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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.

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.

This study is devoted to the trials of wearable diagnostic system that implements the laser Doppler flowmetry technique to analyse the blood microcirculation. We do preliminary test with involvement of limited group of healthy volunteers of different age and in patients with type 2 diabetes. During the series of measurements, the microcirculation parameters was measured for 10 minutes in the palmar surfaces of the big toes and in the inner sides of the upper thirds of the shins. A statistically significant differences was found in bypass index, nutritive and shunt blood ow in shins between older group of volunteers and patients’ group as well as in shunt blood flow in fingers between younger and older groups of volunteers.

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 β-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.

The aim of the study was to compare the metabolic activity of brain cortex after the acute hypoxia caused by the impairment of breathing or blood circulation. Male Wistar rats were randomized in two groups: impaired breathing and blood circulation failure groups. Fluorescence under 365 and 450 nm excitation and diffuse reflectance intensity at 550–820 nm range were estimated. We found that after long-term hypoxic conditions, notable metabolic changes occur. We suppose that oxygen deficiency causes an activation of the GABA shunt mechanism. In cases of blood circulation failure, fluorescence intensity changes faster than in cases of breathing impairment.

The article is aimed at conducting pilot studies of microcirculation changes in patients with diabetes mellitus during the course of intravenous infusions of alpha lipoic acid. The study was conducted with patients with diabetes during the passage of a standard course of treatment. The change in parameters was compared before the start of the infusion course, during and after the course. A system of wearable laser Doppler monitors was used to conduct the study. The study showed that although the microcirculation index does not undergo significant changes during the treatment process, spectral analysis of the recorded signal has potential applicability. Further studies with a larger sample group are required for a qualitative analysis of the observed effects.

The growing interest in the development of new wearable electronic devices for mobile healthcare provides great opportunities for the development of methods for assessing blood perfusion in this direction. Laser Doppler flowmetry (LDF) is one of the promising methods. A fine analysis of capillary blood ow structure and rhythm in the time and frequency domains, coupled with a new possibility of round-the-clock monitoring can provide valuable diagnostic information about the state of microvascular blood ow. In this study, wearable implementation of laser Doppler flowmetry was utilised for microcirculatory function assessment in patients with diabetes and healthy controls of two distinct age groups. Four wearable laser Doppler flowmetry monitors were used for the analysis of blood microcirculation. Thirty-seven healthy volunteers and 18 patients with type 2 diabetes mellitus participated in the study. The results of the studies have shown that the average perfusion differs between healthy volunteers of distinct age groups and between healthy volunteers of the younger age group and patients with diabetes mellitus. It was noted that the average level of perfusion measured on the wrist in the two groups of healthy volunteers has no statistically significant differences found in similar measurements on the fingertips. The wearable implementation of LDF can become a truly new diagnostic interface to monitor cardiovascular parameters, which could be of interest for diagnostics of conditions associated with microvascular disorders.

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.

The paper presents the results of in vivo experimental measurements of the endogenous fluorescence and blood perfusion in common bile duct tissues in obstructive jaundice patients during minimally invasive surgical operations. Nowadays the type of interventions become more widespread for treating hepatopancreatobiliary pathologies. An effective feedback which can be established by one or more optical diagnostic channels integrated into the standard instruments for minimally invasive manipulations would be of particular value to practical surgeons. To collect reference data, an experimental study has been conducted using a fiber optical probe with fluorescence spectroscopy and laser Doppler flowmetry channels. For every measurement, the areas of interest were different areas of common bile duct obstructed by a malignant tumor. Obtained results demonstrated significant variety registered intensities of fluorescence spectra and values of blood perfusion. Tested fiber optical endoscopic system showed sensitivity to the tissue state in the investigated area, so the two modal approach seems to be promising for further research and implementation in clinical practice.

Presently, in the modern laser Doppler flowmetry (LDF) the distribution of blood perfusion and its changes along the Doppler shift frequencies are simply ignored and/or not properly addressed. Utilizing the registered power spectrum of photocurrent, we introduce an LDF signal processing approach suitable for expanding of diagnostic capabilities of the technique. In particular, we demonstrate that it is possible to determine how the oscillations of blood ow (cardiac, breathe, myogenic, etc.) are distributed along the Doppler shift frequency. Wavelet analysis is utilized to extract the oscillations corresponded to the particular frequency sub-bands of blood perfusion. The main purpose of this study is to identify influence of local pressure by fiber optic probe on cardiac oscillations and their distribution along frequency of Doppler shift.

A novel non-invasive, wearable VCSEL-based system for multipoint in − vivo measurements of blood perfusion was introduced. The system operates on the basis of the laser Doppler flowmetry (LDF) method and allows for microcirculation studies. The sensors developed were used to analyse the skin blood flow synchronization in homologous regions of the contralateral limbs, both in the basal state and during various functional tests. A high synchronisation of blood flow rhythms in the contralateral limbs of healthy volunteers was shown in the studies presented.

Videocapillaroscopy (VCS) and laser Doppler flowmetry (LDF) are non-invasive methods for evaluating microcirculation parameters. The VCS method is based on a high-speed video recording of capillaries in the nailfold. The recorded video frames are processed using a specialized algorithm to determine the red blood cells velocity. The LDF technique is based on the analysis of the Doppler shift of back-scattered laser radiation from moving red blood cells. In this work, simultaneous measurements of VCS and LDF have been performed in healthy volunteers and rheumatic patient. The study was conducted using a cold pressor test. Changes were recorded in response to cold exposure in rheumatic diseases.

We introduce a noninvasive diagnostic approach for functional monitoring of blood microflows in capillaries and thermoregulatory vessels within the skin. The measuring system is based on the combined use of laser Doppler flowmetry and skin contact thermometry. The obtained results suggest that monitoring of blood microcirculation during the occlusion, performed in conjunction with the skin temperature measurements in the thermally stabilized medium, has a great potential for quantitative assessment of angiospatic dysfunctions of the peripheral blood vessels. The indices of blood flow reserve and temperature response were measured and used as the primarily parameters of the functional diagnostics of the peripheral vessels of skin. Utilizing these parameters, a simple phenomenological model has been suggested to identify patients with angiospastic violations in the vascular system.

According to the International Diabetes Federation, the challenge of early stage diagnosis and treatment effectiveness monitoring in diabetes is currently one of the highest priorities in modern healthcare. The potential of combined measurements of skin fluorescence and blood perfusion by the laser Doppler flowmetry method in diagnostics of low limb diabetes complications was evaluated. Using Monte Carlo probabilistic modeling, the diagnostic volume and depth of the diagnosis were evaluated. The experimental study involved 76 patients with type 2 diabetes mellitus. These patients were divided into two groups depending on the degree of complications. The control group consisted of 48 healthy volunteers. The local thermal stimulation was selected as a stimulus on the blood microcirculation system. The experimental studies have shown that diabetic patients have elevated values of normalized fluorescence amplitudes, as well as a lower perfusion response to local heating. In the group of people with diabetes with trophic ulcers, these parameters also significantly differ from the control and diabetes only groups. Thus, the intensity of skin fluorescence and level of tissue blood perfusion can act as markers for various degrees of complications from the beginning of diabetes to the formation of trophic ulcers.