New System for Digital to Analog Transformation and Reconstruction of 12-Lead ECGs

Introduction

We describe initial validation of a new system for digital to analog conversion (DAC) and reconstruction of 12-lead ECGs. The system utilizes an open and optimized software format with a commensurately optimized DAC hardware configuration to accurately reproduce, from digital files, the original analog electrocardiographic signals of previously instrumented patients. By doing so, the system also ultimately allows for transmission of data collected on one manufacturer''s 12-lead ECG hardware/software into that of any other.

Materials and Methods

To initially validate the system, we compared original and post-DAC re-digitized 12-lead ECG data files (∼5-minutes long) in two types of validation studies in 10 patients. The first type quantitatively compared the total waveform voltage differences between the original and re-digitized data while the second type qualitatively compared the automated electrocardiographic diagnostic statements generated by the original versus re-digitized data.

Results

The grand-averaged difference in root mean squared voltage between the original and re-digitized data was 20.8 µV per channel when re-digitization involved the same manufacturer''s analog to digital converter (ADC) as the original digitization, and 28.4 µV per channel when it involved a different manufacturer''s ADC. Automated diagnostic statements generated by the original versus reconstructed data did not differ when using the diagnostic algorithm from the same manufacturer on whose device the original data were collected, and differed only slightly for just 1 of 10 patients when using a third-party diagnostic algorithm throughout.

Conclusion

Original analog 12-lead ECG signals can be reconstructed from digital data files with accuracy sufficient for clinical use. Such reconstructions can readily enable automated second opinions for difficult-to-interpret 12-lead ECGs, either locally or remotely through the use of dedicated or cloud-based servers.     

An early warning system for aquatic environment state monitoring based on an analysis of mussel valve movements

An early warning system for biological monitoring of aquatic environment is proposed, which is based on an analysis of mussel valve movements. It consists of an analog monitor of an original design (analog recorder and analyzer of opening/closing of valves in a group of mussels) and outside devices (multimeters, computers, and mobile phones). In the monitor, an analysis of the state of valve position sensors, summing/averaging of voltage values, and generation of a warning signal are performed in the analog mode. The proposed system (a) diagnoses the state of valve position sensors, (b) determines the voltage representing the average valve opening in a group of mussels (6–12 individuals), (c) generates a warning signal; and, by means of peripheral devices, (d) registers the average voltage of sensors and (e) transmits a warning signal via a mobile communication net. The system can provide an addition (and, sometimes, an alternative) to complicated digital systems analogous to MosselMonitor®.     

The early phase of sodium channel gating current in the squid giant axon

A fast component of displacement current which accompanies the sodium channel gating current has been recorded from the membrane of the giant axon of the squid Loligo forbesii. This component is characterized by relaxation time constants typically shorter than 25 µs. The charge displaced accounts for about 10% (or 2 nC/cm²) of the total displacement charge attributed to voltage-dependent sodium channels. Using a low noise, wide-band voltage clamp system and specially designed voltage step protocols we could demonstrate that this component: (i) is not a recording artifact; (ii) is kinetically independent from the sodium channel activation and inactivation processes; (iii) can account for a significant fraction of the initial amplitude of recorded displacement current and (iv) has a steady state charge transfer which saturates for membrane potentials above + 20 mV and below – 100 mV This component can be modelled as a single step transition using the Eyring-Boltzmann formalism with a quantal charge of 1 e^– and an asymmetrical energy barrier. Furthermore, if it were associated with the squid sodium channel, our data would suggest one fast transition per channel. A possible role as a sodium channel activation trigger, which would still be consistent with kinetic independence, is discussed. Despite uncertainties about its origin, the property of kinetic independence allows subtraction of this component from the total displacement current to reveal a rising phase in the early time course of the remaining current. This will have to be taken into account when modelling the voltage-dependent sodium channel.     

A simple, PC-dedicated, implanted digital PIM-radiotelemetric system. Part 2: The multichannel system.

