Design of a wearable device for physiological parameter monitoring in a COVID setting.

The study focuses on the realization of an accurate device for the detection of different physiological parameters. It has been realized a simple portable system containing the necessary electronics and ensuring the monitoring of the blood oxygenation, the body temperature, the air quality, the respiratory rate and the ECG. The main processing unit consists in a Raspberry Pi Zero W connected to the Healthy Pi4. The latter provides the interface for the clinical pulse-oxymeter while the measures of temperature and quality air are provided using the I2C protocol. The Bluetooth module is finally used to provide the ECG and blood rate data. The collected data are elaborated using Matlab and Python. To evaluate the accuracy of the realized device some experimental tests have been conducted on different subjects, comparing subjects working in Covid area with others resting at home. In both cases the monitoring time was 4 hours. Results have shown good performances of the system, detecting accurately the differences of the parameters values between the two situations. The usability of the device was assessed by administering a questionnaire to the healthcare personnel involved in the experimentation. The outcome shows a good usability of the system as well as an acceptable dressing time.

Real-time optical manipulation of cardiac conduction in intact hearts.

KEY POINTS: Although optogenetics has clearly demonstrated the feasibility of cardiac manipulation, current optical stimulation strategies lack the capability to react acutely to ongoing cardiac wave dynamics. Here, we developed an all-optical platform to monitor and control electrical activity in real-time. The methodology was applied to restore normal electrical activity after atrioventricular block and to manipulate the intraventricular propagation of the electrical wavefront. The closed-loop approach was also applied to simulate a re-entrant circuit across the ventricle. The development of this innovative optical methodology provides the first proof-of-concept that a real-time all-optical stimulation can control cardiac rhythm in normal and abnormal conditions. ABSTRACT: Optogenetics has provided new insights in cardiovascular research, leading to new methods for cardiac pacing, resynchronization therapy and cardioversion. Although these interventions have clearly demonstrated the feasibility of cardiac manipulation, current optical stimulation strategies do not take into account cardiac wave dynamics in real time. Here, we developed an all-optical platform complemented by integrated, newly developed software to monitor and control electrical activity in intact mouse hearts. The system combined a wide-field mesoscope with a digital projector for optogenetic activation. Cardiac functionality could be manipulated either in free-run mode with submillisecond temporal resolution or in a closed-loop fashion: a tailored hardware and software platform allowed real-time intervention capable of reacting within 2 ms. The methodology was applied to restore normal electrical activity after atrioventricular block, by triggering the ventricle in response to optically mapped atrial activity with appropriate timing. Real-time intraventricular manipulation of the propagating electrical wavefront was also demonstrated, opening the prospect for real-time resynchronization therapy and cardiac defibrillation. Furthermore, the closed-loop approach was applied to simulate a re-entrant circuit across the ventricle demonstrating the capability of our system to manipulate heart conduction with high versatility even in arrhythmogenic conditions. The development of this innovative optical methodology provides the first proof-of-concept that a real-time optically based stimulation can control cardiac rhythm in normal and abnormal conditions, promising a new approach for the investigation of the (patho)physiology of the heart.

Effects of ultrasound and selenium on human neurons in vitro.

As the effects of ultrasound on human brain functions might bear therapeutic potential, in this study, we examined the effects of diagnostic, i.e. non-thermal, ultrasound, on morphology, networking, and metabolic activity of SH- SY5Y human neurons in culture, as well as on the expression of GAP-43, Hsp90 and VEGF proteins, with and without selenium in the culture medium. The rationale for studying selenium lays in the observation that selenium improves functional neurologic outcome in traumatic brain injury and, therefore, analysis of the interactions between ultrasound and selenium may be of clinical interest. In the presence of selenium, ultrasound increased the overall number and length of elongations arising from the neuron bodies, thus reflecting an increase in the complexity of neuronal networks and circuits. The expression of GAP-43, Hsp90 and VEGF and metabolic activity of SH-SY5Y neurons, studied as markers of cell damage, were not affected by ultrasound or selenium. This study suggests that ultrasound may modulate neuronal networking in vitro without inducing cellular or molecular damage and highlights the potential role of selenium in the ultrasound-elicited cellular responses.

