The utilization of Blaptica dubia cockroaches as an in vivo model to test antibiotic efficacy.

Insects are now well recognized as biologically relevant alternative hosts for dozens of mammalian pathogens and they are routinely used in microbial pathogenesis studies. Unfortunately, these models have yet to be incorporated into the drug development pipeline. The purpose of this work was to begin to evaluate the utility of orange spotted (Blaptica dubia) cockroaches in early antibiotic characterization. To determine whether these model hosts could exhibit mortality when infected with bacteria that are pathogenic to humans, we subjected B. dubia roaches to a range of infectious doses of Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, and Acinetobacter baumannii to identify the medial lethal dose. These results showed that lethal disease did not develop following infection of high doses of S. aureus, and A. baumannii. However, cockroaches infected with E. coli and K. pneumoniae succumbed to infection (LD50s of 5.82 × 106 and 2.58 × 106 respectively) suggesting that this model may have limitations based on pathogen specificity. However, because these cockroaches were susceptible to infection from E. coli and K. pneumoniae, we used these bacterial strains for subsequent antibiotic characterization studies. These studies suggested that ß-lactam antibiotic persistence and dose was associated with reduction of hemolymph bacterial burden. Moreover, our data indicated that the reduction of bacterial CFU was directly due to the drug activity. Altogether, this work suggests that the orange-spotted cockroach infection model provides an alternative in vivo setting from which antibiotic efficacy can be evaluated.

Exemplar learning reveals the representational origins of expert category perception.

Irrespective of whether one has substantial perceptual expertise for a class of stimuli, an observer invariably encounters novel exemplars from this class. To understand how novel exemplars are represented, we examined the extent to which previous experience with a category constrains the acquisition and nature of representation of subsequent exemplars from that category. Participants completed a perceptual training paradigm with either novel other-race faces (category of experience) or novel computer-generated objects (YUFOs) that included pairwise similarity ratings at the beginning, middle, and end of training, and a 20-d visual search training task on a subset of category exemplars. Analyses of pairwise similarity ratings revealed multiple dissociations between the representational spaces for those learning faces and those learning YUFOs. First, representational distance changes were more selective for faces than YUFOs; trained faces exhibited greater magnitude in representational distance change relative to untrained faces, whereas this trained-untrained distance change was much smaller for YUFOs. Second, there was a difference in where the representational distance changes were observed; for faces, representations that were closer together before training exhibited a greater distance change relative to those that were farther apart before training. For YUFOs, however, the distance changes occurred more uniformly across representational space. Last, there was a decrease in dimensionality of the representational space after training on YUFOs, but not after training on faces. Together, these findings demonstrate how previous category experience governs representational patterns of exemplar learning as well as the underlying dimensionality of the representational space.

Blowing epithelial cell bubbles with GumB: ShlA-family pore-forming toxins induce blebbing and rapid cellular death in corneal epithelial cells.

Medical devices, such as contact lenses, bring bacteria in direct contact with human cells. Consequences of these host-pathogen interactions include the alteration of mammalian cell surface architecture and induction of cellular death that renders tissues more susceptible to infection. Gram-negative bacteria known to induce cellular blebbing by mammalian cells, Pseudomonas and Vibrio species, do so through a type III secretion system-dependent mechanism. This study demonstrates that a subset of bacteria from the Enterobacteriaceae bacterial family induce cellular death and membrane blebs in a variety of cell types via a type V secretion-system dependent mechanism. Here, we report that ShlA-family cytolysins from Proteus mirabilis and Serratia marcescens were required to induce membrane blebbling and cell death. Blebbing and cellular death were blocked by an antioxidant and RIP-1 and MLKL inhibitors, implicating necroptosis in the observed phenotypes. Additional genetic studies determined that an IgaA family stress-response protein, GumB, was necessary to induce blebs. Data supported a model where GumB and shlBA are in a regulatory circuit through the Rcs stress response phosphorelay system required for bleb formation and pathogenesis in an invertebrate model of infection and proliferation in a phagocytic cell line. This study introduces GumB as a regulator of S. marcescens host-pathogen interactions and demonstrates a common type V secretion system-dependent mechanism by which bacteria elicit surface morphological changes on mammalian cells. This type V secretion-system mechanism likely contributes bacterial damage to the corneal epithelial layer, and enables access to deeper parts of the tissue that are more susceptible to infection.