The paper, which is a continuation of the previous one, describes a six-channel, PC-dedicated implanted telemetry system, including the schematic drawings, and explanation of all the differences between a single channel and multichannel system approach. The scheme of an additional multichannel analog output, that makes a pure analog recording possible, even without the PC connection, is also presented. Specific methods for both parallel (Centronics) and serial (RS232C) port interfacing are demonstrated and the controlling software principles are discussed. The representative recordings obtained from animal experiments of simultaneous (direct and integrated) multichannel intestinal EMG (MMC) are shown. The device can also be used for the ECG, EEG and uterine smooth muscle electrical activity recording.     

Seven-channel digital telemetry system for monitoring and direct computer capturing of biological data

A seven-channel telemetry system for collection and display of biological data is presented. The system can amplify bioelectrical signals in the range of 2 μV to 200 mV and has a bandwidth of 0.1–80 Hz. After multiplexing, the signals are digitized with a resolution of 8 bits. The data are frequency modulated directly on a VHF transmitter. After receiving the data on a VHF receiver, they are routed directly to the RS232 input connector on the PC. Thereby the advantage of direct communication between the transmitter and the PC can be utilized. Expensive analog equipment is avoided and display of the signals on the PC screen as well as signal analysis can be performed. The system has been tested and was found to be stable and highly reliable.     

CMOS microelectrode array for the monitoring of electrogenic cells

Signal degradation and an array size dictated by the number of available interconnects are the two main limitations inherent to standalone microelectrode arrays (MEAs). A new biochip consisting of an array of microelectrodes with fully-integrated analog and digital circuitry realized in an industrial CMOS process addresses these issues. The device is capable of on-chip signal filtering for improved signal-to-noise ratio (SNR), on-chip analog and digital conversion, and multiplexing, thereby facilitating simultaneous stimulation and recording of electrogenic cell activity. The designed electrode pitch of 250 microm significantly limits the space available for circuitry: a repeated unit of circuitry associated with each electrode comprises a stimulation buffer and a bandpass filter for readout. The bandpass filter has corner frequencies of 100 Hz and 50 kHz, and a gain of 1000. Stimulation voltages are generated from an 8-bit digital signal and converted to an analog signal at a frequency of 120 kHz. Functionality of the read-out circuitry is demonstrated by the measurement of cardiomyocyte activity. The microelectrode is realized in a shifted design for flexibility and biocompatibility. Several microelectrode materials (platinum, platinum black and titanium nitride) have been electrically characterized. An equivalent circuit model, where each parameter represents a macroscopic physical quantity contributing to the interface impedance, has been successfully fitted to experimental results.     

AlGaN/GaN heterostructures for non-invasive cell electrophysiological measurements

Recently, the ability to create bio-semiconductor hybrid devices has gained much interest for cell activity analysis. AlGaN material system has been demonstrated to be a promising cell-based biosensing platform due to a combination of unique properties, such as chemical inertness, optical transparency and low signal to noise ratios. To investigate the potential application of hybrid cell-AlGaN/GaN field effect transistor for cell electrophysiological monitoring, saos-2 human osteoblast-like cells were cultured in high density in non-metallized gate area of a transparent AlGaN/GaN heterostructure field effect transistor. We implemented and characterized the transistor recording of extracellular voltage in the cell-chip junction using the FET chip. The effect of ion channel blocker TEA on transistor signal was explored in order to test the capability of this hybrid chip for in vitro drug screening bioassay. Finally, the effect of cell adhesion on transistor signal was also studied by applying the protein kinase inhibitor H-7.     

Total Charge Movement per Channel : The Relation between Gating Charge Displacement and the Voltage Sensitivity of Activation

One measure of the voltage dependence of ion channel conductance is the amount of gating charge that moves during activation and vice versa. The limiting slope method, introduced by Almers (Almers, W. 1978. Rev. Physiol. Biochem. Pharmacol. 82:96–190), exploits the relationship of charge movement and voltage sensitivity, yielding a lower limit to the range of single channel gating charge displacement. In practice, the technique is plagued by low experimental resolution due to the requirement that the logarithmic voltage sensitivity of activation be measured at very low probabilities of opening. In addition, the linear sequential models to which the original theory was restricted needed to be expanded to accommodate the complexity of mechanisms available for the activation of channels. In this communication, we refine the theory by developing a relationship between the mean activation charge displacement (a measure of the voltage sensitivity of activation) and the gating charge displacement (the integral of gating current). We demonstrate that recording the equilibrium gating charge displacement as an adjunct to the limiting slope technique greatly improves accuracy under conditions where the plots of mean activation charge displacement and gross gating charge displacement versus voltage can be superimposed. We explore this relationship for a wide variety of channel models, which include those having a continuous density of states, nonsequential activation pathways, and subconductance states. We introduce new criteria for the appropriate use of the limiting slope procedure and provide a practical example of the theory applied to low resolution simulation data.     

