Table of contents

Volume 26

Number 4, August 2005

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EDITORIAL

E01

Two of the oldest areas of medical physics and biomedical engineering are measurements in living systems and quantitative mathematical modeling of these biological entities. The growth of Physiological Measurement over recent years attests to the fact that the former area is still an important one while the field in general does the same for the latter. The number of manuscripts that we receive and ultimately publish has increased each year resulting in an increase in the number of pages we publish each year as well. Less than two years ago we went from a quarterly publication to one that appeared in postboxes bimonthly, and immediate Web publication of papers once they are accepted brings timely information to our subscribers more rapidly than our print editions. Starting in 2006, we will publish Physiological Measurement monthly, a further indication of the growth in this area and our concern regarding timely publication.

As we grow, we recognize that measurement in biologic systems is also becoming a much broader field. Not only do we measure quantities associated with organs and organisms, but today the field is also concerned with measurements of and within cells and subcellular components as well as the molecules themselves that constitute and control these fundamental biologic structures. Often it is difficult to simply make a measurement and understand the data in such a complex system or to comprehend the very measurement itself without applying the tools of the other long-standing, well-established area of biomedical engineering and medical physics: mathematical modeling and simulation. Thus, it makes great sense to include these areas as they apply to physiological measurement within the scope of this journal. For this reason, we have added 'modeling and simulation as they apply to measurements' as a new bullet-point in our list of topics related to physiological measurement covered by this journal. Are we changing to a modeling journal? No, we are just recognizing the importance of modeling in the measurement process. The Institute of Physics and Engineering in Medicine (IPEM) has other publications that do an excellent job of covering the field of physiological modeling. We want to limit our concern to validated models of measurements, instrumentation and measurement systems, and data analysis. Our editorial staff and editorial board look forward to your submissions and comments on this new topic.

TOPICAL REVIEWS

R99

Rotary blood pumps offer a cost-effective way to assist the failing heart. Relative to their pulsatile cousins, they can consist of remarkably few moving parts, with attendant advantages in reliability. These advantages are realized in full only if the entire assist system is kept maximally simple. Control of the pump must therefore be based on a minimum number of measurement devices. This paper reviews the measurements that are made in the wide range of implantable rotary blood pump designs that are in development for ventricular assist. In a number of these, fluid-mechanical variables are estimated indirectly from measurements of motor speed and current or power. The introduction explains the goals of rotary blood pump control by comparison to the innate properties of the natural heart. Then motor and fluid-mechanical variables that may be transduced are discussed. Methods of indirect estimation of pressure drop and flow-rate are dealt with, followed by ways of detecting unusual states such as inflow obstruction. It is found that detection of these alone can be the basis of an adequate control strategy. Some groups have estimated variables pertaining to the heart that is being assisted, and there has also been work on monitoring the ongoing health of the assist system itself. The review concludes with a brief look at the wider measurement context for the intensive-care facility that proposes to use such devices to provide circulatory support.

R119

Single-molecule imaging and manipulation techniques have evolved in the past decade from mere jaw-dropping attractions to essential laboratory tools. By applying single-molecule methods important insights otherwise unavailable have been obtained on various biomolecular systems. Constantly improving single-molecule imaging techniques keep expanding the scale of the explorable spatial detail, thereby providing possible solutions to getting around the debilitating diffraction limit present in physiological-condition structural investigations. In some areas, such as motor protein studies, single-molecule methods have become part of the routine and essential research toolkit. Entire research fields, such as single-molecule force spectroscopy, have been born. In the present review single-molecule visualization and manipulation methods are reviewed with a focus on proteins. Relevant signals and prominent applications are discussed along with experimental examples and recent important results. Finally, the perspectives of the single-molecule field are explored.

