Fabrication of chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold for microtia reconstruction.

Fibrin gel-based scaffolds have promising potential for microtia reconstruction. Autologous chondrocytes and chondrocyte cell sheets are frequently used seed cell sources for cartilage tissue engineering. However, the aesthetic outcome of chondrocyte-based microtia reconstruction is still not satisfactory. In this study, we aimed to fabricate the chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold for microtia reconstruction. We designed a unique auricular mold that could fabricate a fibrin gel scaffold resembling human auricle anatomy. Primary chondrocytes were harvested from rabbit auricular cartilage, and chondrocyte cell sheets were developed. Chondrocyte-microtissues were prepared from the cell sheets. The mixture of chondrocytes/chondrocyte-microtissues was laden in fibrin gel during the auricular scaffold fabrication. The protrusions and recessed structure in the auricular scaffold surface were still clearly distinguishable. After a one-week in vitro culture, the 3 D structure and auricular anatomy of the scaffold were retained. And followed by eight-week subcutaneous implantation, cartilaginous tissue was regenerated in the artificial auricular structure as indicated by the results of H&E, Toluidine blue, Safranin O, and type II collagen (immunohistochemistry) staining. Protrusions and depressions of the auricular scaffold were slightly deformed, but the overall auricular anatomy was maintained after 8-week in vivo implantation. Extracellular matrix components content were similar in artificial auricular cartilage and rabbit native auricular cartilage. In conclusion, the mixture of chondrocytes/chondrocyte-microtissues laden fibrin gel auricular scaffold showed a promising potential for cartilaginous tissue regeneration, suggesting this as an effective approach for autologous chondrocyte-based microtia reconstruction.

Special acoustical role of pinna simplifying spatial target localization by the brown long-eared bat Plecotus auritus.

Echolocating bats locate a target by sonar. The performance of this system is related to the shape of the binaural conformation in bats. From numerical predictions, it was found that in a central frequency band, the orientation of a strong sidelobe is shifted nearly linearly in the vertical direction. Inspired by this, the authors built an accurate wide-scope elevation estimation system by constructing a pair of erect artificial pinnae and realized simultaneous elevation and azimuth estimation by constructing a pair of orthogonal pinnae. By demonstrating the simplicity of spatial target echolocation, the authors showed that only two independent single-output neural networks were needed for either elevation or azimuth estimation. This method could be applied to imitate any other mammal species with similar pinna directivity patterns to facilitate and improve an artificial echolocation system.

Spectral cues are necessary to encode azimuthal auditory space in the mouse superior colliculus.

Sound localization plays a critical role in animal survival. Three cues can be used to compute sound direction: interaural timing differences (ITDs), interaural level differences (ILDs) and the direction-dependent spectral filtering by the head and pinnae (spectral cues). Little is known about how spectral cues contribute to the neural encoding of auditory space. Here we report on auditory space encoding in the mouse superior colliculus (SC). We show that the mouse SC contains neurons with spatially-restricted receptive fields (RFs) that form an azimuthal topographic map. We found that frontal RFs require spectral cues and lateral RFs require ILDs. The neurons with frontal RFs have frequency tunings that match the spectral structure of the specific head and pinna filter for sound coming from the front. These results demonstrate that patterned spectral cues in combination with ILDs give rise to the topographic map of azimuthal auditory space.

Variability in the rigid pinna motions of hipposiderid bats and their impact on sensory information encoding.

Many bat species, e.g., in the rhinolophid and hipposiderid families, have dynamic biosonar systems with highly mobile pinnae. Pinna motion patterns have been shown to fall into two distinct categories: rigid rotations and non-rigid motions (i.e., deformations). In the present work, two questions regarding the rigid rotations have been investigated: (i) what is the nature of the variability (e.g., discrete subgroups or continuous variation) within the rigid motions, (ii) what is its acoustic impact? To investigate the first question, rigid pinna motions in Pratt's leaf-nosed bats (Hipposideros pratti) have been tracked with stereo vision and a dense set of landmark points on the pinna surface. Axis-angle representations of the recorded rigid motions have shown a continuous variation in the rotation axes that covered a range of almost 180° in azimuth and elevation. To investigate the second question, the observed range of rigid pinna motions has been reproduced with a biomimetic pinna. Normalized mutual information between acoustic inputs associated with every pair of the rigid pinna motions showed that even small changes in the rotation axis resulted in more than 50% new sensory information encoding capacity (i.e., normalized mutual information less than 50%). This demonstrates a potential sensory benefit to the observed variability in the rigid pinna rotations.

