Wireless Dynamic Light Scattering Sensors Detect Microvascular Changes Associated With Ageing and Diabetes.

This article presents clinical results of wireless portable dynamic light scattering sensors that implement laser Doppler flowmetry signal processing. It has been verified that the technology can detect microvascular changes associated with diabetes and ageing in volunteers. Studies were conducted primarily on wrist skin. Wavelet continuous spectrum calculation was used to analyse the obtained time series of blood perfusion recordings with respect to the main physiological frequency ranges of vasomotions. In patients with type 2 diabetes, the area under the continuous wavelet spectrum in the endothelial, neurogenic, myogenic, and cardio frequency ranges showed significant diagnostic value for the identification of microvascular changes. Aside from spectral analysis, autocorrelation parameters were also calculated for microcirculatory blood flow oscillations. The groups of elderly volunteers and patients with type 2 diabetes, in comparison with the control group of younger healthy volunteers, showed a statistically significant decrease of the normalised autocorrelation function in time scales up to 10 s. A set of identified parameters was used to test machine learning algorithms to classify the studied groups of young controls, elderly controls, and diabetic patients. Our conclusion describes and discusses the classification metrics that were found to be most effective.

Nrf2 regulates glucose uptake and metabolism in neurons and astrocytes.

The transcription factor Nrf2 and its repressor Keap1 mediate cell stress adaptation by inducing expression of genes regulating cellular detoxification, antioxidant defence and energy metabolism. Energy production and antioxidant defence employ NADH and NADPH respectively as essential metabolic cofactors; both are generated in distinct pathways of glucose metabolism, and both pathways are enhanced by Nrf2 activation. Here, we examined the role of Nrf2 on glucose distribution and the interrelation between NADH production in energy metabolism and NADPH homeostasis using glio-neuronal cultures isolated from wild-type, Nrf2-knockout and Keap1-knockdown mice. Employing advanced microscopy imaging of single live cells, including multiphoton fluorescence lifetime imaging microscopy (FLIM) to discriminate between NADH and NADPH, we found that Nrf2 activation increases glucose uptake into neurons and astrocytes. Glucose consumption is prioritized in brain cells for mitochondrial NADH and energy production, with a smaller contribution to NADPH synthesis in the pentose phosphate pathway for redox reactions. As Nrf2 is suppressed during neuronal development, this strategy leaves neurons reliant on astrocytic Nrf2 to maintain redox balance and energy homeostasis.

Diagnosis of Skin Vascular Complications Revealed by Time-Frequency Analysis and Laser Doppler Spectrum Decomposition.

Nowadays, photonics-based techniques are used extensively in various applications, including functional clinical diagnosis, progress monitoring in treatment, and provision of metrological control. In fact, in the frame of practical implementation of optical methods, such as laser Doppler flowmetry (LDF), the qualitative interpretation and quantitative assessment of the detected signal remains vital and urgently required. In the conventional LDF approach, the key measured parameters, index of microcirculation and perfusion rate, are proportional to an averaged concentration of red blood cells (RBC) and their average velocity within a diagnostic volume. These quantities compose mixed signals from different vascular beds with a range of blood flow velocities and are typically expressed in relative units. In the current paper we introduce a new signal processing approach for the decomposition of LDF power spectra in terms of ranging blood flow distribution by frequency series. The developed approach was validated in standard occlusion tests conducted on healthy volunteers, and applied to investigate the influence of local pressure rendered by a probe on the surface of the skin. Finally, in limited clinical trials, we demonstrate that the approach can significantly improve the diagnostic accuracy of detection of microvascular changes in the skin of the feet in patients with Diabetes Mellitus type 2, as well as age-specific changes. The results obtained show that the developed approach of LDF signal decomposition provides essential new information about blood flow and blood microcirculation and has great potential in the diagnosis of vascular complications associated with various diseases.

Brain region specificity in reactive oxygen species production and maintenance of redox balance.