PAPERS

337

and

Electrical impedance endotomography (EIE) is a modality of impedance imaging where the electrodes are located on an insulating core placed at the centre of the region of interest. The absence of a physical limit to the medium surrounding the probe enables the use of remote electrodes. The present study compares the features of 2-lead measurements, where the two pairs of electrodes are located on the probe, to 1-lead measurements, where one of the two injection electrodes and one of the two sensing electrodes are located at a distance far away from the probe. The methodology was the characterization of the sensitivity matrix under the influence of electrode pattern, reconstruction radius and mesh construction. Three mesh constructions, three values of the reconstruction radius and five electrode patterns were compared. The study was carried out in 2D using calculated data. Measurement noise was simulated by an addition of 5% Gaussian white noise. The images were reconstructed using the Tikhonov method and L-curve technique. The results show that the reconstruction mesh and the radius of the reconstruction domain have less influence on the conditioning of the sensitivity matrix than the electrode pattern. Both 1-lead and 2-lead configurations enabled the reconstruction of images of relatively similar quality. Additional selection criteria are expected from hardware considerations.

351

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On the basis of double-wavelet analysis, the paper proposes a method to study interactions in the form of frequency and amplitude modulation in nonstationary multimode data series. Special emphasis is given to the problem of quantifying the strength of modulation for a fast signal by a coexisting slower dynamics and to its physiological interpretation. Application of the approach is demonstrated for a number of model systems, including a model that generates chaotic dynamics. The approach is then applied to proximal tubular pressure data from rat nephrons in order to estimate the degree to which the myogenic dynamics of the afferent arteriole is modulated by the slower tubulo-glomerular dynamics. Our analysis reveals a significantly stronger interaction between the two mechanisms in spontaneously hypertensive rats than in normotensive rats.

363

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In this paper, we consider systolic arterial pressure time series from healthy subjects and chronic heart failure patients, undergoing paced respiration, and show that different physiological states and pathological conditions may be characterized in terms of predictability of time series signals from the underlying biological system. We model time series by the regularized least-squares approach and quantify predictability by the leave-one-out error. We find that the entrainment mechanism connected to paced breath, that renders the arterial blood pressure signal more regular and thus more predictable, is less effective in patients, and this effect correlates with the seriousness of the heart failure. Using a Gaussian kernel, so that all orders of nonlinearity are taken into account, the leave-one-out error separates controls from patients (probability less than 10−7), and alive patients from patients for whom cardiac death occurred (probability less than 0.01).

373

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Continuous monitoring of physiologic vital signs is routine in neurocritical care. However, this patient information is usually only recorded intermittently (most often hourly) in the medical record. It is unclear whether this is sufficient to represent the occurrence of secondary brain insults (SBIs) or whether more frequent data collection will provide more comprehensive information for patient care. In 16 patients, physiologic data were acquired concurrently via two methods: per clinical routine, usually hourly, in the medical record (MR) and every minute via a custom data acquisition system (DA). SBIs were defined as a mean arterial pressure <90 mmHg, an intracranial pressure > 20 mmHg or a temperature >37.5 °C. Number of events, cumulative duration of events and area under the curve (AUC) were compared between the two methods and 95% limits of agreement were assessed for various methods of MR data interpolation. For all three parameters, analysis of the DA and MR data frequently differed with regard to number of events, total duration of events and AUC. MR data tended to underestimate the number of total events. 95% limits of agreement were most narrow for trapezoidal interpolation of MR data, but even these limits were fairly broad. Assessment of secondary brain insults is highly dependent on (1) the temporal resolution of the method used to acquire patient data and on (2) the interpolation method if data are acquired intermittently. High frequency data acquisition may be necessary for more precise evaluation of secondary brain injury in neurocritical care.

