Galectins Control mTOR in Response to Endomembrane Damage.

The Ser/Thr protein kinase mTOR controls metabolic pathways, including the catabolic process of autophagy. Autophagy plays additional, catabolism-independent roles in homeostasis of cytoplasmic endomembranes and whole organelles. How signals from endomembrane damage are transmitted to mTOR to orchestrate autophagic responses is not known. Here we show that mTOR is inhibited by lysosomal damage. Lysosomal damage, recognized by galectins, leads to association of galectin-8 (Gal8) with the mTOR apparatus on the lysosome. Gal8 inhibits mTOR activity through its Ragulator-Rag signaling machinery, whereas galectin-9 activates AMPK in response to lysosomal injury. Both systems converge upon downstream effectors including autophagy and defense against Mycobacterium tuberculosis. Thus, a novel galectin-based signal-transduction system, termed here GALTOR, intersects with the known regulators of mTOR on the lysosome and controls them in response to lysosomal damage. VIDEO ABSTRACT.

Feats: A database of semantic features for early produced noun concepts.

Semantic feature production norms have several desirable characteristics that have supported models of representation and processing in adults. However, several key challenges have limited the use of semantic feature norms in studies of early language acquisition. First, existing norms provide uneven and inconsistent coverage of early-acquired concepts that are typically produced and assessed in children under the age of three, which is a time of tremendous growth of early vocabulary skills. Second, it is difficult to assess the degree to which young children may be familiar with normed features derived from these adult-generated datasets. Third, it has been difficult to adopt standard methods to generate semantic network models of early noun learning. Here, we introduce Feats-a tool that was designed to make headway on these challenges by providing a database, the Language Learning and Meaning Acquisition (LLaMA) lab Noun Norms that extends a widely used set of feature norms McRae et al. Behavior Research Methods 37, 547-559, (2005) to include full coverage of noun concepts on a commonly used early vocabulary assessment. Feats includes several tools to facilitate exploration of features comprising early-acquired nouns, assess the developmental appropriateness of individual features using toddler-accessibility norms, and extract semantic network statistics for individual vocabulary profiles. We provide a tutorial overview of Feats. We additionally validate our approach by presenting an analysis of an overlapping set of concepts collected across prior and new data collection methods. Furthermore, using network graph analyses, we show that the extended set of norms provides novel, reliable results given their enhanced coverage.

Calf cutaneous stimulation generates reflexes in the thigh that can be modified by subthreshold noisy vibration.

Imperceptible tactile noise applied to the skin of the feet enhances posture-correcting cutaneous reflexes. This sensory augmentation technique, stochastic resonance (SR), has not been tested in the less-sensitive hairy skin of the leg for its reflex-enhancement ability. The objectives of this study were to determine whether calf skin stimulation produces cutaneous reflexes and whether noise can modify the reflex. In 20 participants, electrotactile pulse trains were applied at the calf while participants performed submaximal isometric knee extension. To test SR, five different levels of vibrotactile noise were applied simultaneously to the test input. Muscle activity from the vastus lateralis (VL) was analyzed 60-110 ms after stimulation. Reflex ratios were calculated by dividing the reflex peak activity by the prestimulation background muscle activity. A significant reflex response was evoked in 16/20 participants (5.41 ± 2.6% of background muscle activity); these responses varied between individuals with eight being facilitatory and eight being inhibitory. In half of the participants, a new reflex appeared at some level of added noise (n = 10). The average reflex ratio of the study population was significantly higher at the "optimal" noise level (8.61 ± 4.5) than at "baseline" (4.70 ± 5.6) (P = 0.002); the optimal level varied across participants. These results suggest that cutaneous reflexes exist at the VL in response to calf skin stimulation and that SR can change cutaneous reflexes at the leg. This study provides an important first step toward SR application in clinical populations with sensory loss such as individuals with lower extremity amputation.NEW & NOTEWORTHY Our work showed that cutaneous reflexes, known to be present in response to foot sole stimulation, can also be evoked by stimulation of hairy leg skin. In addition, we demonstrated that adding tactile noise can enhance this reflex response. These findings demonstrate proof-of-concept for potential future applications where tactile stimulation, applied to the leg of an individual with amputation, can enhance postural-relevant reflexes. Improving postural control may reduce the risk of falls in this high-risk population.