The brain produces various reactive oxygen species in enzymatic and non-enzymatic reactions as a by-product of metabolism and/or for redox signaling. Effective antioxidant system in the brain cells maintains redox balance. However, neurons and glia from some brain regions are more vulnerable to oxidative stress in ischemia/reperfusion, epilepsy, and neurodegenerative disorders than the rest of the brain. Using fluorescent indicators in live cell imaging and confocal microscopy, we have measured the rate of cytosolic and mitochondrial reactive oxygen species production, lipid peroxidation, and glutathione levels in cortex, hippocampus, midbrain, brain stem and cerebellum in acute slices of rat brain. We have found that the basal rate of ROS production is at its highest in brain stem and cerebellum, and that it is mainly generated by glial cells. Activation of neurons and glia by glutamate and ATP led to maximal rates of ROS production in the midbrain compared to the rest of the brain. Mitochondrial ROS had only minor implication to the total ROS production with maximal values in the cortex and minimal in the midbrain. The basal rate of lipid peroxidation was higher in the midbrain and hippocampus, while the GSH level was similar in most brain regions with the lowest level in the midbrain. Thus, the rate of ROS production, lipid peroxidation and the level of GSH vary across brain regions.

Body Position Affects Capillary Blood Flow Regulation Measured with Wearable Blood Flow Sensors.

In this study we demonstrate what kind of relative alterations can be expected in average perfusion and blood flow oscillations during postural changes being measured in the skin of limbs and on the brow of the forehead by wearable laser Doppler flowmetry (LDF) sensors. The aims of the study were to evaluate the dynamics of cutaneous blood perfusion and the regulatory mechanisms of blood microcirculation in the areas of interest, and evaluate the possible significance of those effects for the diagnostics based on blood perfusion monitoring. The study involved 10 conditionally healthy volunteers (44 ± 12 years). Wearable laser Doppler flowmetry monitors were fixed at six points on the body: two devices were fixed on the forehead, on the brow; two were on the distal thirds of the right and left forearms; and two were on the distal thirds of the right and left lower legs. The protocol was used to record three body positions on the tilt table for orthostatic test for each volunteer in the following sequence: (a) supine body position; (b) upright body position (+75°); (c) tilted with the feet elevated above the head and the inclination of body axis of 15° (-15°, Trendelenburg position). Skin blood perfusion was recorded for 10 min in each body position, followed by the amplitude-frequency analysis of the registered signals using wavelet decomposition. The measurements were supplemented with the blood pressure and heart rate for every body position analysed. The results identified a statistically significant transformation in microcirculation parameters of the average level of skin blood perfusion and oscillations of amplitudes of neurogenic, myogenic and cardiac sensors caused by the postural changes. In paper, we present the analysis of microcirculation in the skin of the forehead, which for the first time was carried out in various positions of the body. The area is supplied by the internal carotid artery system and can be of particular interest for evaluation of the sufficiency of blood supply for the brain.

Two-photon conversion of a bacterial phytochrome.

In nature, sensory photoreceptors underlie diverse spatiotemporally precise and generally reversible biological responses to light. Photoreceptors also serve as genetically encoded agents in optogenetics to control by light organismal state and behavior. Phytochromes represent a superfamily of photoreceptors that transition between states absorbing red light (Pr) and far-red light (Pfr), thus expanding the spectral range of optogenetics to the near-infrared range. Although light of these colors exhibits superior penetration of soft tissue, the transmission through bone and skull is poor. To overcome this fundamental challenge, we explore the activation of a bacterial phytochrome by a femtosecond laser emitting in the 1 µm wavelength range. Quantum chemical calculations predict that bacterial phytochromes possess substantial two-photon absorption cross sections. In line with this notion, we demonstrate that the photoreversible Pr ↔ Pfr conversion is driven by two-photon absorption at wavelengths between 1170 and 1450 nm. The Pfr yield was highest for wavelengths between 1170 and 1280 nm and rapidly plummeted beyond 1300 nm. By combining two-photon activation with bacterial phytochromes, we lay the foundation for enhanced spatial resolution in optogenetics and unprecedented penetration through bone, skull, and soft tissue.