387

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In this study, we measured the characteristic renal impedance profiles of Wistar rats and simulated the profiles using an electrical model with three series connected Windkessel blocks containing inductance. It is expected that a complete renal impedance profile ought to provide better physical properties information and have more diagnostic power than the pulsatility (PI) and resistive indices (RI) as a result of frequency dependency. A characteristic peak value at the third harmonic on the renal impedance amplitude curve was observed and the phase curve decreased with increasing harmonic numbers. From least mean square fitted parameters, the three blocks were given distinct physical properties and identified as: (1) the renal artery, (2) the small arteries plus the afferent arteriole and (3) the residual kidney (i.e., the efferent arteriole plus the post glomerular capillary structures). These allocations were made according to respective physical properties reported in previous research. These classifications were further confirmed when we compressed the kidney or infused Ang II. Variations in electrical parameters concurred with the likely affected blood vessels reported. This model describes renal impedance characteristics well; and it provides useful hints on the physical properties of the renal vascular system as well as allows for distinctions in possible physiologically affected locations during functional disturbance. It has potential for development as a clinical non-invasive diagnostic tool.

401

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Spirometry and electrical impedance tomography (EIT) data from 26 healthy subjects (14 males, 12 females) were used to develop a model linking contrast variations in EIT difference images to lung volume changes. Eight recordings, each 64 s long, were made for each subject in four postures (standing, sitting, reclining at 45°, supine) and two breathing modes (quiet tidal and deep breathing). Age, gender and five anthropometric variables were recorded. The database was divided into four subsets. The first subset, data from 22 subjects (12 males, 10 females) recorded in deep breathing mode, was used to create the model. Validation was done with the other subsets: data recorded during quiet tidal breathing in the same 22 subjects, and data recorded in both breathing modes for the other four subjects. A quadratic equation in ΔVP (lung volume changes recorded by the spirometer) provided a very good fit to total contrast changes in the EIT images. The model coefficients were found to depend on posture, gender, thoracic circumference and scapular skin fold. To validate the model, the quadratic equation was inverted to estimate lung volume changes from the EIT images. The estimated changes were then compared to the measured volume changes. Validations with each data subset yielded mean standard errors ranging from 9.3% to 12.4%. The proposed model is a first step in enabling inter individual comparisons of EIT images since: (1) it provides a framework for incorporating the effects of anthropometric variables, gender and posture, and (2) it references the images to a physical quantity (volume) verifiable by spirometry.

413

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The cardio-respiratory signal is a fundamental vital sign used for assessment of a patient's status. Additionally, the cardio-respiratory signal provides a great deal of information to healthcare providers wishing to monitor healthy individuals. The air mattress sensor system allows the measurement of the respiration and heart beat movements without the use of a harness or sensor on the subject's body, which eliminates the difficulties these pose for long term measurements. In order to increase the sensitivity, a differential measurement technique between two air cells was used. The concept of a balancing tube between two air cells is suggested in order to increase the robustness against postural changes during the measurements. With this balancing tube, the meaningful frequency range could be selected using a pneumatic method. A mathematical model was constructed and validation experiments were performed for step and sinusoidal input signals. This technique was applied to measurements of respiration and heart beat movements in the supine posture on the bed, which showed potential for applications in sleep analysis, unconstrained healthcare monitoring and neonate monitoring.

423

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Microdialysis monitoring of cerebral metabolism is now performed in several neuro-intensive care units. Conventional microdialysis utilizes CMA70 catheters with 20 kDa molecular weight cut-off membranes enabling the measurement of small molecules such as glucose, lactate, pyruvate and glutamate. The CMA71 100 kDa molecular weight cut-off microdialysis catheter has recently been introduced to allow detection of larger molecules such as cytokines. The objective of this study was to perform in vitro and in vivo testing of the CMA71 microdialysis catheter, comparing its performance with the CMA70. In vitro comparison studies of three of each catheter using reference analyte solutions, demonstrated equivalent recovery for glucose, lactate, pyruvate and glutamate (range 94–97% for CMA70 and 88–103% for CMA71). In vivo comparison involved intracranial placement of paired CMA70 and CMA71 catheters (through the same cranial access device) in six patients with severe traumatic brain injury. Both catheters were perfused with CNS Perfusion Fluid without dextran at 0.3 µl min−1 with hourly sampling and bedside analysis on a CMA600 microdialysis analyser. The two catheters yielded equivalent results for glucose, lactate, pyruvate, glutamate and lactate/pyruvate ratio. CMA71 microdialysis catheters can, therefore, be used for routine clinical monitoring of extracellular substances, as well as for their intended research role of larger molecular weight protein sampling.