Temporal Dynamics of Shape Processing Differentiate Contributions of Dorsal and Ventral Visual Pathways.

Although shape perception is primarily considered a function of the ventral visual pathway, previous research has shown that both dorsal and ventral pathways represent shape information. Here, we examine whether the shape-selective electrophysiological signals observed in dorsal cortex are a product of the connectivity to ventral cortex or are independently computed. We conducted multiple EEG studies in which we manipulated the input parameters of the stimuli so as to bias processing to either the dorsal or ventral visual pathway. Participants viewed displays of common objects with shape information parametrically degraded across five levels. We measured shape sensitivity by regressing the amplitude of the evoked signal against the degree of stimulus scrambling. Experiment 1, which included grayscale versions of the stimuli, served as a benchmark establishing the temporal pattern of shape processing during typical object perception. These stimuli evoked broad and sustained patterns of shape sensitivity beginning as early as 50 msec after stimulus onset. In Experiments 2 and 3, we calibrated the stimuli such that visual information was delivered primarily through parvocellular inputs, which mainly project to the ventral pathway, or through koniocellular inputs, which mainly project to the dorsal pathway. In the second and third experiments, shape sensitivity was observed, but in distinct spatio-temporal configurations from each other and from that elicited by grayscale inputs. Of particular interest, in the koniocellular condition, shape selectivity emerged earlier than in the parvocellular condition. These findings support the conclusion of distinct dorsal pathway computations of object shape, independent from the ventral pathway.

Distinct neural processes for the perception of familiar versus unfamiliar faces along the visual hierarchy revealed by EEG.

Humans recognize faces with ease, despite the complexity of the task and of the visual system which underlies it. Different spatial regions, including both the core and extended face processing networks, and distinct temporal stages of processing have been implicated in face recognition, but there is ongoing controversy regarding the extent to which the mechanisms for recognizing a familiar face differ from those for an unfamiliar face. Here, we used electroencephalogram (EEG) and flicker SSVEP, a high signal-to-noise approach, and searchlight decoding methods to elucidate the mechanisms mediating the processing of familiar and unfamiliar faces in the time domain. Familiar and unfamiliar faces were presented periodically at 15 Hz, 6 Hz and 3.75 Hz either upright or inverted in separate blocks, with the rationale that faster frequencies require shorter processing times per image and tap into fundamentally different levels of visual processing. The 15 Hz trials, likely to reflect early visual processing, exhibited enhanced neural responses for familiar over unfamiliar face trials, but only when the faces were upright. In contrast, decoding methods revealed similar classification accuracies for upright and inverted faces for both familiar and unfamiliar faces. For the 6 Hz frequency, familiar faces had lower amplitude responses than unfamiliar faces, and decoding familiarity was more accurate for upright compared with inverted faces. Finally, the 3.75 Hz frequency revealed no main effects of familiarity, but decoding showed significant correlations with behavioral ratings of face familiarity, suggesting that activity evoked by this slow presentation frequency reflected higher-level, cognitive aspects of familiarity processing. This three-way dissociation between frequencies reveals that fundamentally different stages of the visual hierarchy are modulated by face familiarity. The combination of experimental and analytical approaches used here represent a novel method for elucidating spatio-temporal characteristics within the visual system.

Numerosity representation is encoded in human subcortex.

Certain numerical abilities appear to be relatively ubiquitous in the animal kingdom, including the ability to recognize and differentiate relative quantities. This skill is present in human adults and children, as well as in nonhuman primates and, perhaps surprisingly, is also demonstrated by lower species such as mosquitofish and spiders, despite the absence of cortical computation available to primates. This ubiquity of numerical competence suggests that representations that connect to numerical tasks are likely subserved by evolutionarily conserved regions of the nervous system. Here, we test the hypothesis that the evaluation of relative numerical quantities is subserved by lower-order brain structures in humans. Using a monocular/dichoptic paradigm, across four experiments, we show that the discrimination of displays, consisting of both large (5-80) and small (1-4) numbers of dots, is facilitated in the monocular, subcortical portions of the visual system. This is only the case, however, when observers evaluate larger ratios of 3:1 or 4:1, but not smaller ratios, closer to 1:1. This profile of competence matches closely the skill with which newborn infants and other species can discriminate numerical quantity. These findings suggest conservation of ontogenetically and phylogenetically lower-order systems in adults' numerical abilities. The involvement of subcortical structures in representing numerical quantities provokes a reconsideration of current theories of the neural basis of numerical cognition, inasmuch as it bolsters the cross-species continuity of the biological system for numerical abilities.