Thermal effects of mobile phones on human auricle region.

Mobile phones have become an indispensable utility to modern society, with international use increasing dramatically each year. The GSM signal operates at 900 MHz, 1800 MHz and 2250 MHz, may potentially cause harm to human tissue. Yet there is no in silico model to aid design these devices to protect from causing potential thermal effect. Here we present a model of sources of heating in a mobile phone device with experimental verification during the phone call. We have developed this mobile phone thermal model using first principles on COMSOL® Multiphysics modelling platform to simulate heating effect in human auricle region due to mobile phone use. In particular, our model considered both radiative and non-radiative heating from components such as the lithium ion battery, CPU circuitry and the antenna. The model showed the distribution and effect of the heating effect due to mobile phone use and considered impact of battery discharge rate, battery capacity, battery cathode material, biological tissue distance, antenna radio-wave frequency and intensity. Furthermore, the lithium ion battery heating was validated during experiments using temperature sensors with an excellent agreement between simulated and experimental data (<1% variation). Mobile phone heating during a typical call has also been simulated and compared with experimental infrared thermographic imaging. Importantly, we found that 1800 MHz frequency of data transmission showed the highest temperature increase in the fat/water phantom used in this simulation. We also successfully compared heating distribution in human auricle region during mobile phone use with clinical thermographic images with reasonable qualitative and quantitative agreements. In summary, our model provides a foundation to conceive thermal and other physical effects caused by mobile phone use and allow for the understanding of potential negative health effects thus supporting and promoting personalized and preventive medicine using thermography.

Gap-induced inhibition of the post-auricular muscle response in humans and guinea pigs.

A common method for measuring changes in temporal processing sensitivity in both humans and animals makes use of GaP-induced Inhibition of the Acoustic Startle (GPIAS). It is also the basis of a common method for detecting tinnitus in rodents. However, the link to tinnitus has not been properly established because GPIAS has not yet been used to objectively demonstrate tinnitus in humans. In guinea pigs, the Preyer (ear flick) myogenic reflex is an established method for measuring the acoustic startle for the GPIAS test, while in humans, it is the eye-blink reflex. Yet, humans have a vestigial remnant of the Preyer reflex, which can be detected by measuring skin surface potentials associated with the Post-Auricular Muscle Response (PAMR). A similar electrical potential can be measured in guinea pigs and we aimed to show that the PAMR could be used to demonstrate GPIAS in both species. In guinea pigs, we compare the GPIAS measured using the pinna movement of the Preyer reflex and the electrical potential of the PAMR to demonstrate that the two are at least equivalent. In humans, we establish for the first time that the PAMR provides a reliable way of measuring GPIAS that is a pure acoustic alternative to the multimodal eye-blink reflex. Further exploratory tests showed that while eye gaze position influenced the size of the PAMR response, it did not change the degree of GPIAS. Our findings confirm that the PAMR is a sensitive method for measuring GPIAS and suggest that it may allow direct comparison of temporal processing between humans and animals and may provide a basis for an objective test of tinnitus.

Nonsurgical treatment of earlobe aging in Mowlavi stages I and II earlobe ptosis with Hyaluronic acid fillers.

BACKGROUND: Earlobe deflation caused by fat atrophy is normally treated with lipofilling, mostly in the context of facelift surgery. In this report, we aim at reporting on Hyaluronic Acid injections to treat earlobe deflation. MATERIALS AND METHODS: 16 Mowlavi Grade I and II patients were treated with HA injections, followed by molding to shape the lobule. RESULTS: Effective correction, lasting 14 months on average, is achieved. Five patients needed a touch-up procedure after 4-6 months to improve the result. CONCLUSIONS: Earlobe augmentation with HA is an ideal option for correction of earlobe atrophy in cases of Mowlavi Grades I and II ptosis. Long-lasting (about 14 months) correction is achieved with no downtime.

Entropy analysis of frequency and shape change in horseshoe bat biosonar.