429

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Perioperative mortality in coronary artery bypass grafting is usually caused by reduced left ventricular function due to regional myocardial ischemia or infarction. Post-operative graft occlusion is a well-known problem in coronary surgery. A sensitive tool to detect graft occlusion and monitor myocardial function may give the opportunity to revise malfunctioning grafts before departure from the hospital. This paper describes how a new method can detect cardiac ischemia using a 3-axis piezoelectric accelerometer. In three anesthetized pigs, a 3-axis piezoelectric accelerometer was sutured on the lateral free wall of the left ventricle. The left anterior descending (LAD) was occluded for different time periods and the accelerometer data were sampled with a PC. Short-time Fourier transform was calculated based on the accelerometer time series. The results were visualized using a 2D color-coded time–frequency plot. In the area of occlusion, a change to stronger power of higher harmonics was observed. Consequently, a difference value between the instant frequency pattern and a reference frequency pattern showed a rise in absolute value during the occlusion period. The preliminary results indicate that early recognition of regional cardiac ischemia is possible by analyzing accelerometer data acquired from the three animal trials using the prototype 3-axis accelerometer sensor.

441

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Automated non-invasive blood pressure measuring devices based on the oscillometric technique are used widely for self-measurement and are often used in clinics in place of the manual, auscultatory method. Oscillometry was originally developed for monitoring purposes and there are questions over its suitability for making diagnostic measurements. This study measured the differences between automated devices, in the absence of physiological variability. We studied 19 low-cost, automated, non-invasive blood pressure devices, using a repeatable artificial arm simulator, and measured the within-device repeatability and between-device differences. We found that the devices were repeatable (mean within-device difference 1 mmHg), but between-device differences were 4.4 mmHg (systolic pressure) and 3.6 mmHg (diastolic pressure), for normal and high-normal blood pressures. Individual devices are sufficiently repeatable for clinical trend use, but differences between devices are sufficiently large that they may be misinterpreted as clinically significant.

447

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A coordinated study of the dispersal of water between the various body compartments (stomach and gut, blood stream and tissue) and the similar dispersal kinetics of ethanol and its metabolism has been carried out involving two healthy volunteers using flowing afterglow mass spectrometry, FA-MS, and selected ion flow tube mass spectrometry, SIFT-MS. Thus, using these techniques, the variations of HDO and ethanol in breath, measured in successive single exhalations, were followed in real time after the ingestion of measured quantities of D2O and ethanol in proportion to the body weights of the subjects at the dose rates D2O ∼ 0.283 g kg–1, ethanol ∼0.067 g kg–1. During the FA-MS experimental periods (about 2 h), the dispersion of HDO into the body water and finally its equilibration in the total body water is observed from which total body water for each subject was determined. In the SIFT-MS measurements, the dispersion of ethanol into the body water and its loss via metabolism was observed until the physiological (pre-dose) breath level of ethanol for each individual was restored. A simple linear transformation is used to derive the time variations of the blood levels of HDO and ethanol. This has allowed a comparison of the fractions of the ingested ethanol that are metabolized during first-pass metabolism for the two subjects. Thus, in one subject 30% and in the other subject 40% of the ingested alcohol is metabolized in the first 20 min following ingestion. The good time resolution allowed by non-invasive breath analysis ensures that the rates of processes such as ethanol metabolism can be accurately measured. Simultaneous measurements of breath acetaldehyde (largely formed via the ethanol metabolism) and acetone were also performed during the SIFT-MS single breath exhalations.

