Label-Free Functional Imaging of Vagus Nerve Stimulation-Evoked Potentials at the Cortical Surface.

Vagus Nerve Stimulation (VNS) was the first FDA-approved stimulation therapy to treat patients with refractory epilepsy and remains widely used. The mechanisms behind the therapeutic effect of VNS remain unknown but are thought to involve afferent-mediated modulation to cortical circuits 1. In this work, we use a coherent holographic imaging system to characterize vagus nerve evoked potentials (VEPs) in the cortex in response to typical VNS stimulation paradigms, which does not require electrode placement nor any genetic, structural, or functional labels. We find that stimulation amplitude strongly modulates VEPs response magnitude (effect size 0.401), while pulse width has a moderate modulatory effect (effect size 0.127) and frequency has almost no modulatory effect (effect size 0.009) on the evoked potential magnitude. We find mild interaction between pulse width and frequency. This non-contact label-free functional imaging technique could serve as a non-invasive rapid feedback tool to quantify VEPs and could increase the efficacy of VNS in patients with refractory epilepsy.

Design and validation of a low-cost photomodulator for in vivo photoactivation of a mGluR5 inhibitor.

Purpose: Severe side effects prevent the utilization of otherwise promising drugs in treatments. These side effects arise when drugs affect untargeted tissues due to poor target specificity. In photopharmacology, light controls the timing and the location of drug delivery, improving treatment specificity and pharmacokinetic control. Photopharmaceuticals have not seen widespread adoption in part because researchers do not always have access to reliable and reproducible light delivery devices at prices which fit within the larger research budget. Method: In this work, we present a customizable photomodulator for use in both wearable and implantable devices. For experimental validation of the photomodulator, we photolyse JF-NP-26 in rats. Results: We successfully drive in vivo photopharmacology with a tethered photomodulator and demonstrate modifications which enable the photomodulator to operate wirelessly. Conclusion: By documenting our photomodulator development, we hope to introduce researchers to a simple solution which significantly lowers the engineering barriers to photopharmacology research. Graphical abstract: Researchers present a photomodulator, a device designed to facilitate in vivo photopharmacology. They demonstrate the in vivo capabilities of the photomodulator by photoreleasing raseglurant, an mGluR5 inhibitor, to treat pain in an acute rat model and follow this study by showing how to reconfigure the photomodulator to work wirelessly and interface with other biomedical devices. Supplementary Information: The online version contains supplementary material available at 10.1007/s13534-023-00334-3.

Neuromuscular implants: Interfacing with skeletal muscle for improved clinical translation of prosthetic limbs.

After an amputation, advanced prosthetic limbs can be used to interface with the nervous system and restore motor function. Despite numerous breakthroughs in the field, many of the recent research advancements have not been widely integrated into clinical practice. This review highlights recent innovations in neuromuscular implants-specifically those that interface with skeletal muscle-which could improve the clinical translation of prosthetic technologies. Skeletal muscle provides a physiologic gateway to harness and amplify signals from the nervous system. Recent surgical advancements in muscle reinnervation surgeries leverage the "bio-amplification" capabilities of muscle, enabling more intuitive control over a greater number of degrees of freedom in prosthetic limbs than previously achieved. We anticipate that state-of-the-art implantable neuromuscular interfaces that integrate well with skeletal muscle and novel surgical interventions will provide a long-term solution for controlling advanced prostheses. Flexible electrodes are expected to play a crucial role in reducing foreign body responses and improving the longevity of the interface. Additionally, innovations in device miniaturization and ongoing exploration of shape memory polymers could simplify surgical procedures for implanting such interfaces. Once implanted, wireless strategies for powering and transferring data from the interface can eliminate bulky external wires, reduce infection risk, and enhance day-to-day usability. By outlining the current limitations of neuromuscular interfaces along with potential future directions, this review aims to guide continued research efforts and future collaborations between engineers and specialists in the field of neuromuscular and musculoskeletal medicine.

Smart Capsule For Targeted Detection Of Inflammation Levels Inside The Gi Tract.