Resonance artefacts in modern pressure monitoring systems.

Resonance in pressure monitoring catheters is a well-known problem which was studied several years ago. Current piezoelectric devices have mechanical properties providing a resonance frequency and damping factor that theoretically assure resonance-free data. However, in particular cases, the coupling between the device, the catheter, and the vascular compliance of the patient could introduce artefacts in clinical settings leading to wrong pressure waveforms and values displayed in the monitor. In this research work we study a laboratory model of a clinical setting to evaluate in which cases the compound (catheter and device) could cause resonances in an unacceptable range. The classical pop-test is expanded for analysing the effect of the catheter. Results indicate that the presence of different catheters may alter significantly the acquired signal, up to an unacceptable level. Particular care should be used in the selection of the appropriate catheter. In particular, smaller diameters introduce higher damping coefficient that could help in avoiding undesired oscillations.

Shape analysis of the microcirculatory flow wave.

The cardiovascular system and its alterations are a crucial aspect of physiology and medicine. Non-invasive assessment of the functional properties of circulation is of considerable interest to clinicians and physiologists. In this work we investigate the possibility of detecting alterations of the flow waveform in microcirculation, using non-invasive measurements based on a laser Doppler flowmeter. As a test case, we focus on the effect of ageing. Skin is warmed up to a fixed temperature (44 °C) during measurement, to increase blood flow. The shape of the perfusion waveform during each heart beat after the flow was stabilized was used to estimate dynamic parameters of the microcirculatory system. Both the wave rise time, defined as the delay between the diastolic minimum and the following systolic maximum, and the oscillation fraction, defined as the normalized difference between the maximum and minimum flow, present significant variation with age.

Analysis of the microcirculatory pulse wave: age-related alterations.

Morphological analysis of the pulse wave of central blood pressure signal is commonly used for the study of cardiac and vascular properties, but very few attempts were performed for analyzing the peripheral pulse wave of blood flow. In this work, we analyzed this waveform using classical methods, based on the application of FFT, followed by principal components analysis, for assessing the properties of the blood flow. As a sample problem, we evaluated the capability of the proposed method of assessing the alterations correlated with the aging of the vascular system. Results show a good discrimination between the different age groups, confirming the validity of the approach.

Monitoring the microcirculation at the bedside using hand-held imaging microscopes: Automatic tracking of erythrocytes.

In recent years, the development of portable microscopy devices that enable the noninvasive bedside evaluation of the mucosal microcirculation in critically ill patients has expanded the research on this level of the cardiovascular system. Several semi-quantitative scores have been defined to assess the peripheral perfusion, based on videos of the microcirculation. However, they are cumbersome and time-consuming processes, highlighting the need for the development of automated software techniques, to reduce physicians workload and ensure prompt and objective patient evaluation. We developed a rapid, automatic method for the quantitative assessment of erythrocyte velocities in Incident Dark Field (IDF) illumination videos of capillary flow. The technique, based on Kalman filter object tracking, was tested in 35 IDF videos against space-time diagrams, showing high correlation (r = 0.96) and agreement (bias = 3.3 µm/s) with space-time diagrams (STD), below 200 µm/s.

Morphological analysis of neurons: Automatic identification of elongations.

Our study is focused on the development of a new method for the automatic analysis of cell images. We focused on neurons (cells line SH-SY5Y) treated/untreated with ultrasound and stained with Haematoxylin-Eosin. The aim of the algorithm is the automatic detection of the cell body as well as the determination of the number and the length of neuron elongations. Starting point of the algorithm was the convolution of an image with a bank of rotating Gaussian kernels and the construction of a module map. Then several strategies were implemented to detect cell bodies and to detect and extract data about cell elongations. We have also realized a graphical user interface allowing the loading, saving and processing of images. Results show that this method is able to properly and efficiently detect cell contours and elongations. The automated evaluation is in strong agreement with manual evaluation performed by an expert operator, with an average error of 11% with most parameter combinations. This tool constitutes an important support in biological research activities, where operators need to analyze a large number of images to investigate about cell morphology before and after a treatment.