The role of semantic similarity in verb learning events: Vocabulary-related changes across early development.

Verb meaning is challenging for children to learn across varied events. This study examined how the taxonomic semantic similarity of the nouns in novel verb learning events in a progressive alignment learning condition differed from the taxonomic dissimilarity of nouns in a dissimilar learning condition in supporting near (similar) and far (dissimilar) verb generalization to novel objects in an eye-tracking task. A total of 48 children in two age groups (23 girls; younger: 21-24 months, Mage = 22.1 months; older: 27-30 months: Mage = 28.3 months) who differed in taxonomic vocabulary size were tested. There were no group or learning condition differences in near generalization. The younger group demonstrated better far generalization of verbs learned with semantically dissimilar nouns. The older group demonstrated the opposite pattern, with better far generalization of verbs learned with semantically similar nouns in the progressive alignment condition. These patterns were associated with children's in-category vocabulary knowledge more than other vocabulary measures, including verb vocabulary size. Taxonomic vocabulary knowledge differentially affects verb learning and generalization across development.

Soleus responses to Achilles tendon stimuli are suppressed by heel and enhanced by metatarsal cutaneous stimuli during standing.

KEY POINTS: We examined the influence of cutaneous feedback from the heel and metatarsal regions of the foot sole on the soleus stretch reflex pathway during standing. We found that heel electrical stimuli suppressed and metatarsal stimuli enhanced the soleus vibration response. Follow-up experiments indicated that the interaction between foot sole cutaneous feedback and the soleus vibration response was likely not mediated by presynaptic inhibition and was contingent upon a modulation at the ⍺-motoneuron pool level. The spatially organized interaction between cutaneous feedback from the foot sole and the soleus vibration response provides information about how somatosensory information is combined to appropriately respond to perturbations during standing. ABSTRACT: Cutaneous feedback from the foot sole provides balance-relevant information and has the potential to interact with spinal reflex pathways. In this study, we examined how cutaneous feedback from the foot sole (heel and metatarsals) influenced the soleus response to proprioceptive stimuli during standing. We delivered noisy vibration (10-115 Hz) to the right Achilles tendon while we intermittently applied electrical pulse trains (five 1-ms pulses at 200 Hz, every 0.8-1.0 s) to the skin under either the heel or the metatarsals of the ipsilateral foot sole. We analysed time-dependent (referenced to cutaneous stimuli) coherence and cross-correlations between the vibration acceleration and rectified soleus EMG. Vibration-EMG coherence was observed across a bandwidth of ∼10-80 Hz, and coherence was suppressed by heel but enhanced by metatarsal cutaneous stimuli. Cross-correlations showed soleus EMG was correlated with the vibration (∼40 ms lag) and cross-correlations were also suppressed by heel (from 104-155 ms) but enhanced by metatarsal (from 76-128 ms) stimuli. To examine the neural mechanisms mediating this reflex interaction, we conducted two further experiments to probe potential contributions from (1) presynaptic inhibition, and (2) modulations at the ⍺- and γ-motoneuron pools. Results suggest the cutaneous interactions with the stretch reflex pathway required a modulation at the ⍺-motoneuron pool and were likely not mediated by presynaptic inhibition. These findings demonstrate that foot sole cutaneous information functionally tunes the stretch reflex pathway during the control of upright posture and balance.

Dedicated SNAREs and specialized TRIM cargo receptors mediate secretory autophagy.