459

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Fetal magnetocardiography (fMCG) provides fetal cardiac traces useful for the prenatal monitoring of fetal heart function. In this paper, we describe an analytical model (ACWD) for the automatic detection of cardiac waves boundaries that works on fetal signals reconstructed from fMCG by means of independent component analysis. ACWD was validated for 45 healthy and 4 arrhythmic fetuses ranging from 22 to 37 weeks; ACWD outcomes were compared with the estimates of three independent investigators. Descriptive statistics were used to assess correspondence between the outcomes of the automatic and manual approaches. The parametric two-tailed Pearson correlation test (α = 0.01) was employed to quantify, by means of the coefficients of determination, the amount of common variation between the sequences of intervals quantified automatically and manually. ACWD performances on short and long rhythm strips were investigated. ACWD demonstrated to be a robust tool providing dependable estimates of cardiac intervals and their variability during the third gestational trimester also in case of fetal arrhythmias. SNR and stability of fetal traces were the factors limiting ACWD performances. ACWD computation time, which was approximately 1:600 with respect to the manual procedure, was comparable with the time required for fCTI estimation on averaged beats.

477

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Knowledge of normal cerebrovascular volumetric flow rate (VFR) dynamics is of interest for establishing baselines, and for providing input data to cerebrovascular model studies. Retrospectively gated phase contrast magnetic resonance imaging was used to measure time-resolved VFR waveforms from the two internal carotid arteries (ICA) and two vertebral arteries (VA) of 17 young, normal volunteers (16M:1F) at rest in a supine posture. After normalizing each waveform to its respective cycle-averaged VFR, the timing and amplitude of feature points from the individual waveforms were averaged together to produce archetypal ICA and VA waveform shapes. Despite significant inter-individual differences in cycle-averaged VFR within the ICA compared to VA (275 ± 52 versus 91 ± 18 mL min−1), the respective waveform shapes were qualitatively similar overall. The VA waveform shape did, however, exhibit significantly higher amplitudes (e.g., peak:average VFR of 1.78 ± 0.30 versus 1.66 ± 0.16; p < 0.05) and significantly higher variability both between and within subjects. A significant correlation was observed between peak and cycle-averaged VFR, suggesting that the representative waveform shapes presented here—when scaled by an individual's cycle-averaged VFR—may be used to characterize normal ICA and VA flow rate dynamics. This capability may be of particular utility for studies where cerebrovascular flow dynamics are required, but only average flow rates are available.

489

and

In this paper, we have shown a simple procedure to detect anomalies in the lungs region by electrical impedance tomography. The main aim of the present study is to investigate the possibility of anomaly detection by using neural networks. Radial basis function neural networks are used as classifiers to classify the anomaly as belonging to the anterior or posterior region of the left lung or the right lung. The neural networks are trained and tested with the simulated data obtained by solving the mathematical model equation governing current flow through the simulated thoracic region. The equation solution and model simulation are done with FEMLAB. The effect of adding a higher number of neurons to the hidden layer can be clearly seen by the reduction in classification error. The study shows that there is interaction between the size (radius) and conductivity of anomalies and for some combination of these two factors the classification error of neural networks will be very small.

503

and

The effectiveness of cryosurgery, treatment of tumors by freezing, is highly dependent on knowledge of transient freezing extent, and therefore relies heavily on real-time imaging techniques for monitoring. Electrical impedance tomography (EIT) holds much promise for this application. In cryosurgery there is a three order of magnitude change in impedance across the freezing boundary and there is a priori knowledge of the freezing origin. Furthermore, an EIT image of the tissue can be done prior to the cryosurgery. In this study, we have developed an EIT front tracking reconstruction algorithm which takes advantage of these particular attributes of cryosurgery. The method tracks the freezing interface rather than the impedance distribution in the freezing tissue. In addition to drastically reducing the number of parameters needed to define the image, the computational complexity is further reduced by using the more appropriate boundary element method (BEM) for solution to the forward problem. The front-tracking method was found to converge rapidly and accurately to a variety of simulated phantom images.