Effective management of Inflammatory Bowel Disease (IBD) is contingent upon frequent monitoring of inflammation levels at targeted locations within the gastrointestinal (GI) tract. This is crucial for assessing disease progression and detecting potential relapses. To address this need, a novel single-use capsule technology has been devised that enables region-specific inflammation measurement, thereby facilitating repeatable monitoring within the GI tract. The capsule integrates a pH-responsive coating for location-specific activation, a chemiluminescent paper-based myeloperoxidase (MPO) sensor for inflammation detection, and a miniaturized photodetector, complemented by embedded electronics for real-time wireless data transmission. Demonstrating linear sensitivity within the physiological MPO concentration range, the sensor is capable of effectively identifying inflammation risk in the GI fluid. Luminescence emitted by the sensor, proportional to MPO concentration, is converted into an electrical signal by the photodetector, generating a quantifiable energy output with a sensitivity of 6.14 µJ/U.ml-1. The capsule was also tested with GI fluids collected from pig models simulating various inflammation states. Despite the physiological complexities, the capsule consistently activated in the intended region and accurately detected MPO levels with less than a 5% variation between readings in GI fluid and a PBS solution. This study heralds a significant step towards minimally invasive, in situ GI inflammation monitoring, potentially revolutionizing personalized IBD management and patient-specific therapeutic strategies.

Smart capsule for monitoring inflammation profile throughout the gastrointestinal tract.

Inflammatory bowel disease (IBD) has become alarmingly prevalent in the last two decades affecting 6.8 million people worldwide with a starkly high relapse rate of 40% within 1 year of remission. Existing visual endoscopy techniques rely on subjective assessment of images that are error-prone and insufficient indicators of early-stage IBD, rendering them unsuitable for frequent and quantitative monitoring of gastrointestinal health necessary for detecting regular relapses in IBD patients. To address these limitations, we have implemented a miniaturized smart capsule (2.2 cm × 11 mm) that allows monitoring reactive oxygen species (ROS) levels as a biomarker of inflammation for quantitative and frequent profiling of inflammatory lesions throughout the gastrointestinal tract. The capsule is composed of a pH and oxidation reduction potential (ORP) sensor to track the capsule's location and ROS levels throughout the gastrointestinal tract, respectively, and an optimized electronic interface for wireless sensing and data communication. The designed sensors provided a linear and stable performance within the physiologically relevant range of the GI tract (pH: 1-8 and ORP: -500 to +500 mV). Additionally, systematic design optimization of the wireless interface electronics offered an efficient sampling rate of 10 ms for long-running measurements up to 48 h for a complete evaluation of the entire gastrointestinal tract. As a proof-of-concept, the capsule the capsule's performance in detecting inflammation risks was validated by conducting tests on in vitro cell culture conditions, simulating healthy and inflamed gut-like environments. The capsule presented here achieves a new milestone in addressing the emerging need for smart ingestible electronics for better diagnosis and treatment of digestive diseases.

Temporal interference current stimulation in peripheral nerves is not driven by envelope extraction.

Background. Electrical neuromodulation remains an effective therapy for multiple neurological disorders. One strategy to electrically stimulate nerves utilizes the interference of multiple high frequency waveforms. This technique, known as temporal interference stimulation or interferential current stimulation, has recently gained significant attention as a method to improve the state-of-the-art in neurostimulation in both animal studies and human clinical trials.Objective.Here we report our investigation into the fundamental properties of the neuronal response to these types of waveforms-the effects of carrier and envelope frequencies, thresholds, firing behavior, and phase and asymmetric interference patterns.Methods.We utilized a cuff electrode on the rat sciatic nerve to apply a variety of interferential signals. We recorded muscle activity in the plantar muscles and biceps femoris, which are proxies for activity on two of the major branches of the sciatic, which are spatially distinct in the target volume. We tested both fundamental recruitment properties as well as spatial techniques to selectively activate either muscle group.Results.Our data suggest, contrary to the currently accepted explanation, that neurons do not extract envelopes at all, and that the response to these signals is well explained by a resistor-capacitor (i.e. integrator) membrane with a fixed firing threshold. Basic interference techniques do not change recruitment far from electrodes. Techniques can produce regions of both phasic activation and tonic activation/conduction block.Conclusions.An integrator model suggests that interference techniques are less capable of minimally invasive stimulation for a subcortical brain target than previously thought. Human clinical trials using these techniques should reevaluate their methods. Interference stimulation allows significant target selectivity in a peripheral cuff electrode with targets near electrodes. These techniques can allow spatially distinct regions of phasic firing, tonic firing, conduction block, and no effect.