Laser Doppler Spectrum Analysis Based on Calculation of Cumulative Sums Detects Changes in Skin Capillary Blood Flow in Type 2 Diabetes Melitus.

In this article, we introduce a new method of signal processing and data analysis for the digital laser Doppler flowmetry. Our approach is based on the calculation of cumulative sums over the registered Doppler power spectra. The introduced new parameter represents an integral estimation for the redistribution of moving red blood cells over the range of speed. The prototype of the device implementing the technique is developed and tested in preliminary clinical trials. The methodology was verified with the involvement of two age groups of healthy volunteers and in a group of patients with type 2 diabetes mellitus. The main practical result of the study is the development of a set of binary linear classifiers that allow the method to identify typical patterns of the microcirculation for the healthy volunteers and diabetic patients based on the presented diagnostic algorithm.

Skin Complications of Diabetes Mellitus Revealed by Polarized Hyperspectral Imaging and Machine Learning.

Aging and diabetes lead to protein glycation and cause dysfunction of collagen-containing tissues. The accompanying structural and functional changes of collagen significantly contribute to the development of various pathological malformations affecting the skin, blood vessels, and nerves, causing a number of complications, increasing disability risks and threat to life. In fact, no methods of non-invasive assessment of glycation and associated metabolic processes in biotissues or prediction of possible skin complications, e.g., ulcers, currently exist for endocrinologists and clinical diagnosis. In this publication, utilizing emerging photonics-based technology, innovative solutions in machine learning, and definitive physiological characteristics, we introduce a diagnostic approach capable of evaluating the skin complications of diabetes mellitus at the very earlier stage. The results of the feasibility studies, as well as the actual tests on patients with diabetes and healthy volunteers, clearly show the ability of the approach to differentiate diabetic and control groups. Furthermore, the developed in-house polarization-based hyperspectral imaging technique accomplished with the implementation of the artificial neural network provides new horizons in the study and diagnosis of age-related diseases.

Machine Learning Aided Photonic Diagnostic System for Minimally Invasive Optically Guided Surgery in the Hepatoduodenal Area.

Abdominal cancer is a widely prevalent group of tumours with a high level of mortality if diagnosed at a late stage. Although the cancer death rates have in general declined over the past few decades, the mortality from tumours in the hepatoduodenal area has significantly increased in recent years. The broader use of minimal access surgery (MAS) for diagnostics and treatment can significantly improve the survival rate and quality of life of patients after surgery. This work aims to develop and characterise an appropriate technical implementation for tissue endogenous fluorescence (TEF) and assess the efficiency of machine learning methods for the real-time diagnosis of tumours in the hepatoduodenal area. In this paper, we present the results of the machine learning approach applied to the optically guided MAS. We have elaborated tissue fluorescence approach with a fibre-optic probe to record the TEF and blood perfusion parameters during MAS in patients with cancers in the hepatoduodenal area. The measurements from the laser Doppler flowmetry (LDF) channel were used as a sensor of the tissue vitality to reduce variability in TEF data. Also, we evaluated how the blood perfusion oscillations are changed in the tumour tissue. The evaluated amplitudes of the cardiac (0.6-1.6 Hz) and respiratory (0.2-0.6 Hz) oscillations was significantly higher in intact tissues (p < 0.001) compared to the cancerous ones, while the myogenic (0.2-0.06 Hz) oscillation did not demonstrate any statistically significant difference. Our results demonstrate that a fibre-optic TEF probe accompanied with ML algorithms such as k-Nearest Neighbours or AdaBoost is highly promising for the real-time in situ differentiation between cancerous and healthy tissues by detecting the information about the tissue type that is encoded in the fluorescence spectrum. Also, we show that the detection can be supplemented and enhanced by parallel collection and classification of blood perfusion oscillations.