Autophagy is a process delivering cytoplasmic components to lysosomes for degradation. Autophagy may, however, play a role in unconventional secretion of leaderless cytosolic proteins. How secretory autophagy diverges from degradative autophagy remains unclear. Here we show that in response to lysosomal damage, the prototypical cytosolic secretory autophagy cargo IL-1ß is recognized by specialized secretory autophagy cargo receptor TRIM16 and that this receptor interacts with the R-SNARE Sec22b to recruit cargo to the LC3-II+ sequestration membranes. Cargo secretion is unaffected by downregulation of syntaxin 17, a SNARE promoting autophagosome-lysosome fusion and cargo degradation. Instead, Sec22b in combination with plasma membrane syntaxin 3 and syntaxin 4 as well as SNAP-23 and SNAP-29 completes cargo secretion. Thus, secretory autophagy utilizes a specialized cytosolic cargo receptor and a dedicated SNARE system. Other unconventionally secreted cargo, such as ferritin, is secreted via the same pathway.

Soleus single motor units show stronger coherence with Achilles tendon vibration across a broad bandwidth relative to medial gastrocnemius units while standing.

To probe the frequency characteristics of somatosensory responses in the triceps surae muscles, we previously applied suprathreshold noisy vibration to the Achilles tendon and correlated it with ongoing triceps surae muscle activity (recorded via surface EMG) during standing. Stronger responses to tendon stimuli were observed in soleus (Sol) relative to medial gastrocnemius (MGas) surface EMG; however, it is unknown whether differences in motor unit activity or limitations of surface EMG could have influenced this finding. Here, we inserted indwelling EMG into Sol and MGas to record the activity of single motor units while we applied noisy vibration (10-115 Hz) to the right Achilles tendon of standing participants. We analyzed the relationship between vibration acceleration and the spike activity of active single motor units through estimates of coherence, gain, phase, and cross-covariance. We also applied sinusoidal vibration at frequencies from 10 to 100 Hz (in 5-Hz increments) to examine whether motor units demonstrate nonlinear synchronization or phase locking at higher frequencies. Relative to MGas single motor units, Sol units demonstrated stronger coherence and higher gain with noisy vibration across a bandwidth of 7-68 Hz, and larger peak-to-peak cross-covariance at all four stimulus amplitudes examined. Sol and MGas motor unit activity was modulated over the time course of the sinusoidal stimuli across all frequencies, but their phase-locking behavior was minimal. These findings suggest Sol plays a prominent role in responding to disturbances transmitted through the Achilles tendon across a broad frequency band during standing.NEW & NOTEWORTHY We examined the relationship between Achilles tendon stimuli and spike times of single soleus (Sol) and medial gastrocnemius (MGas) motor units during standing. Relative to MGas, Sol units demonstrated stronger coherence and higher gain with noisy stimuli across a bandwidth of 7-68 Hz. Sol and MGas units demonstrated minimal nonlinear phase locking with sinusoidal stimuli. These findings indicate Sol plays a prominent role in responding to tendon stimuli across a broad frequency band.

Cutaneous afferent innervation of the human foot sole: what can we learn from single-unit recordings?

Cutaneous afferents convey exteroceptive information about the interaction of the body with the environment and proprioceptive information about body position and orientation. Four classes of low-threshold mechanoreceptor afferents innervate the foot sole and transmit feedback that facilitates the conscious and reflexive control of standing balance. Experimental manipulation of cutaneous feedback has been shown to alter the control of gait and standing balance. This has led to a growing interest in the design of intervention strategies that enhance cutaneous feedback and improve postural control. The advent of single-unit microneurography has allowed the firing and receptive field characteristics of foot sole cutaneous afferents to be investigated. In this review, we consolidate the available cutaneous afferent microneurographic recordings from the foot sole and provide an analysis of the firing threshold, and receptive field distribution and density of these cutaneous afferents. This work enhances the understanding of the foot sole as a sensory structure and provides a foundation for the continued development of sensory augmentation insoles and other tactile enhancement interventions.

Ambroxol Induces Autophagy and Potentiates Rifampin Antimycobacterial Activity.

Host-directed therapy in tuberculosis is a potential adjunct to antibiotic chemotherapy directed at Mycobacterium tuberculosis Ambroxol, a lead compound, emerged from a screen for autophagy-inducing drugs. At clinically relevant doses, ambroxol induced autophagy in vitro and in vivo and promoted mycobacterial killing in macrophages. Ambroxol also potentiated rifampin activity in a murine tuberculosis model.