The effect of tetrodotoxin on the sodium gating current in the squid giant axon.

The effect of tetrodotoxin (TTX) on the sodium gating current in the squid giant axon was examined by recording the current that flowed at the pulse potential at which the ionic current fell to zero, first in the absence and then in the presence of TTX. The addition of 1 microM TTX to the bathing solution had no consistent effect on the size of the initial peak of the gating current, but resulted in small changes in the timecourse of its subsequent relaxation which were mainly caused by a reduction of about one quarter in the component that has a delayed onset and may possibly arise from changes in the state of ionization of groups in the channel wall when the lumen fills with water. Our findings suggest that the binding of TTX at the outer face of the sodium channel does not interfere with the mechanisms of activation and inactivation by the voltage sensors, but has an allosteric effect on the access of internal cations to the inside of the channel.     

Sensory transduction and frequency selectivity in the basal turn of the guinea-pig cochlea.

Receptor potentials recorded from outer hair cells (OHC) and inner hair cells (IHC) in the basal high-frequency turn were compared. The DC component of the IHC receptor potential is maximized to ensure that IHCS can signal a voltage response to high-frequency tones. The OHC DC component is minimized so that OHCS transduce in the most sensitive region of their operating range. The phase and magnitude of OHC receptor potentials were recorded as an indicator of the magnitude and phase of the energy which is fed back to the basilar membrane to provide the basis for the sharp tuning and fine sensitivity of the cochlea to tones. IHC receptor potentials were recorded to assess the net effect of the feedback on the mechanics of the cochlea. It was concluded that OHCS generate feedback which enhances the IHC responses only at the best frequency. At frequencies below CF, IHC DC responses are elicited only when the OHC AC responses begin to saturate.     

Modulation of a gated ion channel admittance in lipid bilayer membranes.

A future class of amperometric biosensors may utilize gated ion channels such as acetylcholine and glutamate receptors as chemical detection components. In this study, bilayer lipid membranes containing voltage-dependent anion channels (VDAC) were used to model an ion-channel-based biosensor which could continuously monitor AC amperometric changes resulting from induced changes in channel conductance. The in-phase and quadrature components of the induced alternating membrane current were monitored as a function of the applied DC offset voltage which was superimposed on the sinusoidal test voltage. The accuracy and sensitivity of the AC-measured VDAC response was dependent on the magnitude of the AC test voltage relative to the DC offset necessary for channel closure. The VDAC channel appears to be a suitable model protein for AC impedance-based biosensor fabrication.     

基于MIMIC数据库的多参数数据D/A回放系统

文中分析了MIMIC多参数智能监护数据库存储文件的识读方法,该数据库公开发表于PhysioNET,还研究及基于STM32和DAC8568的多参数数据D/A回放系统,包括系统硬件工作原理和系统软件工作原理。MIMIC多参数智能监护数据库的数据文件对于变异性较大的生理参数采用连续采样的方式,而对于相对稳定的生理参数采用周期采样的方式,因此,多参数数据各信号的抽样频率不同。系统首先通过VC++编程读取一个多抽样频率数据并在PC机上显示其波形,然后经过D/A硬件设备进行D/A转换后使用示波器输出其波形,实现输出的波形与原始的波形能同时回放。系统可作为模拟信号源,为远程多参数生理监护仪的研制奠定了基础。     

Voltage gated ionic channels in rat cultured astrocytes, reactive astrocytes and an astrocyte-oligodendrocyte progenitor cell