Carotid body stimulation as a potential intervention in sudden death in epilepsy.

OBJECTIVE: To investigate carotid body (CB) mechanisms related to sudden death during seizure. Ictal activation of oxygen-conserving reflexes (OCRs) can trigger fatal cardiorespiratory collapse in seizing rats, which presents like human sudden unexpected death in epilepsy (SUDEP). The CB is strongly implicated in OCR pathways; we hypothesize that modulating CB activity will provide insight into these mechanisms of death. METHODS: Long-Evans rats were anesthetized with urethane. Recordings included: electrocorticography, electrocardiography, respiration via nasal thermocouple, and blood pressure (BP). The mammalian diving reflex (MDR) was activated by cold water delivered through a nasal cannula. Reflex and stimulation trials were repeated up to 16 times (4 pre-intervention, 12 post-intervention) or until death. In some animals, one or both carotid bodies were denervated. In some animals, the CB was electrically stimulated, both with and without MDR. Seizures were induced with kainic acid (KA). RESULTS: Animals without seizure and with no CB modulation survived all reflexes. Non-seizing animals with CB denervation survived 7.1 ± 5.4 reflexes before death, and only 1 of 7 survived past the 12-trial threshold. Electrical CB stimulation without seizure and without reflex caused significant tachypnea and hypotension. Electrical CB stimulation with seizure and without reflex required higher amplitudes to replicate the physiological responses seen outside seizure. Seizing animals without CB intervention survived 3.2 ± 3.6 trials (per-reflex survival rate 42.0% ± 44.4%), and 0 of 7 survived past the 12-trial threshold. Seizing animals with electrical CB stimulation survived 10.5 ± 4.7 ictal trials (per-reflex survival rate 86.3% ± 35.0%), and 6 of 8 survived past the 12-trial threshold. SIGNIFICANCE: These results suggest that, during seizure, the ability of the CB to stimulate a restart of respiration is impaired. The CB and its afferents may be relevant to fatal ictal apnea and SUDEP in humans, and CB stimulation may be a relevant intervention technique in these deaths.

Smart soft contact lenses for continuous 24-hour monitoring of intraocular pressure in glaucoma care.

Continuous monitoring of intraocular pressure, particularly during sleep, remains a grand challenge in glaucoma care. Here we introduce a class of smart soft contact lenses, enabling the continuous 24-hour monitoring of intraocular pressure, even during sleep. Uniquely, the smart soft contact lenses are built upon various commercial brands of soft contact lenses without altering their intrinsic properties such as lens power, biocompatibility, softness, transparency, wettability, oxygen transmissibility, and overnight wearability. We show that the smart soft contact lenses can seamlessly fit across different corneal curvatures and thicknesses in human eyes and therefore accurately measure absolute intraocular pressure under ambulatory conditions. We perform a comprehensive set of in vivo evaluations in rabbit, dog, and human eyes from normal to hypertension to confirm the superior measurement accuracy, within-subject repeatability, and user comfort of the smart soft contact lenses beyond current wearable ocular tonometers. We envision that the smart soft contact lenses will be effective in glaucoma care.

An ASIC System for Closed-Loop Blood Pressure Modulation Through Right Cervical Vagus Nerve Stimulation.

OBJECTIVE: Heart disease is the leading cause of death worldwide. Hypertension is an important precursor and the most common risk factor to heart failure. While some patients can control their high blood pressure with pharmaceuticals, many suffer from resistant hypertension, where antihypertensive medications do not achieve the desired outcome. Electrical stimulation is an emerging therapy to modulate blood pressure and integrating it with closed-loop feedback can improve blood pressure control. METHODS: We design and fabricate two application-specific integrated circuits (ASICs) for stimulation and pressure sensing using TSMC's 180 nm MS RF G process. We create a closed-loop system by integrating the ASICs with a microscale pressure sensor and a custom-built Python script and test the full system in six Long Evans rats using vagus nerve stimulation. RESULTS: After calibration and benchtop verification, we prove the functionality of the system in lowering, and maintaining a desired blood pressure in vivo. The system effectively monitors pressure and stimulates when that pressure exceeds the user-determined threshold. CONCLUSION: By combining this stimulation therapy with a pressure sensor, we present a novel closed-loop, electroceutical system that has the potential to monitor and modulate blood pressure. SIGNIFICANCE: We present a drug-free, potentially side-effect-free electroceutical therapeutic for managing resistant hypertension.

