Ex-vivo systems for neuromodulation: A comparison of ex-vivo and in-vivo large animal nerve electrophysiology.

BACKGROUND: Little research exists on extending ex-vivo systems to large animal nerves, and to the best of our knowledge, there has yet to be a study comparing these against in-vivo data. This paper details the first ex-vivo system for large animal peripheral nerves to be compared with in-vivo results. NEW METHOD: Detailed ex-vivo and in-vivo closed-loop neuromodulation experiments were conducted on pig ulnar nerves. Temperatures from 20 °C to 37 °C were evaluated for the ex-vivo system. The data were analysed in the time and velocity domains, and a regression analysis established how evoked compound action potential amplitude and modal conduction velocity (CV) varied with temperature and time after explantation. MAIN RESULTS: Pig ulnar nerves were sustained ex-vivo up to 5 h post-explantation. CV distributions of ex-vivo and in-vivo data were compared, showing closer correspondence at 37 °C. Regression analysis results also demonstrated that modal CV and time since explantation were negatively correlated, whereas modal CV and temperature were positively correlated. COMPARISON WITH EXISTING METHODS: Previous ex-vivo systems were primarily aimed at small animal nerves, and we are not aware of an ex-vivo system to be directly compared with in-vivo data. This new approach provides a route to understand how ex-vivo systems for large animal nerves can be developed and compared with in-vivo data. CONCLUSION: The proposed ex-vivo system results were compared with those seen in-vivo, providing new insights into large animal nerve activity post-explantation. Such a system is crucial for complementing in-vivo experiments, maximising collected experimental data, and accelerating neural interface development.

Denoising and decoding spontaneous vagus nerve recordings with machine learning.

Neural interfaces that electrically stimulate the peripheral nervous system have been shown to successfully improve symptom management for several conditions, such as epilepsy and depression. A crucial part for closing the loop and improving the efficacy of implantable neuromodulation devices is the efficient extraction of meaningful information from nerve recordings, which can have a low Signal-to-Noise ratio (SNR) and non-stationary noise. In recent years, machine learning (ML) models have shown outstanding performance in regression and classification problems, but it is often unclear how to translate and assess these for novel tasks in biomedical engineering. This paper aims to adapt existing ML algorithms to carry out unsupervised denoising of neural recordings instead. This is achieved by applying bandpass filtering and two novel ML algorithms to in-vivo spontaneous, low-SNR vagus nerve recordings. The performance of each approach is compared using the task of extracting respiratory afferent activity and validated using cross-correlation, MSE, and accuracy in terms of extracting the true respiratory rate. A variational autoencoder (VAE) model in particular produces results that show better correlation with respiratory activity compared to bandpass filtering, highlighting that these models have the potential to preserve relevant features in complex neural recordings.

An in-vitro system for closed loop neuromodulation of peripheral nerves.

Current neuromodulation research relies heavily on in-vivo animal experiments for developing novel devices and paradigms, which can be costly, time-consuming, and ethically contentious. As an alternative to this, in-vitro systems are being developed for examining explanted tissue in a controlled environment. However, these systems are typically tailored for cellular studies. Thus, this paper describes the development of an in-vitro system for electrically recording and stimulating large animal nerves. This is demonstrated experimentally using explanted pig ulnar nerves, which show evoked compound action potentials (eCAPs) when stimulated. These eCAPs were examined both in the time and velocity domain at a baseline temperature of 20° C, and at temperatures increasing up to those seen in-vivo (37°C). The results highlight that as the temperature is increased within the in-vitro system, faster conduction velocities (CVs) similar to those present in-vivo can be observed. To our knowledge, this is the first time an in-vitro peripheral nerve system has been validated against in-vivo data, which is crucial for promoting more widespread adoption of such systems for the optimisation of neural interfaces.

Microfluidics Integration into Low-Noise Multi-Electrode Arrays.

Organ-on-Chip technology is commonly used as a tool to replace animal testing in drug development. Cells or tissues are cultured on a microchip to replicate organ-level functions, where measurements of the electrical activity can be taken to understand how the cell populations react to different drugs. Microfluidic structures are integrated in these devices to replicate more closely an in vivo microenvironment. Research has provided proof of principle that more accurate replications of the microenvironment result in better micro-physiological behaviour, which in turn results in a higher predictive power. This work shows a transition from a no-flow (static) multi-electrode array (MEA) to a continuous-flow (dynamic) MEA, assuring a continuous and homogeneous transfer of an electrolyte solution across the measurement chamber. The process through which the microfluidic system was designed, simulated, and fabricated is described, and electrical characterisation of the whole structure under static solution and a continuous flow rate of 80 µL/min was performed. The latter reveals minimal background disturbance, with a background noise below 30 µVpp for all flow rates and areas. This microfluidic MEA, therefore, opens new avenues for more accurate and long-term recordings in Organ-on-Chip systems.

Electrochemical Impedance Spectroscopy as a Tool for Monitoring Cell Differentiation from Floor Plate Progenitors to Midbrain Neurons in Real Time.