The orange spotted cockroach (Blaptica dubia, Serville 1839) is a permissive experimental host for Francisella tularensis.

Francisella tularensis is a zoonotic bacterial pathogen that causes severe disease in a wide range of host animals, including humans. Well-developed murine models of F. tularensis pathogenesis are available, but they do not meet the needs of all investigators. However, researchers are increasingly turning to insect host systems as a cost-effective alternative that allows greater increased experimental throughput without the regulatory requirements associated with the use of mammals in biomedical research. Unfortunately, the utility of previously-described insect hosts is limited because of temperature restriction, short lifespans, and concerns about the immunological status of insects mass-produced for other purposes. Here, we present a novel host species, the orange spotted (OS) cockroach (Blaptica dubia), that overcomes these limitations and is readily infected by F. tularensis. Intrahemocoel inoculation was accomplished using standard laboratory equipment and lethality was directly proportional to the number of bacteria injected. Progression of infection differed in insects housed at low and high temperatures and F. tularensis mutants lacking key virulence components were attenuated in OS cockroaches. Finally, antibiotics were delivered to infected OS cockroaches by systemic injection and controlled feeding; in the latter case, protection correlated with oral bioavailability in mammals. Collectively, these results demonstrate that this new host system provides investigators with a new tool capable of interrogating F. tularensis virulence and immune evasion in situations where mammalian models are not available or appropriate, such as undirected screens of large mutant libraries.

Hemispheric Organization in Disorders of Development.

A recent theoretical account posits that, during the acquisition of word recognition in childhood, the pressure to couple visual and language representations in the left hemisphere (LH) results in competition with the LH representation of faces, which consequently become largely, albeit not exclusively, lateralized to the right hemisphere (RH). We explore predictions from this hypothesis using a hemifield behavioral paradigm with words and faces as stimuli, with concurrent ERP measurement, in a group of adults with developmental dyslexia (DD) or with congenital prosopagnosia (CP), and matched control participants. Behaviorally, the DD group exhibited clear deficits in both word and face processing relative to controls, while the CP group showed a specific deficit in face processing only. This pattern was mirrored in the ERP data too. The DD group evinced neither the normal ERP pattern of RH dominance for faces nor the LH dominance for words. In contrast, the CP group showed the typical ERP superiority for words in the LH but did not show the typical RH superiority for faces. These findings are consistent with the hypothesis that the typical hemispheric organization for words can develop in the absence of typical hemispheric organization for faces but not vice versa, supporting the account of interactive perceptual development.

Alarm Odor Compounds of the Brown Marmorated Stink Bug Exhibit Antibacterial Activity.

Some insects release scented compounds as a defense against predators that also exhibit antimicrobial activity. Trans-2-octenal and trans-2-decenal are the major alarm aldehydes responsible for the scent of Halyomorpha halys, the brown marmorated stink bug. Previous research has shown these aldehydes are antifungal and produce an antipredatory effect, but have never been tested for antibacterial activity. We hypothesized that these compounds functioned similarly to the analogous multifunctional action of earwig compounds, so we tested whether these aldehydes could inhibit the growth of bacteria. Disk diffusion assays indicated that these aldehydes significantly inhibited the growth of Methicillin-resistant Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, in vitro. Moreover, mealworm beetles (Tenebrio molitor) coated in stink bug aldehydes showed a substantial reduction in bacterial colonization compared to vehicle-treated insects. These results suggest that brown marmorated stinkbug aldehydes are indeed antibacterial agents and serve a multifunctional role for this insect. Therefore, stinkbug aldehydes may have potential for use as chemical antimicrobials.