Echolocating bats use ultrasonic pulses to collect information about their environments. Some of this information is encoded at the baffle structures-noseleaves (emission) and pinnae (reception)-that act as interfaces between the bats' biosonar systems and the external world. The baffle beam patterns encode the direction-dependent sensory information as a function of frequency and hence represent a view of the environment. To generate diverse views of the environment, the bats can vary beam patterns by changes to (1) the wavelengths of the pulses or (2) the baffle geometries. Here we compare the variability in sensory information encoded by just the use of frequency or baffle shape dynamics in horseshoe bats. For this, we use digital and physical prototypes of both noseleaf and pinnae. The beam patterns for all prototypes were either measured or numerically predicted. Entropy was used as a measure to compare variability as a measure of sensory information encoding capacity. It was found that new information was acquired as a result of shape dynamics. Furthermore, the overall variability available for information encoding was similar in the case of frequency or shape dynamics. Thus, shape dynamics allows the horseshoe bats to generate diverse views of the environment in the absence of broadband biosonar signals.

Externalization of remote microphone signals using a structural binaural model of the head and pinna.

In a remote microphone (RM) system, a talker speaks into a microphone and the signal is transmitted to the hearing aids worn by the hearing-impaired listener. A difficulty with remote microphones, however, is that the signal received at the hearing aid bypasses the head and pinna, so the acoustic cues needed to externalize the sound source are missing. The objective of this paper is to process the RM signal to improve externalization when listening through earphones. The processing is based on a structural binaural model, which uses a cascade of processing modules to simulate the interaural level difference, interaural time difference, pinna reflections, ear-canal resonance, and early room reflections. The externalization results for the structural binaural model are compared to a left-right signal blend, the listener's own anechoic head-related impulse response (HRIR), and the listener's own HRIR with room reverberation. The azimuth is varied from straight ahead to 90° to one side. The results show that the structural binaural model is as effective as the listener's own HRIR plus reverberation in producing an externalized acoustic image, and that there is no significant difference in externalization between hearing-impaired and normal-hearing listeners.

Systemic low-frequency oscillations observed in the periphery of healthy human subjects.

This study investigated the relationships of systemic low-frequency oscillations (sLFOs) measured at different peripheral sites in resting state, during passive leg raising (PLR), and during a paced breathing (PB) test. Twenty-five healthy subjects (21 to 57 years old; males: 13 and females: 12) were recruited for these experiments. During the experiments, the fluctuations of oxyhemoglobin concentration were measured at six peripheral sites (left and right toes, fingertips, and earlobes) using a multichannel near-infrared spectroscopy instrument developed by our group. We applied cross-correlation and frequency component analyses on the data. The results showed that the sLFO signals in the symmetric peripheral sites were highly correlated, with time delays close to zero, whereas the correlation coefficients decreased between the sLFO signals of asymmetric sites, with delays up to several seconds. Furthermore, in PLR/PB tests, we found that PB caused wider and more robust changes in hemoglobin concentrations at peripheral sites compared to PLR. Among six peripheral sites, earlobes were the most sensitive to these perturbations, followed by fingertips, and then toes. Lastly, we showed that the perturbation signals may have different coupling mechanisms than the sLFO signals. The study deepened our understanding of the sLFO signals and establishes baseline measures for developing perfusion biomarkers to assess peripheral vascular integrity.

Measuring Pavlovian appetitive conditioning in humans with the postauricular reflex.

Despite its evolutionary and clinical significance, appetitive conditioning has been rarely investigated in humans. It has been proposed that this discrepancy might stem from the difficulty in finding suitable appetitive stimuli that elicit strong physiological responses. However, this might also be due to a possible lack of sensitivity of the psychophysiological measures commonly used to index human appetitive conditioning. Here, we investigated whether the postauricular reflex-a vestigial muscle microreflex that is potentiated by pleasant stimuli relative to neutral and unpleasant stimuli-may provide a valid psychophysiological indicator of appetitive conditioning in humans. To this end, we used a delay differential appetitive conditioning procedure, in which a neutral stimulus was contingently paired with a pleasant odor (CS+), while another neutral stimulus was not associated with any odor (CS-). We measured the postauricular reflex, the startle eyeblink reflex, and skin conductance response (SCR) as learning indices. Taken together, our results indicate that the postauricular reflex was potentiated in response to the CS+ compared with the CS-, whereas this potentiation extinguished when the pleasant odor was no longer delivered. In contrast, we found no evidence for startle eyeblink reflex attenuation in response to the CS+ relative to the CS-, and no effect of appetitive conditioning was observed on SCR. These findings suggest that the postauricular reflex is a sensitive measure of human appetitive conditioning and constitutes a valuable tool for further shedding light on the basic mechanisms underlying emotional learning in humans.