Bioaccumulation of resveratrol metabolites in myocardial tissue is dose-time dependent and related to cardiac hemodynamics in diabetic rats.

BACKGROUND AND AIMS: Trans-resveratrol (RSV) is a natural compound occurring in different foods and plants, which in vivo is rapidly conjugated with glucuronic acid and sulfate. Despite its demonstrated cardioprotective activity, the bioaccumulation of RSV or its metabolites in cardiac tissue is still unknown. METHODS AND RESULTS: Diabetic rats were randomized to 1, 3 or 6 weeks of RSV treatment at two different doses (1 or 5 mg/kg/day). A dose and time-dependent accumulation was observed, with no detectable levels of RSV metabolites found in heart tissues after 1 week and significant concentrations of RSV-3-sulfate and RSV-3-glucuronide after 6 weeks of treatment (0.05 nmol/g of tissue and 0.01 nmol/g of tissue, respectively). Tissue accumulation of RSV metabolites was accompanied by an improvement of cardiac function in long-term diabetes, when myocardial morpho-functional damage is more evident, with an almost complete recovery of all hemodynamic parameters, at the highest RSV dose. CONCLUSION: Even if a higher concentration of RSV in tissues cannot be ruled out after constant oral administration, an accumulation coherent with what is usually evaluated in cell based mechanistic studies is largely unattainable and the RSV unconjugated form would not be present in this paradigm. The current investigation provides data on myocardial tissue concentrations of RSV metabolites, after short/medium term RSV treatment. This knowledge constitutes a basic requirement for future studies aimed at reliably defining the molecular pathways underlying RSV-mediated cardioprotective effects and opens up new perspectives for research focused on testing phenolic compounds as adjuvants in degenerative heart diseases.

DIFFERENCES IN IONIC CURRENTS BETWEEN CANINE MYOCARDIAL AND PURKINJE CELLS.

An electrophysiological analysis of canine single ventricular myocardial (VM) and Purkinje (P) cells was carried out by means of whole cell voltage clamp method. The following results in VM versus P cells were obtained. INa3 was present, had a threshold negative to the fast activating-inactivating INa1, its slow inactivation was cut off by INa1, and contributed to Na+ influx at INa1 threshold. INa1 was smaller and had a less negative threshold. There was no comparable slowly inactivating INa2, accounting for the shorter action potential. Slope conductance at resting potential was about double and decreased to a minimum value at the larger and less negative IK1 peak. The negative slope region of I-V relation was smaller during fast ramps and larger during slow ramps than in P cells, occurred in the voltage range of IK1 block by Mg2+, was not affected by a lower Vh and TTX and was eliminated by Ba2+, in contrast to P cells. ICa was larger, peaked at positive potentials and was eliminated by Ni2+. Ito was much smaller, began at more positive values, was abolished by less negative Vh and by 4-aminopyridine, included a sustained current that 4-aminopyridine decreased but did not eliminate. Steeper ramps increased IK1 peak as well as the fall in outward current during repolarization, consistent with a time-dependent block and unblock of IK1 by polyamines. During repolarization, the positive slope region was consistently present and was similar in amplitude to IK1 peak, whereas it was small or altogether missing in P cells. The total outward current at positive potentials comprised a larger IK1 component whereas it included a larger Ito and sustained current in P cells. These and other results provide a better understanding of the mechanisms underlying the action potential of VM and P cells under normal and some abnormal (arrhythmias) conditions.

Effect of local blood flow in thermal regulation in diabetic patient.

The presence of dysautonomia in diabetic neuropathy is correlated with impairment of vasomotor activity that drives blood microcirculation. Microcirculation, in turn, plays an important role in thermoregulation. In this work, we investigate the changes between two different physiological conditions of diabetic patients, induced by FREMS application, in the control of skin temperature, using a minimally invasive experiment. Skin is warmed up to a fixed temperature (44 °C) for a few minutes, then the heat source is turned off, letting the skin recover its physiological temperature. Both temperature and local blood flow, the latter measured with laser Doppler, are monitored during the experiment. A simple model of the cooling phase is used to evaluate the time constants involved in the process. Results indicate that significant differences exist in the model parameters between the two conditions.