Losing touch: age-related changes in plantar skin sensitivity, lower limb cutaneous reflex strength, and postural stability in older adults.

Age-related changes in the density, morphology, and physiology of plantar cutaneous receptors negatively impact the quality and quantity of balance-relevant information arising from the foot soles. Plantar perceptual sensitivity declines with age and may predict postural instability; however, alteration in lower limb cutaneous reflex strength may also explain greater instability in older adults and has yet to be investigated. We replicated the age-related decline in sensitivity by assessing monofilament and vibrotactile (30 and 250 Hz) detection thresholds near the first metatarsal head bilaterally in healthy young and older adults. We additionally applied continuous 30- and 250-Hz vibration to drive mechanically evoked reflex responses in the tibialis anterior muscle, measured via surface electromyography. To investigate potential relationships between plantar sensitivity, cutaneous reflex strength, and postural stability, we performed posturography in subjects during quiet standing without vision. Anteroposterior and mediolateral postural stability decreased with age, and increases in postural sway amplitude and frequency were significantly correlated with increases in plantar detection thresholds. With 30-Hz vibration, cutaneous reflexes were observed in 95% of young adults but in only 53% of older adults, and reflex gain, coherence, and cumulant density at 30 Hz were lower in older adults. Reflexes were not observed with 250-Hz vibration, suggesting this high-frequency cutaneous input is filtered out by motoneurons innervating tibialis anterior. Our findings have important implications for assessing the risk of balance impairment in older adults.

Tactile orientation perception: an ideal observer analysis of human psychophysical performance in relation to macaque area 3b receptive fields.

The ability to resolve the orientation of edges is crucial to daily tactile and sensorimotor function, yet the means by which edge perception occurs is not well understood. Primate cortical area 3b neurons have diverse receptive field (RF) spatial structures that may participate in edge orientation perception. We evaluated five candidate RF models for macaque area 3b neurons, previously recorded while an oriented bar contacted the monkey's fingertip. We used a Bayesian classifier to assign each neuron a best-fit RF structure. We generated predictions for human performance by implementing an ideal observer that optimally decoded stimulus-evoked spike counts in the model neurons. The ideal observer predicted a saturating reduction in bar orientation discrimination threshold with increasing bar length. We tested 24 humans on an automated, precision-controlled bar orientation discrimination task and observed performance consistent with that predicted. We next queried the ideal observer to discover the RF structure and number of cortical neurons that best matched each participant's performance. Human perception was matched with a median of 24 model neurons firing throughout a 1-s period. The 10 lowest-performing participants were fit with RFs lacking inhibitory sidebands, whereas 12 of the 14 higher-performing participants were fit with RFs containing inhibitory sidebands. Participants whose discrimination improved as bar length increased to 10 mm were fit with longer RFs; those who performed well on the 2-mm bar, with narrower RFs. These results suggest plausible RF features and computational strategies underlying tactile spatial perception and may have implications for perceptual learning.

Gain and phase of perceived virtual rotation evoked by electrical vestibular stimuli.

Galvanic vestibular stimulation (GVS) evokes a perception of rotation; however, very few quantitative data exist on the matter. We performed psychophysical experiments on virtual rotations experienced when binaural bipolar electrical stimulation is applied over the mastoids. We also performed analogous real whole body yaw rotation experiments, allowing us to compare the frequency response of vestibular perception with (real) and without (virtual) natural mechanical stimulation of the semicircular canals. To estimate the gain of vestibular perception, we measured direction discrimination thresholds for virtual and real rotations. Real direction discrimination thresholds decreased at higher frequencies, confirming multiple previous studies. Conversely, virtual direction discrimination thresholds increased at higher frequencies, implying low-pass filtering of the virtual perception process occurring potentially anywhere between afferent transduction and cortical responses. To estimate the phase of vestibular perception, participants manually tracked their perceived position during sinusoidal virtual and real kinetic stimulation. For real rotations, perceived velocity was approximately in phase with actual velocity across all frequencies. Perceived virtual velocity was in phase with the GVS waveform at low frequencies (0.05 and 0.1 Hz). As frequency was increased to 1 Hz, the phase of perceived velocity advanced relative to the GVS waveform. Therefore, at low frequencies GVS is interpreted as an angular velocity signal and at higher frequencies GVS becomes interpreted increasingly as an angular position signal. These estimated gain and phase spectra for vestibular perception are a first step toward generating well-controlled virtual vestibular percepts, an endeavor that may reveal the usefulness of GVS in the areas of clinical assessment, neuroprosthetics, and virtual reality.