Here shows that electrical impedance spectroscopy can be used as a non-invasive and real time tool to probe cell adhesion and differentiation from midbrain floor plate progenitors into midbrain neurons on Au electrodes coated with human laminin. The electrical data and equivalent circuit modeling are consistent with standard microscopy analysis and reveal that within the first 6 hours progenitor cells sediment and attach to the electrode within 40 hours. Between 40 and 120 hours, midbrain progenitor cells differentiate into midbrain neurons, followed by an electrochemically stable maturation phase. The ability to sense and characterize non-invasively and in real time cell differentiation opens up unprecedented avenues for implantable therapies and differentiation strategies.

Similarities between pandemics and cancer in growth and risk models.

Cancer and pandemics are leading causes of death globally, with severe socioeconomic repercussions. To better understand these repercussions, we investigate similarities between pandemics and cancer and describe the limited growth in number of infections or cancer cells, using mathematical models. For a pandemic, the analysis shows that in most cases, the initial fast growth is followed by a slower decay in the recovery phase. The risk of infection increases due to the airborne virus contact crossing a risk-threshold. For cancers caused by carcinogens, the increasing risk with age and absorbed dose of toxins that cross a risk-threshold, may lead to the disease onset. The time scales are different for both causes of death: years for cancer development and days to weeks for contact with airborne viruses. Contamination by viruses is on a time scale of seconds or minutes. The risk-threshold to get ill and the number-threshold in cancer cells or viruses, may explain the large variability in the outcome. The number of infected persons per day is better represented in log-lin plots instead of the conventional lin-lin plots. Differences in therapies are discussed. Our mathematical investigation between cancer and pandemics reveals a multifactorial correlation between both fragilities and brings us one step closer to understand, timely predict and ultimately diminish the socioeconomic hurdle of both cancer and pandemics.

Human Breast Cancer Cells Demonstrate Electrical Excitability.

Breast cancer is one of the most prevalent types of cancers worldwide and yet, its pathophysiology is poorly understood. Single-cell electrophysiological studies have provided evidence that membrane depolarization is implicated in the proliferation and metastasis of breast cancer. However, metastatic breast cancer cells are highly dynamic microscopic systems with complexities beyond a single-cell level. There is an urgent need for electrophysiological studies and technologies capable of decoding the intercellular signaling pathways and networks that control proliferation and metastasis, particularly at a population level. Hence, we present for the first time non-invasive in vitro electrical recordings of strongly metastatic MDA-MB-231 and weakly/non-metastatic MCF-7 breast cancer cell lines. To accomplish this, we fabricated an ultra-low noise sensor that exploits large-area electrodes, of 2 mm2, which maximizes the double-layer capacitance and concomitant detection sensitivity. We show that the current recorded after adherence of the cells is dominated by the opening of voltage-gated sodium channels (VGSCs), confirmed by application of the highly specific inhibitor, tetrodotoxin (TTX). The electrical activity of MDA-MB-231 cells surpasses that of the MCF-7 cells, suggesting a link between the cells' bioelectricity and invasiveness. We also recorded an activity pattern with characteristics similar to that of Random Telegraph Signal (RTS) noise. RTS patterns were less frequent than the asynchronous VGSC signals. The RTS noise power spectral density showed a Lorentzian shape, which revealed the presence of a low-frequency signal across MDA-MB-231 cell populations with propagation speeds of the same order as those reported for intercellular Ca2+ waves. Our recording platform paves the way for real-time investigations of the bioelectricity of cancer cells, their ionic/pharmacological properties and relationship to metastatic potential.

Ultrasensitive System for Electrophysiology of Cancer Cell Populations: A Review.

Bioelectricity is the electrical activity produced by living organisms. Understanding the role of bioelectricity in a disease context is important as it contributes to both disease diagnosis and therapeutic intervention. Electrophysiology tools work well for neuronal cultures; however, they are limited in their ability to detect the electrical activity of non-neuronal cells, wherein the majority of cancers arise. Electronic structures capable of detecting and modulating signaling, in real-time, in electrically quiescent cells are urgently required. One of the limitations to understanding the role of bioelectricity in cancer is the inability to detect low-level signals. In this study, we review our latest advances in devising bidirectional transducers with large electrode areas and concomitant low impedances. The resulting high sensitivity is demonstrated by the extracellular detection of electrical activity in Rat-C6 glioma and prostate cancer (PC-3) cell populations. By using specific inhibitors, we further demonstrated that the large electrical activity in Rat-C6 glioma populations is acidosis driven. For PC-3 cells, the use of a calcium inhibitor together with the slowly varying nature of the signal suggests that Ca2+ channels are involved in the cohort electrogenicity.

Collective electrical oscillations of a diatom population induced by dark stress.

Diatoms are photosynthetic microalgae, a group with a major environmental role on the planet due to the biogeochemical cycling of silica and global fixation of carbon. However, they can evolve into harmful blooms through a resourceful communication mechanism, not yet fully understood. Here, we demonstrate that a population of diatoms under darkness show quasi-periodic electrical oscillations, or intercellular waves. The origin is paracrine signaling, which is a feedback, or survival, mechanism that counteracts changes in the physicochemical environment. The intracellular messenger is related to Ca2+ ions since spatiotemporal changes in their concentration match the characteristics of the intercellular waves. Our conclusion is supported by using a Ca2+ channel inhibitor. The transport of Ca2+ ions through the membrane to the extracellular medium is blocked and the intercellular waves disappear. The translation of microalgae cooperative signaling paves the way for early detection and prevention of harmful blooms and an extensive range of stress-induced alterations in the aquatic ecosystem.