Automatic newborn cry analysis: a non-invasive tool to help autism early diagnosis.

Autism Spectrum Disorders (ASD) are complex developmental disabilities that cause problems with social interaction and communication. ASD are associated with motor development problems, more or less blurred, and with perceptual and sensory brain areas. Crying is the infant's earliest form of communication and recent studies connect this original "language" form with autism disturbs. Being completely non-invasive, cry analysis is an appealing approach for early ASD diagnosis to improve rehabilitation. To this aim, we have developed an automatic system to record newborn cry and movements, during the first six months of life with a specific recording protocol. In this work we present first results of acoustic cry analysis in newborns classified as high-risk subjects being siblings of children already diagnosed as autistics. The work aims at finding possible early ASD signs in high-risk subjects as compared to a group of control subjects based on the fundamental frequency and the vocal tract resonance frequencies. Also, voiced and unvoiced parts of signal and cry-episodes duration are analyzed.

Identification and characterization of novel loss of function mutations in ATP-binding cassette transporter A1 in patients with low plasma high-density lipoprotein cholesterol.

OBJECTIVES: The current literature provides little information on the frequency of mutations in the ATP-binding cassette transporter A1 (ABCA1) in patients with low high-density lipoprotein cholesterol (HDL) levels that are referred to the clinic. In 78 patients with low plasma levels of HDL cholesterol that were referred to our clinic, we routinely screened for ABCA1 gene mutations and studied the functionality of newly identified ABCA1 missense mutations. METHODS: The coding regions and exon-intron boundaries of the ABCA1 gene were sequenced in 78 subjects with HDL cholesterol levels below the 10th percentile for age and gender. Novel mutations were studied by assessing cholesterol efflux capacity (using apolipoprotein A-I as acceptor) after transient expression of ABCA1 variants in BHK cells. RESULTS: Sixteen out of 78 patients (21%) were found to carry 19 different ABCA1 gene variants (1 frameshift, 2 splice-site, 4 nonsense and 12 missense variation) of which 14 variations were novel. Of three patients with homozygous mutations and three patients having compound heterozygous mutations only one patient presented with the clinical characteristics of Tangier Disease (TD) in the presence of nearly complete HDL deficiency. Seven out of eight newly identified ABCA1 missense mutations were found to exhibit a statistically significant loss of cholesterol efflux capacity. CONCLUSION: This study shows that one out of five patients who are referred to our hospital because of low HDL cholesterol levels have a functional ABCA1 gene mutation. It is furthermore demonstrated that in vitro studies are needed to assess functionality of ABCA1 missense mutations.

Recovery of 0.1Hz microvascular skin blood flow in dysautonomic diabetic (type 2) neuropathy by using Frequency Rhythmic Electrical Modulation System (FREMS).

Synchronized oscillation of smooth muscle cells tension in arterioles is the main control system of microvascular skin blood flow. An important autogenic vasomotion activity is recognized in 0.1Hz oscillations through power spectrum analysis of laser Doppler flowmetry. Severe dysautonomia in diabetic neuropathy is correlated with loss of 0.1Hz vasomotor activity, hence with impaired blood microcirculation. FREMS is a novel transcutaneous electrotherapy characterized by sequences of electrical stimuli of high voltage and low pulse duration which vary both in frequency and duration. We have evaluated the changes in laser Doppler flow in the volar part of the forearm before, during and after FREMS. Normal controls (n=10, 6 females, age range 21-39 years) demonstrated significant 0.1Hz vasomotion power spectra at baseline conditions associated with large oscillations of adrenergic cutaneous sweat activity sampled from the hand; people with diabetes type 2 and severe dysautonomia (n=10, 5 females, age range 63-75 years) displayed a significant decrease of 0.1Hz vasomotion power spectra. During FREMS application we observed an increase (p<0.05) of 0.1Hz vasomotion power spectra only in the diabetic group, despite persistence of adrenergic cutaneous sweat activity suppression in this group. However, after the application of the stimuli, the relative energy values around the 0.1Hz peak remained significantly higher than preapplication values in the diabetic group (p<0.05). From these findings, we suggest that FREMS is able to synchronize smooth cell activity, inducing and increasing 0.1Hz vasomotion, independently from the autonomic nervous system.