Spinal sensory projection neuron responses to spinal cord stimulation are mediated by circuits beyond gate control.

Spinal cord stimulation (SCS) is a therapy used to treat intractable pain with a putative mechanism of action based on the Gate Control Theory. We hypothesized that sensory projection neuron responses to SCS would follow a single stereotyped response curve as a function of SCS frequency, as predicted by the Gate Control circuit. We recorded the responses of antidromically identified sensory projection neurons in the lumbar spinal cord during 1- to 150-Hz SCS in both healthy rats and neuropathic rats following chronic constriction injury (CCI). The relationship between SCS frequency and projection neuron activity predicted by the Gate Control circuit accounted for a subset of neuronal responses to SCS but could not account for the full range of observed responses. Heterogeneous responses were classifiable into three additional groups and were reproduced using computational models of spinal microcircuits representing other interactions between nociceptive and nonnociceptive sensory inputs. Intrathecal administration of bicuculline, a GABAA receptor antagonist, increased spontaneous and evoked activity in projection neurons, enhanced excitatory responses to SCS, and reduced inhibitory responses to SCS, suggesting that GABAA neurotransmission plays a broad role in regulating projection neuron activity. These in vivo and computational results challenge the Gate Control Theory as the only mechanism underlying SCS and refine our understanding of the effects of SCS on spinal sensory neurons within the framework of contemporary understanding of dorsal horn circuitry.

A physical constraint on perceptual learning: tactile spatial acuity improves with training to a limit set by finger size.

In touch as in vision, perceptual acuity improves with training to an extent that differs greatly across people; even individuals with similar initial acuity may undergo markedly different improvement with training. What accounts for this variability in perceptual learning? We hypothesized that a simple physical characteristic, fingertip surface area, might constrain tactile learning, because previous research suggests that larger fingers have more widely spaced mechanoreceptors. To test our hypothesis, we trained 10 human participants intensively on a tactile spatial acuity task. During 4 d, participants completed 1900 training trials (38 50-trial blocks) in which they discriminated the orientation of square-wave gratings pressed onto the stationary index or ring finger, with auditory feedback provided to signal correct and incorrect responses. We progressively increased task difficulty by shifting to thinner groove widths whenever participants achieved ≥90% correct block performance. We took optical scans to measure surface area from the distal interphalangeal crease to the tip of the finger. Participants' acuity improved markedly on the trained finger and to a lesser extent on the untrained finger. Crucially, we found that participants' tactile spatial acuity improved toward a theoretical optimum set by their finger size; participants with worse initial performance relative to their finger size improved more with training, and posttraining performance was better correlated than pretraining performance with finger size. These results strongly support the hypothesis that tactile perceptual learning is limited by finger size. We suspect that analogous physical constraints on perceptual learning will be found in other sensory modalities.

Consistency and variability in multimodal parent-child social interaction: An at-home study using head-mounted eye trackers.

