of various imaging systems. For certain applications, the development of thin phantoms that simulate the physical size and optical properties of tissue is important. Here, we demonstrate a method for producing thin phantom layers with tunable optical properties using poly(dimethylsiloxane) (PDMS) as a substrate material. The thickness of each layer (between 115 and 880 µm) was controlled using a spin coater. The reduced scattering and absorption coefficients were controlled using titanium dioxide and alcohol-soluble nigrosin, respectively. These optical coefficients were quantified at six discrete wavelengths (591, 631, 659, 691, 731, and 851 nm) at varying concentrations of titanium dioxide and nigrosin using spatial frequency domain imaging. From the presented data, we provide lookup tables to determine the appropriate concentrations of scattering and absorbing agents to be used in the design of PDMS-based phantoms with specific optical coefficients. In addition, heterogeneous phantoms mimicking the layered features of certain tissue types may be fabricated from multiple stacked layers, each with custom optical properties. These thin, tunable PDMS optical phantoms can simulate many tissue types and have broad imaging calibration applications in endoscopy, diffuse optical spectroscopic imaging, and optical coherence tomography, etc.

Factors that affect action possibility judgements: recent experience with the action and the current body state.

It has been suggested that action possibility judgements are formed through a covert simulation of the to-be-executed action. We sought to determine whether the motor system (via a common coding mechanism) influences this simulation, by investigating whether action possibility judgements are influenced by experience with the movement task (Experiments 1 and 2) and current body states (Experiment 3). The judgement task in each experiment involved judging whether it was possible for a person's hand to accurately move between two targets at presented speeds. In Experiment 1, participants completed the action judgements before and after executing the movement they were required to judge. Results were that judged movement times after execution were closer to the actual execution time than those prior to execution. The results of Experiment 2 suggest that the effects of execution on judgements were not due to motor activation or perceptual task experience-alternative explanations of the execution-mediated judgement effects. Experiment 3 examined how judged movement times were influenced by participants wearing weights. Results revealed that wearing weights increased judged movement times. These results suggest that the simulation underlying the judgement process is connected to the motor system, and that simulations are dynamically generated, taking into account recent experience and current body state.

Innate immune responses of Drosophila melanogaster are altered by spaceflight.

Alterations and impairment of immune responses in humans present a health risk for space exploration missions. The molecular mechanisms underpinning innate immune defense can be confounded by the complexity of the acquired immune system of humans. Drosophila (fruit fly) innate immunity is simpler, and shares many similarities with human innate immunity at the level of molecular and genetic pathways. The goals of this study were to elucidate fundamental immune processes in Drosophila affected by spaceflight and to measure host-pathogen responses post-flight. Five containers, each containing ten female and five male fruit flies, were housed and bred on the space shuttle (average orbit altitude of 330.35 km) for 12 days and 18.5 hours. A new generation of flies was reared in microgravity. In larvae, the immune system was examined by analyzing plasmatocyte number and activity in culture. In adults, the induced immune responses were analyzed by bacterial clearance and quantitative real-time polymerase chain reaction (qPCR) of selected genes following infection with E. coli. The RNA levels of relevant immune pathway genes were determined in both larvae and adults by microarray analysis. The ability of larval plasmatocytes to phagocytose E. coli in culture was attenuated following spaceflight, and in parallel, the expression of genes involved in cell maturation was downregulated. In addition, the level of constitutive expression of pattern recognition receptors and opsonins that specifically recognize bacteria, and of lysozymes, antimicrobial peptide (AMP) pathway and immune stress genes, hallmarks of humoral immunity, were also reduced in larvae. In adults, the efficiency of bacterial clearance measured in vivo following a systemic infection with E. coli post-flight, remained robust. We show that spaceflight altered both cellular and humoral immune responses in Drosophila and that the disruption occurs at multiple interacting pathways.

Conidiation color mutants of Aspergillus fumigatus are highly pathogenic to the heterologous insect host Galleria mellonella.

The greater wax moth Galleria mellonella has been widely used as a heterologous host for a number of fungal pathogens including Candida albicans and Cryptococcus neoformans. A positive correlation in pathogenicity of these yeasts in this insect model and animal models has been observed. However, very few studies have evaluated the possibility of applying this heterologous insect model to investigate virulence traits of the filamentous fungal pathogen Aspergillus fumigatus, the leading cause of invasive aspergillosis. Here, we have examined the impact of mutations in genes involved in melanin biosynthesis on the pathogenicity of A. fumigatus in the G. mellonella model. Melanization in A. fumigatus confers bluish-grey color to conidia and is a known virulence factor in mammal models. Surprisingly, conidial color mutants in B5233 background that have deletions in the defined six-gene cluster required for DHN-melanin biosynthesis caused enhanced insect mortality compared to the parent strain. To further examine and confirm the relationship between melanization defects and enhanced virulence in the wax moth model, we performed random insertional mutagenesis in the Af293 genetic background to isolate mutants producing altered conidia colors. Strains producing conidia of previously identified colors and of novel colors were isolated. Interestingly, these color mutants displayed a higher level of pathogenicity in the insect model compared to the wild type. Although some of the more virulent color mutants showed increased resistance to hydrogen peroxide, overall phenotypic characterizations including secondary metabolite production, metalloproteinase activity, and germination rate did not reveal a general mechanism accountable for the enhanced virulence of these color mutants observed in the insect model. Our observations indicate instead, that exacerbated immune response of the wax moth induced by increased exposure of PAMPs (pathogen-associated molecular patterns) may cause self-damage that results in increased mortality of larvae infected with the color mutants. The current study underscores the limitations of using this insect model for inferring the pathogenic potential of A. fumigatus strains in mammals, but also points to the importance of understanding the innate immunity of the insect host in providing insights into the pathogenicity level of different fungal strains in this model. Additionally, our observations that melanization defective color mutants demonstrate increased virulence in the insect wax moth, suggest the potential of using melanization defective mutants of native insect fungal pathogens in the biological control of insect populations.