Astrocytes (both type 1 and type 2), cultured from the central nervous system of newborn or 7 day old rats show voltage gated sodium and potassium channels that are activated when the membrane is depolarized to greater than -40 mV. The sodium channels in these cells have an h-infinity curve similar to that of nodal membranes but the activation (peak current-voltage) curves are shifted along the voltage axis by about +30 mV. These sodium currents are blocked only by high concentrations of tetrodotoxin. The voltage activated potassium currents in both types of astrocyte show at least two components; an inactivating component that is suppressed at holding potentials of greater than -40 mV and a persistent, non-inactivating current. Several types of single channel currents were observed in outside-out membrane patches from type 2 astrocytes. One type of potassium channel showed inactivation on depolarization and may contribute to the whole-cell inactivating current. In contrast, oligodendrocytes showed no obvious voltage gated membrane channels. The properties of the type 2 astrocyte-oligodendrocyte progenitor cell were investigated in two ways: 1) by examination of cells just beginning to differentiate along the "electrically silent" oligodendrocyte pathway or 2) by recording from progenitor cells cultured for 24 hours in the presence of cycloheximide to block the appearance of new membrane channels. In both cases, voltage gated inward (sodium) and outward (potassium) currents were noted. The outward current response showed both an inactivating and a non-inactivating component. Similar voltage activated inward and outward membrane currents were noted in reactive astrocytes freshly isolated (3-6 hours) from lesioned areas of adult rat brains.(ABSTRACT TRUNCATED AT 250 WORDS)     

Supercharging: a method for improving patch-clamp performance.

Patch-clamp performance can be improved without altering the normal headstage configuration described by (Hamill, O. P., A. Marty, E. Neher, B. Sakmann, and F. J. Sigworth, 1981, Pfluegers Arch. Eur. J. Physiol., 391:85-100). The "supercharging" method permits resolution of such fast events as calcium and sodium tail currents. Digital computer modeling and analog electronic simulation were used to identify appropriate shapes for the command voltage and the voltage applied to a capacitor tied to the input of the headstage. The voltage command pulse consists of a step with a brief (5-15 microseconds) rectangular spike on its leading edge. Spike amplitude is a function of the membrane capacitance and the access resistance. The spike drives current through the access resistance and speeds charging of the membrane capacitance, making it possible to complete a voltage step within 5-15 microseconds. Clamping speed is independent of the electrode and feedback resistance over a wide range. The second function of the patch clamp amplifier is current measurement, and good time resolution requires suppression of the capacity transient. This can be accomplished by applying an appropriately shaped voltage to the small capacitor tied to the input of the headstage. Series resistance compensation for ionic current transients does not interfere with supercharging. Although the focus of this paper is on whole cell recording, the supercharging concept may prove useful for single channel and bilayer recording techniques.     

A slow component of gating current in crayfish giant axons resembles inactivation charge movement.

Recent experimental evidence from a number of preparations indicates that sodium channel inactivation may be intrinsically voltage sensitive. Intrinsically voltage sensitive inactivation should produce a charge movement. Crayfish giant axons provide a unique opportunity to reexamine the slower components of gating currents (Ig) for a contribution from inactivation (Igh). In reference to other axon preparations, this preparation has relatively rapid inactivation, and steady-state inactivation has a comparatively steep voltage dependence. As predicted by a two-state scheme for voltage-sensitive sodium channel inactivation, Ig in crayfish axons includes a slow component with time constant comparable to the time constant of decay of the sodium current. Allowing for some delay in its onset (60 microseconds), inactivation as described by this slow component of Ig carries roughly the amount of charge predicted by the voltage dependence of inactivation.     