Real-time attention coordination in parent-toddler dyads is often studied in tightly controlled laboratory settings. These studies have demonstrated the importance of joint attention in scaffolding the development of attention and the types of dyadic behaviors that support early language learning. Little is known about how often these behaviors occur in toddlers' everyday lives. We brought wireless head-mounted eye trackers to families' homes to study the moment-to-moment patterns of toddlers' and parents' visual attention and manual activity in daily routines. Our sample consisted of English- and Spanish-speaking families who all reported being middle- or upper middle-class. Toddlers were 2 to 3 years old. Consistent with the findings from previous laboratory studies, we found variability in how frequently toddlers attended to named objects in two everyday activities-Object Play and Mealtime. We then tested whether parent-toddler joint attention in the seconds before a naming utterance increased toddler's attention on the named object. We found that joint attention accompanied by the attended object being held increased the child's attention to the labeled object during naming. We posit that in the rich, noisy world of toddlers' everyday lives, embodied attention plays a critical role in coordinating dyadic behaviors and creating informative naming moments. Our findings highlight the importance of studying toddlers' natural behavior in the real world. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Critical role of growth medium for detecting drug interactions in Gram-negative bacteria that model in vivo responses.

The rise in infections caused by multidrug-resistant (MDR) bacteria has necessitated a variety of clinical approaches, including the use of antibiotic combinations. Here, we tested the hypothesis that drug-drug interactions vary in different media, and determined which in vitro models best predict drug interactions in the lungs. We systematically studied pair-wise antibiotic interactions in three different media, CAMHB, (a rich lab medium standard for antibiotic susceptibility testing), a urine mimetic medium (UMM), and a minimal medium of M9 salts supplemented with glucose and iron (M9Glu) with three Gram-negative ESKAPE pathogens, Acinetobacter baumannii (Ab), Klebsiella pneumoniae (Kp), and Pseudomonas aeruginosa (Pa). There were pronounced differences in responses to antibiotic combinations between the three bacterial species grown in the same medium. However, within species, PaO1 responded to drug combinations similarly when grown in all three different media, whereas Ab17978 and other Ab clinical isolates responded similarly when grown in CAMHB and M9Glu medium. By contrast, drug interactions in Kp43816, and other Kp clinical isolates poorly correlated across different media. To assess whether any of these media were predictive of antibiotic interactions against Kp in the lungs of mice, we tested three antibiotic combination pairs. In vitro measurements in M9Glu, but not rich medium or UMM, predicted in vivo outcomes. This work demonstrates that antibiotic interactions are highly variable across three Gram-negative pathogens and highlights the importance of growth medium by showing a superior correlation between in vitro interactions in a minimal growth medium and in vivo outcomes. IMPORTANCE: Drug-resistant bacterial infections are a growing concern and have only continued to increase during the SARS-CoV-2 pandemic. Though not routinely used for Gram-negative bacteria, drug combinations are sometimes used for serious infections and may become more widely used as the prevalence of extremely drug-resistant organisms increases. To date, reliable methods are not available for identifying beneficial drug combinations for a particular infection. Our study shows variability across strains in how drug interactions are impacted by growth conditions. It also demonstrates that testing drug combinations in tissue-relevant growth conditions for some strains better models what happens during infection and may better inform combination therapy selection.

Age related changes in skin sensitivity assessed with smartphone vibration testing.

The capacity to perceive tactile input at the fingertips, referred to as tactile sensitivity, is known to diminish with age due to regressive changes to mechanoreceptor density and morphology. Sensitivity is measured as perceptual responses to stimuli of varying intensity. Contrary to traditional sensitivity monitoring instruments, smartphones are uniquely suited for remote assessment and have shown to deliver highly calibrated stimuli along a broad spectrum of intensity, which may improve test reliability. The aim of this study was to evaluate a vibration-emitting smartphone application, the Vibratus App, as a mode of estimating tactile sensory thresholds in the aging adult. The peripheral nerve function of 40 neurologically healthy volunteers (ages 18-71) was measured using monofilaments, a 128-Hz tuning fork, the Vibratus App, and nerve conduction studies (NCS). Between group differences were analyzed to determine each measurement's sensitivity to age. Spearman correlation coefficients depicted the associative strength between hand-held measurements and sensory nerve action potential (SNAP) amplitude. Inter-rater reliability of traditional instruments and the software-operated smartphone were assessed by intraclass correlation coefficient (ICC2,k). Measurements taken with Vibratus App were significantly different between age groups (p < 0.001). The inter-rater reliability of monofilament, smartphone vibration, and tuning fork testing was moderate to good (ICC2,k = 0.65, 0.69, and 0.79, respectively). The findings of this study support further investigation of smartphones as sensitivity monitoring devices for at home monitoring of skin sensitivity.