517

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The aim of this study was to investigate the nonlinear characteristics of heart rate variability (HRV) for three recumbent positions: the supine, left lateral and right lateral decubitus positions. Recently, using a linear analyses method (for time and frequency domains), the effect of the right lateral decubitus position on vagal modulation has been found to increase parasympathetic activity and decrease sympathetic modulation. Little is known about the nonlinear dynamics of HRV for the three recumbent positions. Therefore, we studied the correlation dimension (CD), the largest Lyapunov exponent (LLE), the sample entropy (SampEn), the approximate entropy (ApEn) and the exponent α of the 1/f α power spectrum as nonlinear characteristics of HRV. In response to the right lateral decubitus position, the CD, LLE, SampEn and ApEn increased significantly in both coronary artery disease (CAD) and control groups. In the linear analyses, the normalized high-frequency power (nHF) increased in the right lateral decubitus position. The CD, LLE, ApEn and SampEn correlated positively to the nHF. The α exponent did not correlate to either linear measure or CD, but correlated negatively to LLE, ApEn and SampEn. Among the three recumbent positions, it was found that the right lateral decubitus position can increase the complexity of the human physiological system and the vagal modulation of the cardiac autonomic nervous system the most.

531

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Pulse oximetry is commonly used as an arterial blood oxygen saturation (SaO2) measure. However, its other serial output, the photoplethysmography (PPG) signal, is not as well studied. Raw PPG signals can be used to estimate cardiovascular measures like pulse transit time (PTT) and possibly heart rate (HR). These timing-related measurements are heavily dependent on the minimal variability in phase delay of the PPG signals. Masimo SET® Rad-9™ and Novametrix Oxypleth oximeters were investigated for their PPG phase characteristics on nine healthy adults. To facilitate comparison, PPG signals were acquired from fingers on the same hand in a random fashion. Results showed that mean PTT variations acquired from the Masimo oximeter (37.89 ms) were much greater than the Novametrix (5.66 ms). Documented evidence suggests that 1 ms variation in PTT is equivalent to 1 mmHg change in blood pressure. Moreover, the PTT trend derived from the Masimo oximeter can be mistaken as obstructive sleep apnoeas based on the known criteria. HR comparison was evaluated against estimates attained from an electrocardiogram (ECG). Novametrix differed from ECG by 0.71 ± 0.58% (p < 0.05) while Masimo differed by 4.51 ± 3.66% (p > 0.05). Modern oximeters can be attractive for their improved SaO2 measurement. However, using raw PPG signals obtained directly from these oximeters for timing-related measurements warrants further investigations.

545

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Gait-related back movements require coordination of multiple extremities including the flexible trunk. Ageing and chronic back pain influence these adjustments. These complex coordinations can advantageously be quantified by information theoretically based communication measures such as the gait information flow (GIF). Nine back pain patients (aged 61 ± 10yr) and 12 controls (aged 38 ± 10yr) were investigated during normal walking across a distance of 300 m. The back movements were measured as distances between characteristic points (cervical spine CS, thoracic spine TS, lumbar spine LS) by the sonoSens® Monitor, a system for mobile motion analysis. Gait information flow and regularity indices (RI1: short prediction horizon of 100 ms, RI2: longer prediction horizon of walking period) were assessed as communication characteristics. All indices were non-parametrically tested for group differences. Sensitivity and specificity were assessed by bivariate logistic regression models. We found regularity indices systematically dependent on measurement points, information flow horizon and groups. In the patients RI1 was increased, but RI2 was decreased in comparison to the control group. These results quantitatively characterize the altered complex communication in the patients. We conclude that ageing and/or chronic back pain related dysfunctions of gait can advantageously be monitored by gait information flow characteristics of back movements measured as distances between characteristics points at the back surface.