The blastema and epimorphic regeneration in mammals.

Studying regeneration in animals where and when it occurs is inherently interesting and a challenging research topic within developmental biology. Historically, vertebrate regeneration has been investigated in animals that display enhanced regenerative abilities and we have learned much from studying organ regeneration in amphibians and fish. From an applied perspective, while regeneration biologists will undoubtedly continue to study poikilothermic animals (i.e., amphibians and fish), studies focused on homeotherms (i.e., mammals and birds) are also necessary to advance regeneration biology. Emerging mammalian models of epimorphic regeneration are poised to help link regenerative biology and regenerative medicine. The regenerating rodent digit tip, which parallels human fingertip regeneration, and the regeneration of large circular defects through the ear pinna in spiny mice and rabbits, provide tractable, experimental systems where complex tissue structures are regrown through blastema formation and morphogenesis. Using these models as examples, we detail similarities and differences between the mammalian blastema and its classical counterpart to arrive at a broad working definition of a vertebrate regeneration blastema. This comparison leads us to conclude that regenerative failure is not related to the availability of regeneration-competent progenitor cells, but is most likely a function of the cellular response to the microenvironment that forms following traumatic injury. Recent studies demonstrating that targeted modification of this microenvironment can restrict or enhance regenerative capabilities in mammals helps provide a roadmap for eventually pushing the limits of human regeneration.

[Impact of microphone position on sound localization in cochlear implant users]. / Einfluss der Mikrofonposition auf die akustische Lokalisation bei Cochlea-Implantat-Nutzern.

OBJECTIVE: The delivery of directional cues of a hearing device microphone are highly dependent on the position of the microphones. The aim of this work was the evaluation of different microphone positions with regard to the transmission of interaural time and level differences as well as the spectral characteristics and its impact on the localization abilities. METHODS: Head-related transfer functions of 30 subjects were measured with three different omnidirectional microphones at different positions: in the pinna (ITP), behind the ear (BTE), at the entrance of the ear canal (EEC). Sound localization abilities of 12 bilateral CI users was assessed for the microphone positions ITP and BTE. RESULTS: Only the microphone positions in the ear (ITP, EEC) could sample the spectral cues of the pinna. However, the positioning of the microphone inside of the pinna did not significantly improve sound localization abilities compared to BTE microphones. For sound incidence from rear significantly less front-back confusions were achieved with the microphone inside of the pinna. CONCLUSION: The microphone position in the pinna showed only a slight improvement in sound localization compared with BTE microphones in CI users. A precondition for better sound localization abilities is the improvement of the delivery of temporal and spectral fine structure cues in CI systems.

Surgical Denervation of Specific Cutaneous Nerves Impedes Excisional Wound Healing of Small Animal Ear Pinnae.

Damage to cutaneous nerves inhibits wound healing in patients. Results from animals on the nerve contributions to healing are various and sometimes contradictory. Here, we aim to clearly define the collective role of central, caudal, and rostral nerves in ear wound healing of mice, rats, and rabbits. These wounds heal with minimal contraction like wounds in humans. We resected central, caudal, and rostral nerves at the base of ear pinnae by microsurgery and created excisional full-thickness skin wounds in the pinnae neurologically downstream from the resections. Denervation in mice resulted in no closure for 14 days post-wounding (dpw) and led to only 17.2% closure at 21 dpw when the excisional wounds of non-denervated ear pinnae were completely closed. Compared to excisional wounds that were not denervated in sham surgery, wounds with denervation showed an increase of excisional wound areas for 5.0% by 7 dpw and a 43.7% reduction of wound closure at 12 dpw for rats. In rabbits, denervation attenuated wound closure for 14.2, 34.4, and 28.3% at 7, 14, and 18 dpw, respectively. Our histological analysis showed marked denervation impairment in pivotal healing processes, re-epithelialization and granulation tissue growth, suggesting denervation impairment in the regeneration of blood capillaries and/or connective tissue in wounds. These results reveal the critical contributions made by central, caudal, and rostral nerves in ear pinnae to minimal-contraction skin wound healing. Our study also provides small animal models of minimal-contraction wound healing of denervated ear skins that recapitulate human wound healing involving surgical or traumatic nerve damages.