A Highly Miniaturized, Chronically Implanted ASIC for Electrical Nerve Stimulation.

We present a wireless, fully implantable device for electrical stimulation of peripheral nerves consisting of a powering coil, a tuning network, a Zener diode, selectable stimulation parameters, and a stimulator IC, all encapsulated in biocompatible silicone. A wireless RF signal at 13.56 MHz powers the implant through the on-chip rectifier. The ASIC, designed in TSMC's 180 nm MS RF G process, occupies an area of less than 1.2 mm2. The IC enables externally selectable current-controlled stimulation through an on-chip read-only memory with a wide range of 32 stimulation parameters (90-750 µA amplitude, 100 µs or 1 ms pulse width, 15 or 50 Hz frequency). The IC generates the constant current waveform using an 8-bit binary weighted DAC and an H-Bridge. At the most power-hungry stimulation parameter, the average power consumption during a stimulus pulse is 2.6 mW with a power transfer efficiency of ∼5.2%. In addition to benchtop and acute testing, we chronically implanted two versions of the device (a design with leads and a leadless design) on two rats' sciatic nerves to verify the long-term efficacy of the IC and the full system. The leadless device had the following dimensions: height of 0.45 cm, major axis of 1.85 cm, and minor axis of 1.34 cm, with similar dimensions for the device with leads. Both devices were implanted and worked for experiments lasting from 21-90 days. To the best of our knowledge, the fabricated IC is the smallest constant-current stimulator that has been tested chronically.

Parylene-C Microbore Tubing: A Simpler Shunt for Reducing Intraocular Pressure.

OBJECTIVE: Current minimally-invasive glaucoma surgery (MIGS) devices promise to control elevated levels of intraocular pressure (IOP) while avoiding many of the downsides of traditional glaucoma surgery. However, there remains room for improvement in performance metrics, including drainage efficacy, device longevity, and time to implant, as outlined by benchmarks set forth by the Audacious Goals Initiative. We introduce a better shunt, which achieves similar or improved pre-clinical safety and efficacy outcomes to commercial MIGS devices, while reducing surgical profile and implantation time. METHODS: We developed a parylene-based microbore glaucoma drainage device capable of modulating IOP via a minimally-invasive implantation procedure. We surgically implanted microbore tubing in five healthy New Zealand White rabbits and measured IOP levels biweekly using handheld applanation tonometry to assess device efficacy in lowering and maintaining IOP. After 6 weeks, the rabbits were euthanized and eyes were enucleated to evaluate inflammatory and histologic response to a foreign-body implant. RESULTS: This device is the only one that fulfills the 10-minute benchmark for implantation time compared to other commercial MIGS devices. In 4 of 5 animals implanted, post-op IOP in the experimental eye dropped by an average of 16.17%. Histopathologic evaluation revealed localized evidence of minor inflammatory reaction and tissue irritation, as well as minimal fibrosis along the tube-tissue interface. CONCLUSION AND SIGNIFICANCE: Based on these findings, this device stands as a promising platform to lowering IOP, particularly in patients with mild to moderate glaucoma requiring no need for cataract intervention, withouteliciting a severe biological response.

Utilizing multimodal imaging to visualize potential mechanism for sudden death in epilepsy.

Sudden death in epilepsy or SUDEP is a fatal condition that accounts for more than 4000 deaths each year. Limited clinical and preclinical data on sudden death suggest critical contributions from autonomic, cardiac, and respiratory pathways. A potential mechanism for such sudden and severe cardiorespiratory dysregulation may be linked to acid reflux-induced laryngospasm. Here, we expand on our previous investigations and utilize a novel multimodal approach to provide visual evidence of acid reflux-initiated cardiorespiratory distress and subsequent sudden death in seizing rats. We used systemic kainic acid to acutely induce seizure activity in Long Evans rats, under urethane anesthesia. We recorded electroencephalography (EEG), electrocardiography (ECG), chest plethysmography, and esophageal pH signals through a multimodal recording platform, during simultaneous fast MRI scans of the rat stomach and esophagus. MRI images, in correlation with electrophysiology data were used to identify seizure progression, stomach acid movement up the esophagus, cardiorespiratory changes, and sudden death. In all cases of sudden death, esophageal pH recordings alongside MRI images visualized stomach acid movement up the esophagus. Severe cardiac (ST segment elevation), respiratory (intermittent apnea) and brain activity (EEG narrowing due to hypoxia) changes were observed only after acid reached larynx, which strongly suggested onset of laryngospasm following acid reflux. The complementary information coming from electrophysiology and fast MRI scans provided insight into the mechanism of esophageal reflux, laryngospasm, obstructive apnea, and subsequent sudden death in seizing animals. The results carry clinical significance as it outlines a potential mechanism that may be relevant to SUDEP in humans.