Optical percutaneous needle biopsy of the liver: a pilot animal and clinical study.

This paper presents the results of the experiments which were performed using the optical biopsy system specially developed for in vivo tissue classification during the percutaneous needle biopsy (PNB) of the liver. The proposed system includes an optical probe of small diameter acceptable for use in the PNB of the liver. The results of the feasibility studies and actual tests on laboratory mice with inoculated hepatocellular carcinoma and in clinical conditions on patients with liver tumors are presented and discussed. Monte Carlo simulations were carried out to assess the diagnostic volume and to trace the sensing depth. Fluorescence and diffuse reflectance spectroscopy measurements were used to monitor metabolic and morphological changes in tissues. The tissue oxygen saturation was evaluated using a recently developed approach to neural network fitting of diffuse reflectance spectra. The Support Vector Machine Classification was applied to identify intact liver and tumor tissues. Analysis of the obtained results shows the high sensitivity and specificity of the proposed multimodal method. This approach allows to obtain information before the tissue sample is taken, which makes it possible to significantly reduce the number of false-negative biopsies.

Adrenaline induces calcium signal in astrocytes and vasoconstriction via activation of monoamine oxidase.

Adrenaline or epinephrine is a hormone playing an important role in physiology. It is produced de-novo in the brain in very small amounts compared to other catecholamines, including noradrenaline. Although the effects of adrenaline on neurons have been extensively studied, much less is known about the action of this hormone on astrocytes. Here, we studied the effects of adrenaline on astrocytes in primary co-culture of neurons and astrocytes. Application of adrenaline induced calcium signal in both neurons and astrocytes, but only in neurons this effect was dependent on α- and ß-receptor antagonists. The effects of adrenaline on astrocytes were less dependent on adrenoreceptors: the antagonist carvedilol had only moderate effect on the calcium signal and the agonist of adrenoreceptors methoxamine induced a signal only in small proportion of the cells. We found that adrenaline in astrocytes activates phospholipase C and subsequent release of calcium from the endoplasmic reticulum. Calcium signal in astrocytes is initiated by the metabolism of adrenaline by the monoamine oxidase (MAO), which activates reactive oxygen species production and induces lipid peroxidation. Inhibitor of MAO selegiline inhibited both adrenaline-induced calcium signal in astrocytes and the vasoconstriction that indicates an important role for monoamine oxidase in adrenaline-induced signalling and function.

Biophotonics methods for functional monitoring of complications of diabetes mellitus.

The prevalence of diabetes complications is a significant public health problem with a considerable economic cost. Thus, the timely diagnosis of complications and prevention of their development will contribute to increasing the length and quality of patient life, and reducing the economic costs of their treatment. This article aims to review the current state-of-the-art biophotonics technologies used to identify the complications of diabetes mellitus and assess the quality of their treatment. Additionally, these technologies assess the structural and functional properties of biological tissues, and they include capillaroscopy, laser Doppler flowmetry and hyperspectral imaging, laser speckle contrast imaging, diffuse reflectance spectroscopy and imaging, fluorescence spectroscopy and imaging, optical coherence tomography, optoacoustic imaging and confocal microscopy. Recent advances in the field of optical noninvasive diagnosis suggest a wider introduction of biophotonics technologies into clinical practice and, in particular, in diabetes care units.

A complex morphofunctional approach for zinc toxicity evaluation in rats.