Hind limb motoneurons activity during fictive locomotion or scratching induced by pinna stimulation, serotonin, or glutamic acid in brain cortex-ablated cats.

In brain cortex-ablated cats (BCAC), hind limb motoneurons activity patterns were studied during fictive locomotion (FL) or fictive scratching (FS) induced by pinna stimulation. In order to study motoneurons excitability: heteronymous monosynaptic reflex (HeMR), intracellular recording, and individual Ia afferent fiber antidromic activity (AA) were analyzed. The intraspinal cord microinjections of serotonin or glutamic acid effects were made to study their influence in FL or FS During FS, HeMR amplitude in extensor and bifunctional motoneurons increased prior to or during the respective electroneurogram (ENG). In soleus (SOL) motoneurons were reduced during the scratch cycle (SC). AA in medial gastrocnemius (MG) Ia afferent individual fibers of L6-L7 dorsal roots did not occur during FS Flexor digitorum longus (FDL) and MG motoneurons fired with doublets during the FS bursting activity, motoneuron membrane potential from some posterior biceps (PB) motoneurons exhibits a depolarization in relation to the PB (ENG). It changed to a locomotor drive potential in relation to one of the double ENG, PB bursts. In FDL and semitendinosus (ST) motoneurons, the membrane potential was depolarized during FS, but it did not change during FL Glutamic acid injected in the L3-L4 spinal cord segment favored the transition from FS to FL During FL, glutamic acid produces a duration increase of extensors ENGs. Serotonin increases the ENG amplitude in extensor motoneurons, as well as the duration of scratching episodes. It did not change the SC duration. Segregation and motoneurons excitability could be regulated by the rhythmic generator and the pattern generator of the central pattern generator.

Horseshoe bats and Old World leaf-nosed bats have two discrete types of pinna motions.

Horseshoe bats (Rhinolophidae) and the related Old World leaf-nosed bats (Hipposideridae) both show conspicuous pinna motions as part of their biosonar behaviors. In the current work, the kinematics of these motions in one species from each family (Rhinolophus ferrumequinum and Hipposideros armiger) has been analyzed quantitatively using three-dimensional tracking of landmarks placed on the pinna. The pinna motions that were observed in both species fell into two categories: In "rigid rotations" motions the geometry of the pinna was preserved and only its orientation in space was altered. In "open-close motions" the geometry of the pinna was changed which was evident in a change of the distances between the landmark points. A linear discriminant analysis showed that motions from both categories could be separated without any overlap in the analyzed data set. Hence, bats from both species have two separate types of pinna motions with apparently no transitions between them. The deformations associated with open-close pinna motions in Hipposideros armiger were found to be substantially larger compared to the wavelength associated with the largest pulse energy than in Rhinolophus ferrumequinum (137% vs 99%). The role of the two different motions in the biosonar behaviors of the animals remains to be determined.

Prepulse inhibition and facilitation of the postauricular reflex, a vestigial remnant of pinna startle.

If the postauricular reflex (PAR) is to be used effectively in studies of emotion and attention, its sensitivity to basic modulatory effects such as prepulse inhibition and facilitation must be determined. Two experiments were carried out with healthy young adults to assess the effects of transient and sustained visual prestimuli on the pinna-flexion response to trains of startle probes. In the first experiment, participants passively viewed a small white square. It was displayed from 1,000 ms prior to onset of a train of noise bursts until the end of that train. Relative to no-prepulse control trials, PAR amplitude was inhibited, possibly due to the withdrawal of attentional resources from the auditory modality. In the second experiment, participants performed a visual oddball task in which irrelevant trains of startle probes followed most briefly displayed task stimuli (checkerboards). Prepulse inhibition was observed when a transient stimulus preceded the first probe at a lead time of 100 ms. Amplitude facilitation was observed at longer lead times. In addition to documenting the existence of prepulse inhibition and facilitation, the data suggest that the PAR is not elicited by visual stimuli, that temporal expectancy does not influence its amplitude or latency, and that this vestigial microreflex is resistant to habituation. Results are interpreted in light of a recent theory that the human PAR is a highly degraded pinna startle, in which the reflex arc no longer includes the startle center (nucleus reticularis pontis caudalis).

Chondrogenesis by bone marrow-derived mesenchymal stem cells grown in chondrocyte-conditioned medium for auricular reconstruction.