Improved Infrared-Sensing Running Wheel Systems with an Effective Exercise Activity Indicator

This paper describes an infrared-sensing running wheel (ISRW) system for the quantitative measurement of effective exercise activity in rats. The ISRW system provides superior exercise training compared with commercially available traditional animal running platforms. Four infrared (IR) light-emitting diode/detector pairs embedded around the rim of the wheel detect the rat’s real-time position; the acrylic wheel has a diameter of 55 cm and a thickness of 15 cm, that is, it is larger and thicker than traditional exercise wheels, and it is equipped with a rubber track. The acrylic wheel hangs virtually frictionless, and a DC motor with an axially mounted rubber wheel, which has a diameter of 10 cm, drives the acrylic wheel from the outer edge. The system can automatically train rats to run persistently. The proposed system can determine effective exercise activity (EEA), with the IR sensors (which are connected to a conventional PC) recording the rat exercise behavior. A prototype of the system was verified by a hospital research group performing ischemic stroke experiments on rats by considering middle cerebral artery occlusion. The experimental data demonstrated that the proposed system provides greater neuroprotection in an animal stroke model compared with a conventional treadmill and a motorized running wheel for a given exercise intensity. The quantitative exercise effectiveness indicator showed a 92% correlation between an increase in the EEA and a decrease in the infarct volume. This indicator can be used as a noninvasive and objective reference in clinical animal exercise experiments.     

Mutations within the S4-S5 linker alter voltage sensor constraints in hERG K+ channels

Human ether-a-go-go related gene (hERG) channel gating is associated with slow activation, yet the mechanistic basis for this is unclear. Here, we examine the effects of mutation of a unique glycine residue (G546) in the S4-S5 linker on voltage sensor movement and its coupling to pore gating. Substitution of G546 with residues possessing different physicochemical properties shifted activation gating by ∼−50 mV (with the exception of G546C). With the activation shift taken into account, the time constant of activation was also accelerated, suggesting a stabilization of the closed state by ∼1.6-4.3 kcal/mol (the energy equivalent of one to two hydrogen bonds). Predictions of the α-helical content of the S4-S5 linker suggest that the presence of G546 in wild-type hERG provides flexibility to the helix. Deactivation gating was affected differentially by the G546 substitutions. G546V induced a pronounced slow component of closing that was voltage-independent. Fluorescence measurements of voltage sensor movement in G546V revealed a slow component of voltage sensor return that was uncoupled from charge movement, suggesting a direct effect of the mutation on voltage sensor movement. These data suggest that G546 plays a critical role in channel gating and that hERG channel closing involves at least two independently modifiable reconfigurations of the voltage sensor.     

Azumolene inhibits a component of store-operated calcium entry coupled to the skeletal muscle ryanodine receptor

Dantrolene reduces the elevated myoplasmic Ca(2+) generated during malignant hyperthermia, a pharmacogenetic crisis triggered by volatile anesthetics. Although specific binding of dantrolene to the type 1 ryanodine receptor (RyR1), the Ca(2+) release channel of skeletal muscle sarcoplasmic reticulum, has been demonstrated, there is little evidence for direct dantrolene inhibition of RyR1 channel function. Recent studies suggest store-operated Ca(2+) entry (SOCE) contributes to skeletal muscle function, but the effect of dantrolene on this pathway has not been examined. Here we show that azumolene, an equipotent dantrolene analog, inhibits a component of SOCE coupled to activation of RyR1 by caffeine and ryanodine, whereas the SOCE component induced by thapsigargin is not affected. Our data suggest that azumolene distinguishes between two mechanisms of cellular signaling to SOCE in skeletal muscle, one that is coupled to and one independent from RyR1.     

Axon公司膜片钳系统的漏减分析

目的和方法：采用大鼠海马脑片盲法膜片钳的全细胞记录技术,研究美国Axon公司生产的膜片钳系统（Axopatch放大器和pClamp软件）中几种漏减功能的意义和作用机制,重点对定标P／N漏减（Scaled P/N leak subtraction)、膜片钳放大器漏减以及Clampfit处理软件漏减功能的选择与使用进行分析与比较。结果：Clampex采样软件中的定标P／N漏减功能比P／N漏减功能的噪声要小;放大器漏减功能可漏减单一去极化电压幅度所诱发的漏电流,但不能同时对不同电压幅度系列去极化所产生的稳态漏电流进行追踪漏减;Clampfit漏减功能由于其设定只要膜两侧存在电位差就有漏电流产生,因而不适合在记录电压门控性离子通道电流时对稳态漏电流进行漏减。结论：在研究电压门控性离子通道的性质时,可采用P／N漏减功能或定标P／N漏减功能对稳态漏电流进行漏减,而Clampfit漏减功能是不合适的。