Clinical Study of the IOPTx™ System - an Electroceutical Wearable to Lower Intraocular Pressure.

Purpose: To investigate the safety and efficacy of the IOPTx™ system - a novel wearable, electroceutical treatment to lower intraocular pressure. Methods: Patients wear the customized contact lens and spectacles of the IOPTx™ system and undergo three 15-minute randomized stimulation trials at different stimulus amplitudes with 15 minutes of rest in between. The parameters for the stimulation trials include a frequency of 50 Hz, a pulse width of 100 µs, and current amplitudes between 90-150 µA. The optometrist measures the intraocular pressure (IOP) before, immediately after, and 15 minutes after the trial, and performs topography, a slit eye examination, and specular microscopy before and after the entire study to check the health of the eye and confirm the safety of the system. Results: The IOPTx™ system successfully modulates a patient's IOP. By testing various currents, we create individual tuning curves examining the effect of the stimulation amplitude on the change in IOP. Each patient may have an optimal dose-response curve and by normalizing to this value, the IOPTx™ system decreased IOP by an average of 17.7% with fifteen minutes of therapy. No Adverse Events or Adverse Device Effects occurred.Conclusions: The results of this clinical case series provide preliminary evidence of efficacy and safety of the IOPTx™ system and its potential usefulness to lower IOP in glaucoma and ocular hypertension.

Ictal activation of oxygen-conserving reflexes as a mechanism for sudden death in epilepsy.

OBJECTIVE: To test the hypothesis that death with physiological parallels to human cases of sudden unexpected death in epilepsy (SUDEP) can be induced in seizing rats by ictal activation of oxygen-conserving reflexes (OCRs). METHODS: Urethane-anesthetized female Long-Evans rats were implanted with electrodes for electrocardiography (ECG), electrocorticography (ECoG), and respiratory thermocouple; venous and arterial cannulas; and a laryngoscope guide and cannula or nasal cannula for activation of the laryngeal chemoreflex (LCR) or mammalian diving reflex (MDR), respectively. Kainic acid injection, either systemic or into the ventral hippocampus, induced prolonged acute seizures. RESULTS: Reflex challenges during seizures caused sudden death in 18 of 20 rats-all MDR rats (10) and all but two LCR rats (8) failed to recover from ictal activation of OCRs and died within minutes of the reflexes. By comparison, 4 of 4 control (ie, nonseizing) rats recovered from 64 induced diving reflexes (16 per rat), and 4 of 4 controls recovered from 64 induced chemoreflexes (16 per rat). Multiple measures were consistent with reports of human SUDEP. Terminal central apnea preceded terminal asystole in all cases. Heart and respiratory rate fluctuations that paralleled those seen in human SUDEP occurred during OCR-induced sudden death, and mean arterial pressure (MAP) was predictive of death, showing a 17 or 15 mm Hg drop (MDR and LCR, respectively) in the 20 s window centered on the time of brain death. OCR activation was never fatal in nonseizing rats. SIGNIFICANCE: These results present a method of inducing sudden death in two seizure models that show pathophysiology consistent with that observed in human cases of SUDEP. This proposed mechanism directly informs previous findings by our group and others in the field; provides a repeatable, inducible animal model for the study of sudden death; and offers a potential explanation for observations made in cases of human SUDEP.

A DSP Architecture for Distortion-Free Evoked Compound Action Potential Recovery in Neural Response Telemetry System.