Bone marrow-derived mesenchymal stem cells (BMSCs) can be obtained by minimally invasive means and would be a favourable source for cell-based cartilage regeneration. However, controlling the differentiation of the BMSCs towards the desired chondrogenic pathway has been a challenge hampering their application. The major aim of the present study was to determine if conditioned medium collected from cultured auricular chondrocytes could promote chondrogenic differentiation of BMSCs. Auricular chondrocytes were isolated and grown in BMSC standard culture medium (SM) that was collected and used as chondrocyte-conditioned medium (CCM). The BMSCs were expanded in either CCM or SM for three passages. Cells were seeded onto fibrous collagen scaffolds and precultured for 2 weeks with or without transforming growth factor-beta 3 (TGF-ß3). After preculture, constructs were implanted subcutaneously in nude mice for 6 and 12 weeks and evaluated with real-time polymerase chain reaction, histology, immunohistochemistry and biochemistry. Real-time polymerase chain reaction results showed upregulation of COL2A1 in the constructs cultured in CCM compared with those in SM. After 12 weeks in vivo, abundant neocartilage formation was observed in the implants that had been cultured in CCM, with or without TGF-ß3. In contrast, very little cartilage matrix formation was observed within the SM groups, regardless of the presence of TGF-ß3. Osteogenesis was only observed in the SM group with TGF-ß3. In conclusion, CCM even had a stronger influence on chondrogenesis than the supplementation of the standard culture medium with TGF-ß3, without signs of endochondral ossification. Efficient chondrogenic differentiation of BMSCs could provide a promising alternative cell population for auricular regeneration. Copyright © 2016 John Wiley & Sons, Ltd.

Vertical normal modes of human ears: Individual variation and frequency estimation from pinna anthropometry.

Beyond the first peak of head-related transfer functions or pinna-related transfer functions (PRTFs) human pinnae are known to have two normal modes with "vertical" resonance patterns, involving two or three pressure anti-nodes in cavum, cymba, and fossa. However, little is known about individual variations in these modes, and there is no established model for estimating their center-frequencies from anthropometry. Here, with geometries of 38 pinnae measured, PRTFs were calculated and vertical modes visualized by numerical simulation. Most pinnae were found to have both Cavum-Fossa and Cavum-Cymba modes, with opposite-phase anti-nodes in cavum and either fossa or cymba, respectively. Nevertheless in both modes, fossa involvement varied substantially across pinnae, dependent on scaphoid fossa depth and cymba shallowness. Linear regression models were evaluated in mode frequency estimation, with 3322 measures derived from 31 pinna landmarks. The Cavum-Fossa normal mode frequency was best estimated [correlation coefficient r = 0.89, mean absolute error (MAE) = 257 Hz or 4.4%] by the distance from canal entrance to helix rim, and cymba horizontal depth. The Cavum-Cymba normal mode frequency was best estimated (r = 0.92, MAE = 247 Hz or 3.2%) by the sagittal-plane distance from concha floor to cymba anterior wall, and cavum horizontal depth.

Postauricular reflexes elicited by soft acoustic clicks and loud noise probes: Reliability, prepulse facilitation, and sensitivity to picture contents.

The startle blink reflex is facilitated during early picture viewing, then inhibited by attention during pleasant and aversive pictures compared to neutral pictures, and finally potentiated during aversive pictures specifically. However, it is unclear whether the postauricular reflex, which is elicited by the same loud acoustic probe as the startle blink reflex but enhanced by appetitive instead of defensive emotion, has the same pattern and time course of emotional modulation. We examined this issue in a sample of 90 undergraduates using serially presented soft acoustic clicks that elicited postauricular (but not startle blink) reflexes in addition to standard startle probes. Postauricular reflexes elicited by both clicks and probes correlated during food and nurturant contents, during which they were potentiated compared to neutral pictures, suggesting clicks effectively elicit emotionally modulated postauricular reflexes. The postauricular reflex was initially facilitated during the first 500 ms of picture processing but was larger during pleasant than neutral pictures throughout picture processing, with larger effect sizes during the latter half of picture processing. Across reflexes and eliciting stimuli, measures of emotional modulation had higher coefficient alphas than magnitudes during specific picture contents within each valence, indicating that only emotional modulation measures assess higher-order appetitive or defensive processing.