555

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This paper describes an open loop feedback intelligent system for neonatal intensive care management. The system provides a tool enabling the user to make the final decision to accept or reject the advice given. The system collects 18 parameters from the bedside monitor and ventilator using a Medical Information Bus (MIB) system. Comparison between the system's recommendations and seven clinical users (three doctors and four nurses) actions was made during monitoring of seven neonates with gestation age of 27–31 weeks for 124.13 h (μ = 17.7329, σ = 5.3843 h, range = 10.40–23.85 h). The validation process compared the recommendations triggered by the system with the user feedback (agree, disagree, wait). The system made 191 recommendations in total, 33 of which (17%) were for ventilation and 158 (83%) for oxygenation. The clinician agreed with the system ventilation decisions in 30 occasions (91%) and in 148 occasions for the system oxygenation decisions (94%). The overall percentage of the agreement between the system and the clinician was 93%.

571

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The ergonomic performance of an integrated set of 17 audible alarm sounds, divided into low, medium and high priority classes (Block et al 2000 J. Clin. Monit. Comput.16 541–6), has been undertaken. The sounds were tested for their ease of learning/recall, and how closely their intrinsic perceived urgency matched to a clinical assessment of urgency. The tests were computer-administered and performed on 21 volunteers aged from 18 to 52, in two sessions a few days apart. Session 1 taught the meanings of the alarm sounds and session 2 measured the performance of the sounds. The mean correct identification rate for the sounds was 48.4% (range 10.3–90.0%) with 97.5% of misidentifications within sound priority class. The urgency correlation was statistically significant (r = 0.85, p < 0.001) with all priority classes included but within priority class correlations were not statistically significant. Poor within priority class performances were ascribable to a priori aspects of the design of the sound system.

NOTES

N9

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Retinal neovascularization is a symptom associated with various diseases revealing ocular fundus manifestation. Often, these neovascularizations originate from retinal hypoxia. A concomitant phenomenon of hypoxia is acidosis. To recognise this would permit the identification and treatment of hypoxic fundus areas long before first vascular modifications are seen. Thus, the goal of this investigation was to elucidate whether sodium fluorescein could be used as a retinal pH indicator. Sodium fluorescein solution was diluted in PBS (ratio: 1:150 000). The pH was varied from 6.5 to 8.6 by supplementation of HCl or NaOH, respectively. The fluorescence was excited by a pulsed diode laser (wavelength: 446 nm, pulse width: 100 ps) and detected by time-correlated single photon counting (TCSPC) technique. A least-squares fit of the measured fluorescence decay versus time by an exponential function results in the fluorescence lifetime. Ten measurements were taken at each pH for statistical analysis. The dependence of the fluorescence lifetime on the temperature and the concentration of sodium fluorescein was investigated in the same way. The fluorescence lifetime was found to rise from 3.775 ns to 4.11 ns with increasing pH (6.5 to 8.6). However, the gradient decreases with increasing pH. We found highly significant differences (Student's t-test, P < 0.0005) of the fluorescence lifetimes for pH values with a mean difference of 0.125 at pH < 7.65 whereas the differences were still significant (P ⩽ 0.02) at pH > 7.65 and mean pH differences of 0.2. The fluorescence lifetime was independent of the temperature (22 °C to 37 °C) and the concentration of sodium fluorescein (dilution 1:150 000 to 1:2000). The fluorescence lifetime of sodium fluorescein depends on the pH but not on temperature and concentration. Thus, the discrimination of areas with retinal acidosis should be possible by combination of the TCSPC technique with scanning laser ophthalmoscopy. Further investigations have to clarify whether the accuracy of the measurement at the fundus in vivo is sufficient.

N13

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The dynamic performance of a new fibre optic sensor intended for measuring physiological fluid pressures is assessed in water. The sensor's sensitivity is evaluated at 23°, 35° and 37 °C against a Millar pressure catheter for sinusoidal pressure inputs with frequency ranging from 0.5 to 10 Hz. We found that sensitivity versus frequency is flat to 6 Hz and decreases slightly between 6 and 10 Hz. The sensitivity is slightly lower at 23 °C than at 37 °C. The reproducibility of measurements is excellent (two separate calibration tests in two consecutive days). The output of the fibre optic system used shows a constant time delay (0.13 s) for all frequencies tested. Experiments suggest that, with current sensor design, its immersion in degassed water prior to use ensures a reliable performance.

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