Universal ion chromatography method for anions in active pharmaceutical ingredients enabled by computer-assisted separation modeling.

Ion Chromatography (IC) is one of the most widely used methods for analyzing ionic species in pharmaceutical samples. A universal IC method that can separate a wide range of different analytes is highly desired as it can save a lot of time for method development and validation processes. Herein we report the development of a universal method for anions in active pharmaceutical ingredients (APIs) using computer-assisted chromatography modeling tools. We have screened three different IC columns (Dionex IonPac AS28-Fast 4 µm, AS19 4 µm and AS11-HC 4 µm) to determine the best suitable column for universal IC method development. A universal IC method was then developed using an AS11-HC 4 µm column to separate 31 most common anionic substances in 36 mins. This method was optimized using LC Simulator and a model which precisely predicts the retention behavior of 31 anions was established. This model demonstrated an excellent match between predicted and experimental analyte retention time (R2 =0.999). To validate this universal IC method, we have studied the stability of sulfite and sulfide analytes in ambient conditions. The method was then validated for a subset of 29 anions using water and organic solvent/water binary solvents as diluents for commercial APIs. This universal IC method provides an efficient and simple way to separate and analyze common anions in APIs. In addition, the method development process combined with LC simulator modeling can be effectively used as a starting point during method development for other ions beyond those investigated in this study.

Pan-cortical cellular imaging in freely behaving mice using a miniaturized micro-camera array microscope (mini-MCAM).

Understanding how circuits in the brain simultaneously coordinate their activity to mediate complex ethnologically relevant behaviors requires recording neural activities from distributed populations of neurons in freely behaving animals. Current miniaturized imaging microscopes are typically limited to imaging a relatively small field of view, precluding the measurement of neural activities across multiple brain regions. Here we present a miniaturized micro-camera array microscope (mini-MCAM) that consists of four fluorescence imaging micro-cameras, each capable of capturing neural activity across a 4.5 mm x 2.55 mm field of view (FOV). Cumulatively, the mini-MCAM images over 30 mm 2 area of sparsely expressed GCaMP6s neurons distributed throughout the dorsal cortex, in regions including the primary and secondary motor, somatosensory, visual, retrosplenial, and association cortices across both hemispheres. We demonstrate cortex-wide cellular resolution in vivo Calcium (Ca 2+ ) imaging using the mini-MCAM in both head-fixed and freely behaving mice.

Predicted and observed mortality from vector-borne disease in small songbirds.

Numerous diseases of wildlife have recently emerged due to trade and travel. However, the impact of disease on wild animal populations has been notoriously difficult to detect and demonstrate, due to problems of attribution and the rapid disappearance of bodies after death. Determining the magnitude of avian mortality from West Nile virus (WNV) is emblematic of these challenges. Although correlational analyses may show population declines coincident with the arrival of the virus, strong inference of WNV as a cause of mortality or a population decline requires additional evidence. We show how integrating field data on mosquito feeding patterns, avian abundance, and seroprevalence can be used to predict relative mortality from vector-borne pathogens. We illustrate the method with a case study on WNV in three species of small songbirds, tufted titmouse (Baeolophus bicolor), Carolina wrens (Thryothorus ludovicianus), and northern cardinals (Cardinalis cardinalis). We then determined mortality, infectiousness, and behavioral response of wrens and titmouse following infection with WNV in laboratory experiments and compared them to a previous study on WNV mortality in cardinals. In agreement with predictions, we found titmouse had the highest mortality from WNV infection, with 100% of eleven birds perishing within seven days after infection. Mortality in wrens was significantly lower at 27% (3/11), but still substantial. Viremia profiles indicated that both species were highly infectious for WNV and could play roles in WNV amplification. These findings suggest that WNV may be killing many small-bodied birds, despite the absence of large numbers of dead birds testing positive for WNV. More broadly, they illustrate a framework for predicting relative mortality in hosts from vector-borne disease.