Extracellular electrical recording of pH-triggered bursts in C6 glioma cell populations.

Glioma patients often suffer from epileptic seizures because of the tumor's impact on the brain physiology. Using the rat glioma cell line C6 as a model system, we performed long-term live recordings of the electrical activity of glioma populations in an ultrasensitive detection method. The transducer exploits large-area electrodes that maximize double-layer capacitance, thus increasing the sensitivity. This strategy allowed us to record glioma electrical activity. We show that although glioma cells are nonelectrogenic, they display a remarkable electrical burst activity in time. The low-frequency current noise after cell adhesion is dominated by the flow of Na+ ions through voltage-gated ion channels. However, after an incubation period of many hours, the current noise markedly increased. This electric bursting phenomenon was not associated with apoptosis because the cells were viable and proliferative during the period of increased electric activity. We detected a rapid cell culture medium acidification accompanying this event. By using specific inhibitors, we showed that the electrical bursting activity was prompted by extracellular pH changes, which enhanced Na+ ion flux through the psalmotoxin 1-sensitive acid-sensing ion channels. Our model of pH-triggered bursting was unambiguously supported by deliberate, external acidification of the cell culture medium. This unexpected, acidosis-driven electrical activity is likely to directly perturb, in vivo, the functionality of the healthy neuronal network in the vicinity of the tumor bulk and may contribute to seizures in glioma patients.

Electrochemical noise and impedance of Au electrode/electrolyte interfaces enabling extracellular detection of glioma cell populations.

Microelectrode arrays (MEA) record extracellular local field potentials of cells adhered to the electrodes. A disadvantage is the limited signal-to-noise ratio. The state-of-the-art background noise level is about 10 µVpp. Furthermore, in MEAs low frequency events are filtered out. Here, we quantitatively analyze Au electrode/electrolyte interfaces with impedance spectroscopy and noise measurements. The equivalent circuit is the charge transfer resistance in parallel with a constant phase element that describes the double layer capacitance, in series with a spreading resistance. This equivalent circuit leads to a Maxwell-Wagner relaxation frequency, the value of which is determined as a function of electrode area and molarity of an aqueous KCl electrolyte solution. The electrochemical voltage and current noise is measured as a function of electrode area and frequency and follow unambiguously from the measured impedance. By using large area electrodes the noise floor can be as low as 0.3 µVpp. The resulting high sensitivity is demonstrated by the extracellular detection of C6 glioma cell populations. Their minute electrical activity can be clearly detected at a frequency below about 10 Hz, which shows that the methodology can be used to monitor slow cooperative biological signals in cell populations.

Avaliaçäo clínico-epidemiológica do foneutrismo na região de Säo José do Rio Preto, Säo Paulo / Clinical and epidemiological evaluation of phononeutrism in the region of Sao Jose do Rio Preto, Sao Paulo

Dentre os acidentes envolvendo animais peçonhentos,aqueles causados por aracnídeos tomam relevância médica,quer pela frequência com que ocorrem,quer pelas complicações que deles podem advir. São descritos alguns aspectos clínico-epidemiológicos de 116 acidentes provocados por aranhas do gênero phoneutria na região de São José do Rio Preto, atendidos pelo CCIHB-FAMERP,no período de 1989 a 1998. A dor local foi relatada em 111 casos(95 por cento), sendo a principal manifestação clínica. Dos casos atendidos,60(51,7 por cento)ocorrera em cidades da região de São José do Rio Preto e os meses de maior prevalência do foneutrismo foram março,abril e maio(53 casos). No Tratamento empregado,o bloqueio anestésico foi realizado em 50 casos(43 por cento),enquanto que a soroterapia foi indicada em dois pacientes avaliados. Somente 28 pacientes(24 por cento)trouxeram aranha no momento do atendimento

Aspectos clínico-evolutivos e sociais decorrentes da ingestäo de raticida: um caso ilustrativo / Clinical and social aspects of rat poison ingestion: an illustrative case

Os quadros de intoxicação por raticida, a base de monofluoracetato de sódio (composto 1080), são graves,raros de diagnóstico precoce difícil e habitualmente baseiam-se na história de ingestão do veneno e achados clínicos. assim,relata-se o caso de M.S.s.,masculino,38 anos,procedente de São José do Rio Preto,que deu entrada na emergência do HB-FAMERP em 27/02/98,com história de ingestão intencional de 50ml do raticida "Mão Branca", a cerca de uma hora antes,com quadro de crise convulsiva e opistótono. Feito tratamento suporte e sintomático iniciais,encaminhou-se o paciente para UTI,evoluindo insatisfatoriamente ao final de 36 horas,apresentou parada cardíaca irreversível as manobras de ressuscitação