This paper presents a digital signal processing (DSP) architecture for real-time and distortion-free recovery of electrically-evoked compound action potentials (ECAPs) from stimulus artifacts and periodic noises in bidirectional neural response telemetry (NRT) system. In this DSP architecture, a low computation-cost bidirectional-filtered coherent averaging (BFCA) method is proposed for programmable linear-phase filtering of ECAPs, which can be easily combined with the alternating-polarity (AP) stimulation method to reject stimulus artifacts overlapped with ECAP responses. Design techniques including the configurable folded infinite-impulse response (IIR) filter and division-free averaging are also presented for efficient hardware implementation. Implemented in 180-nm CMOS process, the proposed DSP architecture consumes 10.03-mm2 area and 2.35-mW post-layout simulated power. The efficacy of the DSP architecture in recovering ECAPs from recorded neural data contaminated by overlapped stimulus artifacts and periodic noises is validated in in-vivo electrical nerve stimulations. Experiment results show that compared with the previous coherent averaging technique, the proposed DSP architecture improves the signal-to-noise ratio (SNR) of ECAP responses by 11 dB and achieves a 3.1% waveform distortion that is 17.1× lower.

Mechanisms and prevention of acid reflux induced laryngospasm in seizing rats.

OBJECTIVE: Recent animal work and limited clinical data have suggested that laryngospasm may be involved in the cardiorespiratory collapse seen in sudden unexpected death in epilepsy (SUDEP). In previous work, we demonstrated in an animal model of seizures that laryngospasm and sudden death were always preceded by acid reflux into the esophagus. Here, we expand on that work by testing several techniques to prevent the acid reflux or the subsequent laryngospasm. METHODS: In urethane anesthetized Long Evans rats, we used systemic kainic acid to acutely induce seizure activity. We recorded pH in the esophagus, respiration, electrocorticography activity, and measured the liquid volume in the stomach postmortem. We performed the following three interventions to attempt to prevent acid reflux or laryngospasm and gain insights into mechanisms: fasting animals for 12 h, severing the gastric nerve, and electrical stimulation of either the gastric nerve or the recurrent laryngeal nerve. RESULTS: Seizing animals had significantly more liquid in their stomach. Severing the gastric nerve and fasting animals significantly reduced stomach liquid volume, subsequent acid reflux, and sudden death. Laryngeal nerve stimulation can reverse laryngospasm on demand. Seizing animals are more susceptible to death from stomach acid-induced laryngospasm than nonseizing animals are to artificial acid-induced laryngospasm. SIGNIFICANCE: These results provide insight into the mechanism of acid production and sudden obstructive apnea in this model. These techniques may have clinical relevance if this model is shown to be similar to human SUDEP.

Semi-permanent transcorneal filter support and in vivo surgical implantation technique for open-angle glaucoma treatment.

Primary open-angle glaucoma is a progressive disease affecting nearly 60 million people worldwide which, if left untreated, can lead to optic nerve head damage and complete loss of sight. Current interventions include: pharmaceutical drops, laser surgery, shunts, and bleb; however, these methods provide insufficient long-term efficacy in intraocular pressure management. We developed a semi-permanent, implantable transcorneal duct as a new aid in the treatment of this disease. The duct, composed of an intracorneal stabilizing washer and hollow screw, creates an interface between the anterior chamber and the external environment, allowing for the outflow of excess aqueous humor. We discuss the fluid mechanics behind designing and implementing a filter material capable of preventing the ingress of bacteria and viruses while modifying aqueous humor outflow resistances to pre-glaucomatous levels, finding the effective radius of such a material to be 10.44 µm. After performing surgical implantation in four rabbit eyes, subsequent testing showed successful integration between the screw and washer. Colored saline injections highlighted fluid flow progression out of the eye through the duct, suggesting that the device may be a viable approach to treating high intraocular pressure created by open-angle glaucoma.

Brainstem activity, apnea, and death during seizures induced by intrahippocampal kainic acid in anaesthetized rats.