Severe HDL deficiency due to novel defects in the ABCA1 transporter.

OBJECTIVES: The objective was the identification and functional characterization of mutations in the ABCA1 gene in four patients with severe HDL deficiency. SUBJECTS: Patients were referred to the clinic because of almost complete HDL deficiency. METHODS: The ABCA1 gene was sequenced directly. The analysis of the ABCA1 protein, ABCA1 mRNA and ABCA1-mediated cholesterol efflux was performed in cultured fibroblasts. Intracellular localization of ABCA1 mutants was investigated in transfected HEK293 cells. RESULTS: Two patients were homozygous for mutations in the coding region of the ABCA1 gene, resulting in an amino acid substitution (p.A1046D) and a truncated protein (p.I74YFsX76). The third patient was homozygous for a splice site mutation in intron 35 (c.4773 + 1g>a), resulting in an in-frame deletion of 25 amino acids (del p.D1567_K1591) in ABCA1. These patients had clinical manifestations of accumulation of cholesterol in the reticulo-endothelial system. The fourth patient, with preclinical atherosclerosis, was a compound heterozygote for two missense mutations (p.R587W/p.W1699C). ABCA1-mediated cholesterol efflux was abolished in fibroblasts from patients with p.A1046D and del p.D1567_K1591 mutants and in fibroblasts homozygous for p.R587W. A reduced ABCA1 protein content was observed in these cells, suggesting an increased intracellular degradation. The mutant p.W1699C was largely retained in the endoplasmic reticulum, when expressed in HEK293 cells. CONCLUSIONS: The homozygotes for mutations which abolish ABCA1 function showed overt signs of involvement of the reticulo-endothelial system. This was not the case in the compound heterozygote for missense mutations, suggesting that this patient retains some residual ABCA1 function that reduces cholesterol accumulation in the reticulo-endothelial system.

High-resolution cry analysis in preterm newborn infants.

Infant monitoring is a common procedure in clinical practice in neonatal critical care units. A number of vital functions are monitored, such as heart beat, breathing, blood flow, etc. Specifically, preterm and/or low-birth-weight infants often present respiratory problems that require monitoring. These may range from insufficient ventilation to apnoea. One of the most common events that may affect the respiratory flow is crying, a physiological action made by the infant to communicate and draw attention, but, for a preterm infant, this action requires great effort, which may cause distress and even may have an adverse impact on blood oxygenation. Acoustic analysis of newborn infant cry is thus of importance, since it is related to other basic neuro-physiological parameters. Being easy to perform, cheap and completely non-invasive, it can be successfully applied in many circumstances. The newborn infant cry is characterised by very high fundamental frequency (F(0)) and resonance frequency (RFs) values, with abrupt changes and voiced/unvoiced features of very short duration in a single utterance. To deal with such signals, a new user-friendly software tool has been developed, that allows robust tracking of main acoustic parameters on very short and time-varying signal frames. The software developed provides the user with a high-resolution picture of the cry signal characteristics.

A comparison between internal and surface temperature measurement techniques during phacoemulsification cataract surgery: thermocamera versus thermocouple.

PURPOSE: Corneal and scleral burns, one of the main complications that can occur during a cataract operation, are produced by overheating due to the use of the phacoemulsifier. The temperature of the anterior chamber of the eye can be measured both invasively using thermocouples and non-invasively, but only superficially, using a thermocamera. METHODS: To compare the measures obtained from both techniques an in vitro experimental analysis was conducted on pigs' eyes. During a simulated phacoemulsification cataract operation both the surface temperature with a thermocamera and the temperature inside the anterior chamber with a thermocouple were recorded. For each procedure, the maximum temperature values measured by each technique were compared. RESULTS: The results of this research show that the difference between the maximum values measured with the two techniques is on average 0.5 degrees C. CONCLUSIONS: It is possible to employ a thermocamera technique instead of a thermocouple technique to provide an indication of the temperature inside the anterior chamber.

Non-invasive distress evaluation in preterm newborn infants.