Anthropogenic activity causes the introduction of zinc compounds into the biological cycle in mining and processing sites and its accumulation in organs and tissues, causing systemic toxicity. A cumulative effect of zinc is predominantly neurotoxic and it also affects the respiratory, cardiovascular and digestive systems. This study evaluates the effects of single-dose intragastric administration of 100 mg/kg zinc succinate on the structure and function of organs and tissues in male Wistar rats 1 month after treatment. The presented morphofunctional approach for the toxicity evaluation included the study of behavioral responses using the automated Laboras® complex, fluorescent spectral analysis of the NADH and FAD activity and histological evaluation of animal organs and tissues. The results of the behavioral activity assessment showed a significant decrease in animals' motor activity, whereas the fluorescence spectra analysis demonstrated a decrease in coenzyme NADH without the reduction of FAD levels. We detected toxic and dystrophic changes in the cerebral cortex, heart, lungs and liver tissues. Our original multiparametric approach enables a comprehensive assessment of the long-term toxic effects of the metal salts such as zinc succinate, especially in the cerebral cortex at the doses much lower than the acute LD50 reported for the common zinc salts.

Wavelet Analysis of the Temporal Dynamics of the Laser Speckle Contrast in Human Skin.

OBJECTIVE: Spectral analysis of laser Doppler flowmetry (LDF) signals has been widely used in studies of physiological vascular function regulation. An alternative to LDF is the laser speckle contrast imaging method (LSCI), which is based on the same physical principle. In contrast to LDF, LSCI provides non-scanning full-field imaging of a relatively wide skin area and offers high spatial and temporal resolutions, which allows visualization of microvascular structure. This circumstance, together with a large number of works which had shown the effectiveness of temporal LSCI analysis, gave impetus to experimental studies of the relation between LDF and LSCI used to monitor the temporal dynamics of blood flow. METHODS: Continuous wavelet transform was applied to construct a time-frequency representation of a signal. RESULTS: Analysis of 10 minute LDF and LSCI output signals recorded simultaneously revealed rather high correlation between oscillating components. It was demonstrated for the first time that the spectral energy of oscillations in the 0.01-2 Hz frequency range of temporal LSCI recordings carries the same information as the conventional LDF recordings and hence it reflects the same physiological vascular tone regulation mechanisms. CONCLUSION: The approach proposed can be used to investigate speckle pattern dynamics by LSCI in both normal and pathological conditions. SIGNIFICANCE: The results of research on the influence of spatial binning and averaging on the spectral characteristics of perfusion monitored by LSCI are of considerable interest for the development of LSCI systems optimized to evaluate temporal dynamics.

Influence of blood pulsation on diagnostic volume in pulse oximetry and photoplethysmography measurements.

Recent advances in the development of ultra-compact semiconductor lasers and technology of printed flexible hybrid electronics have opened broad perspectives for the design of new pulse oximetry and photoplethysmography devices. Conceptual design of optical diagnostic devices requires careful selection of various technical parameters, including spectral range; polarization and intensity of incident light; actual size, geometry, and sensitivity of the detector; and mutual position of the source and detector on the surface of skin. In the current study utilizing a unified Monte Carlo computational tool, we explore the variations in diagnostic volume due to arterial blood pulsation for typical transmitted and back-scattered probing configurations in a human finger. The results of computational studies show that the variations in diagnostic volumes due to arterial pulse wave are notably (up to 45%) different in visible and near-infrared spectral ranges in both transmitted and back-scattered probing geometries. While these variations are acceptable for relative measurements in pulse oximetry and/or photoplethysmography, for absolute measurements, an alignment normalization of diagnostic volume is required and can be done by a computational approach utilized in the framework of the current study.

Influence of blood pulsation on diagnostic volume in pulse oximetry and photoplethysmography measurements: publisher's note.

This publisher's note corrects several errors in Appl. Opt.58, 9398 (2019).APOPAI0003-693510.1364/AO.58.009398.

Hyperspectral imaging of human skin aided by artificial neural networks.