OBJECTIVE: To investigate how prolonged seizure activity affects cardiorespiratory function and activity of pre-Bötzinger complex, leading to sudden death. METHODS: Urethane-anesthetized female Long-Evans rats were implanted with nasal thermocouple; venous and arterial cannulae; and electrodes for electrocardiography (ECG) and hippocampal, cortical, and brainstem recording. Kainic acid injection into the ventral hippocampus induced status epilepticus. RESULTS: Seizures caused hypertension, tachycardia, and tachypnea punctuated by recurrent transient apneas. Salivation increased considerably: in 11 of 12 rats, liquid with alkaline pH consistent with saliva was expelled from the mouth. Most transient apneas were obstructive: nasal airflow ceased, while, in 83%, efforts to breathe persisted as continued rhythmic activity of respiratory pre-Bötzinger neurons, inspiratory electromyography (EMG), and excursions of the chest wall and abdomen. Blood pressure oscillated in time with respiratory efforts. This pattern also occurred in a minority of cases (16%) of incomplete apnea, but not in rare cases (1%) of transient central apneas. During transient obstructive apneas, the frequency of all inspiratory efforts decreased abruptly by ~30%, suggesting a resetting of the central respiratory rhythm generator. Twenty-two of thirty-one rats died, due either to obstructive apnea (12) or central apnea following progressive slowing of respiration (10). Most rats dying of central apnea had experienced several transient obstructive apneas. Negative DC field potential shifts of the brainstem followed the final breath, consistent with previous reports on spreading depolarization in mouse models. Timing suggests that the DC shift is a consequence rather than cause of respiratory collapse. Cardiac activity continued for tens of seconds. SIGNIFICANCE: Seizure activity in forebrain induces pronounced autonomic activation and disrupts activity in medullary respiratory centers, resulting in death from either obstructive or central apnea. These results directly inform mechanisms of death in status epilepticus, and indirectly provide clues to mechanisms of sudden unexpected death in epilepsy (SUDEP).

A Method of Flexible Micro-Wire Electrode Insertion in Rodent for Chronic Neural Recording and a Device for Electrode Insertion.

Reliable chronic neural recording from focal deep brain structures is impeded by insertion injury and foreign body response, the magnitude of which is correlated with the mechanical mismatch between the electrode and tissue. Thin and flexible electrodes cause less glial scarring and record longer than stiff electrodes. However, the insertion of flexible microelectrodes in brain has been a challenge. Here, a novel insertion method is proposed, and demonstrated, for precise targeting deep brain structures using flexible micro-wire electrodes. The microelectrode is spun and slowly inserted in brain through an appropriate electrode guide. The electrode guide does not penetrate into cortex. Based on two new mechanisms, namely spinning and guided insertion, we have demonstrated successful insertion of 25-micron platinum flexible electrodes about 10-mm deep in rat brains without buckling. We present an electrode insertion device based on the proposed method and demonstrate its use to implant flexible microelectrodes in rat brains. The step-by-step insertion process is described. Microelectrodes were inserted in the Bötzinger complex and respiratory neural activity was recorded acutely in nine rats and chronically in two rats for 50 days.

Characterization of plasma cytokine response to intraperitoneally administered LPS & subdiaphragmatic branch vagus nerve stimulation in rat model.

Vagus nerve stimulation (VNS) has been on the forefront of inflammatory disorder research and has yielded many promising results. Questions remain, however, about the biological mechanisms of such treatments and the inconsistencies in the methods used in research efforts. Here, we aimed to clarify the inflammatory response to intraperitoneal (IP) injections of lipopolysaccharide (LPS) in rats, while analyzing corresponding effects of electrical stimulation to subdiaphragmatic branches (anterior gastric, accessory celiac, and hepatic) of the left vagus nerve. We accomplished an in-depth characterization of the time-varying cytokine cascade response in the serum of 58 rats to an acute IP LPS challenge over a 330-minute period by utilizing curve-fitting and starting point-alignment methods. We then explored the post-LPS neuromodulation effects of electrically stimulating individually cuffed subdiaphragmatic branches. Through our analysis, we found there to be a consistent order of IP LPS cytokine response (IL-10, TNF-α, GM-CSF, IL-17F, IL-6, IL-22, INF-γ). Apart from IL-10, the IP cytokine cascade was more variable in starting time and occurred later than in previously recorded intravenous (IV) challenges. We also found distinct regulatory effects on multiple cytokine levels by each of the three subdiaphragmatic stimulation subsets. While the time-variability of IP LPS use in rats complicates its utility, we have shown it to be a practical, arguably more physiologically relevant method than IV in rats when our methods are used. More importantly, we have shown that selective subdiaphragmatic neurostimulation can be utilized to selectively induce specific effects on inflammation in the body.