With the increased survival of very preterm infants, there is a growing concern for their developmental outcomes. Infant cry characteristics reflect the development and possibly the integrity of the central nervous system. In this paper, relationships between fundamental frequency (F(0)) and vocal tract resonance frequencies (F(1)-F(3)) are investigated for a set of preterm newborns, by means of a multi-purpose voice analysis tool (BioVoice), characterised by high-resolution and tracking capabilities. Also, first results about possible distress occurring during cry in preterm newborn infants, as related to the decrease of central blood oxygenation, are presented. To this aim, a recording system (Newborn Recorder) has been developed, that allows synchronised, non-invasive monitoring of blood oxygenation and audio recordings of newborn infant's cry. The method has been applied to preterm newborns at the Intensive Care Unit, A.Meyer Children Hospital, Firenze, Italy.

Characterization of the slowly inactivating sodium current INa2 in canine cardiac single Purkinje cells.

The aim of our experiments was to investigate by means of a whole cell patch-clamp technique the characteristics of the slowly inactivating sodium current (I(Na2)) found in the plateau range in canine cardiac Purkinje single cells. The I(Na2) was separated from the fast-activating and -inactivating I(Na) (labelled here I(Na1)) by applying a two-step protocol. The first step, from a holding potential (V(h)) of -90 or -80 mV to -50 mV, led to the quick activation and inactivation of I(Na1). The second step consisted of depolarizations of increasing amplitude from -50 mV to less negative values, which led to the quick activation and slow inactivation of I(Na2). The I(Na2) was fitted with a double exponential function with time constants of tens and hundreds milliseconds, respectively. After the activation and inactivation of I(Na1) at -50 mV, the slope conductance was very small and did not change with time. Instead, during I(Na2), the slope conductance was larger and decreased as a function of time. Progressively longer conditioning steps at -50 mV resulted in a progressive decrease in amplitude of I(Na2) during the subsequent test steps. Gradually longer hyperpolarizing steps (increments of 100 ms up to 600 ms) from V(h) -30 mV to -100 mV were followed on return to -30 mV by a progressively larger I(Na2), as were gradually more negative 500 ms steps from V(h) -30 mV to -90 mV. At the end of a ramp to -20 mV, a sudden repolarization to approximately -35 mV fully deactivated I(Na2). The I(Na2) was markedly reduced by lignocaine (lidocaine) and by low extracellular [Na(+)], but it was little affected by low and high extracellular [Ca(2+)]. At negative potentials, the results indicate that there was little overlap between I(Na2) and the transient outward current, I(to), as well as the calcium current, I(Ca). In the absence of I(to) and I(Ca) (blocked by means of 4-aminopyridine and nickel, respectively), I(Na2) reversed at 60 mV. In conclusion, I(Na2) is a sodium current that can be initiated after the inactivation of I(Na1) and has characteristics that are quite distinct from those of I(Na1). The results have a bearing on the mechanisms underlying the long plateau of Purkinje cell action potential and its modifications in different physiological and pathological conditions.

Shape analysis of microcalcifications using Radon transform.

Microcalcifications are one of the early signs of breast cancer, and they are of great importance for an early diagnosis. Moreover, the spatial distribution and the shape of the microcalcifications have a significant impact in medical practice to evaluate the probability of malignancy of the tumor. In this work a method, performing computer-aided classification of the shape of calcifications accordingly to the classification scheme proposed by Le Gal, is presented. In the first stage, in order to remove mammographic background, the image is preprocessed with a matched filter, designed by modeling the microcalcifications as Gaussian spots and the image as a Fractional Brownian Motion. Afterwards, morphology of spots has been evaluated using two different sets of parameters. The first set utilizes the moments of inertia of the second and third order to compute a set of features, which are invariant to rotations and translations of the image. The second set of parameters is derived from the evaluation of the Radon transform, as computed along eight axes. The results of the Radon transform are used to associate to each lesion a set of features, which are invariant to rotation and scaling of the image. In the final stage, a multilayer neural network has been used to assign each microcalcification to the classes introduced by Le Gal. The topology of the neural network is the same for both sets of descriptors, in order to allow comparison of the discriminative power of the two feature sets. Experimental results obtained with the proposed method from a set of digitized mammograms are reported and discussed.