We developed a compact, hand-held hyperspectral imaging system for 2D neural network-based visualization of skin chromophores and blood oxygenation. State-of-the-art micro-optic multichannel matrix sensor combined with the tunable Fabry-Perot micro interferometer enables a portable diagnostic device sensitive to the changes of the oxygen saturation as well as the variations of blood volume fraction of human skin. Generalized object-oriented Monte Carlo model is used extensively for the training of an artificial neural network utilized for the hyperspectral image processing. In addition, the results are verified and validated via actual experiments with tissue phantoms and human skin in vivo. The proposed approach enables a tool combining both the speed of an artificial neural network processing and the accuracy and flexibility of advanced Monte Carlo modeling. Finally, the results of the feasibility studies and the experimental tests on biotissue phantoms and healthy volunteers are presented.

Multimodal Optical Diagnostics of the Microhaemodynamics in Upper and Lower Limbs.

The introduction of optical non-invasive diagnostic methods into clinical practice can substantially advance in the detection of early microcirculatory disorders in patients with different diseases. This paper is devoted to the development and application of the optical non-invasive diagnostic approach for the detection and evaluation of the severity of microcirculatory and metabolic disorders in rheumatic diseases and diabetes mellitus. The proposed methods include the joint use of laser Doppler flowmetry, absorption spectroscopy and fluorescence spectroscopy in combination with functional tests. This technique showed the high diagnostic importance for the detection of disturbances in peripheral microhaemodynamics. These methods have been successfully tested as additional diagnostic techniques in the field of rheumatology and endocrinology. The sensitivity and specificity of the proposed diagnostic procedures have been evaluated.

Optical probe pressure effects on cutaneous blood flow.

The variation of blood flow characteristics caused by the probe pressure during noninvasive studies is of particular interest within the context of fundamental and applied research. It has been shown previously that the weak local pressure induces vasodilation, whereas the increased pressure is able to stop the blood flow in the compressed area, as well as to significantly change optical signals.The blood flow oscillations measured by laser Doppler flowmetry (LDF) characterize the functional state of the microvascular system and can be used for noninvasive diagnostics of its abnormality. This study was intended to identify the patterns of the relationship between the oscillating components of blood flow registered by the LDF method under different levels of pressure applied to an optical fiber probe.For this purpose, we have developed an original optical probe capable of regulating the applied pressure. The developed protocol included six sequential records of the blood perfusion at a pressure within the 0 to 200 mmHg range with unloading at the last stage.Using wavelet analyses, we traced the variation of energy of oscillations for these records in five frequency bands associated with different vascular tone regulation mechanisms. Six young volunteers of the same age (three males and three females) were included in this preliminary study and the protocol was repeated five times in each volunteer. Accordingly, 30 LDF records were available for the analyses. As expected, the LDF signal increases at weak pressure (30 mmHg) and decreases at increased pressure. The statistically stable amplification of endothelial associated blood flow oscillations under the 90 mmHg pressure allowed us to put forward a hypothesis that the endothelial activity increases. The possible causes of this phenomenon are discussed.

Dynamic evaluation of blood flow microcirculation by combined use of the laser Doppler flowmetry and high-speed videocapillaroscopy methods.

The dynamic light scattering methods are widely used in biomedical diagnostics involving evaluation of blood flow. However, there exist some difficulties in quantitative interpretation of backscattered light signals from the viewpoint of diagnostic information. This study considers the application of the high-speed videocapillaroscopy (VCS) method that provides the direct measurement of the red blood cells (RBCs) velocity into a capillary. The VCS signal presents true oscillation nature of backscattered light caused by moving RBCs. Thus, the VCS signal can be assigned as a reference one with respect to more complicated signals like in laser Doppler flowmetry (LDF). An essential correlation between blood flow velocity oscillations in a separate human capillary and the integral perfusion estimate obtained by the LDF method has been found. The observation of blood flow by the VCS method during upper arm occlusion has shown emergence of the reverse blood flow effect in capillaries that corresponds to the biological zero signal in the LDF. The reverse blood flow effect has to be taken into account in interpretation of LDF signals.