Evgeny A. Zherebtsov

Individual variability analysis of fluorescence parameters measured in skin with different levels of nutritive blood flow

Fluorescence spectroscopy has recently become more common in clinical medicine. However, there are still many unresolved issues related to the methodology and implementation of instruments with this technology. In this study, we aimed to assess individual variability of fluorescence parameters of endogenous markers (NADH, FAD, etc.) measured by fluorescent spectroscopy (FS) in situ and to analyse the factors that lead to a significant scatter of results. Most studied fluorophores have an acceptable scatter of values (mostly up to 30%) for diagnostic purposes. Here we provide evidence that the level of blood volume in tissue impacts FS data with a significant inverse correlation. The distribution function of the fluorescence intensity and the fluorescent contrast coefficient values are a function of the normal distribution for most of the studied fluorophores and the redox ratio. The effects of various physiological (different content of skin melanin) and technical (characteristics of optical filters) factors on the measurement results were additionally studied. The data on the variability of the measurement results in FS should be considered when interpreting the diagnostic parameters, as well as when developing new algorithms for data processing and FS devices.

Substantiation of medical and technical requirements for noninvasive spectrophotometric diagnostic devices

Abstract. A scientific approach to the formulation of medical and technical requirements (MTRs) for noninvasive spectrophotometric diagnostic devices using optical technologies such as laser Doppler flowmetry and absorption spectroscopy is proposed. The theoretical modeling framework, metrological certification, and testing of these devices are still in the early stages of development. The theoretical estimation of the received signal levels for wavelengths between 514 and 940 nm is highly dependent on the blood volume level in the subject tissue. The proposed approach allows, in particular, the calculation of technical and metrological performance constraints of the instruments, such as the ranges of the sensitivity and power-related signal-to-noise ratios for different spectral channels and different biomedical (biochemical and physiological) parameters. Substantiation of specialized MTRs for the noninvasive spectrophotometric diagnostic devices can enable them to develop to the level of standardized measurement techniques.

The development of attenuation compensation models of fluorescence spectroscopy signals

This study examines the effect of blood absorption on the endogenous fluorescence signal intensity of biological tissues. Experimental studies were conducted to identify these effects. To register the fluorescence intensity, the fluorescence spectroscopy method was employed. The intensity of the blood flow was measured by laser Doppler flowmetry. We proposed one possible implementation of the Monte Carlo method for the theoretical analysis of the effect of blood on the fluorescence signals. The simulation is constructed as a four-layer skin optical model based on the known optical parameters of the skin with different levels of blood supply. With the help of the simulation, we demonstrate how the level of blood supply can affect the appearance of the fluorescence spectra. In addition, to describe the properties of biological tissue, which may affect the fluorescence spectra, we turned to the method of diffuse reflectance spectroscopy (DRS). Using the spectral data provided by the DRS, the tissue attenuation effect can be extracted and used to correct the fluorescence spectra.

Study of the functional state of peripheral vessels in fingers of rheumatological patients by means of laser Doppler flowmetry and cutaneous thermometry measurements

Vasospastic disorders are a common class of rheumatic disease. These include syndromes such as vegetative dystonia, Raynauds syndrome, vibration disease and rheumatoid arthritis among others. The aim of this work is to develop an original method of diagnosing the functional state of peripheral vessels of the fingers, based on the simultaneous recording of LDF- and thermograms during the occlusion test, for determining vascular disorders of rheumatological patients. A diagnostic method was developed for assessing the functional state of the peripheral vessels of fingers, based on carrying out occlusion test in a thermally stabilized environment, with simultaneous recording of signals of laser Doppler flowmetry and skin thermometry. To verify the diagnostic value of the proposed method, a series of experiments were carried out on 41 rheumatological patients: 5 male and 36 females (average age 56.0±12.2 years). The most common diagnoses in the patient group were rheumatoid arthritis, arthrosis, gout and systemic lupus erythematosus. The laser analyser of blood microcirculation “LAKK-02” (SPE “LAZMA” Ltd, Russia) and a custom developed multi-channel thermometry device for low inertia thermometry were used for experimental measurements. The measurements of cutaneous temperature and the index of microcirculation were performed on the distal phalanx of the third finger of the right hand. Occlusion tests were performed with water baths at 25 and 42 °C and a tonometer cuff with a pressure of 200-220 mmHg for 3 min on the upper arm. The results of experimental studies are presented and interpreted. These data indicate a violation of the blood supply regulation in the form of a pronounced tendency towards microvascular vasoconstriction in the fingers. Thus, the response displaying a tendency toward angiospasm among patients in the rheumatological diseases profile group was observed mainly in the most severe cases (49 % of this group). The prospects of the developed diagnostic method of microcirculatory disorders in rheumatic diseases are evaluated. Thus, cutaneous blood microcirculation and temperature measurements performed together can help in diagnosis of the functional state of peripheral vessels both in a healthy state and when expressing pathology.

The blood perfusion and NADH/FAD content combined analysis in patients with diabetes foot

Skin blood microcirculation and the metabolism activity of tissue were examined on the patients with type 2 diabetes. Laser Doppler flowmetry (LDF) with 1064 nm laser light source and fluorescence spectroscopy (FS) with excitation light of 365 nm and 450 nm have been used to monitor the blood perfusion and the content of coenzymes NADH and FAD. Concluding, the proposed combined LDF and tissue FS approach allows to identify the significant violations in the blood microcirculation and metabolic activity for type 2 diabetes patients.

Novel measure for the calibration of laser Doppler flowmetry devices

The metrological basis for optical non-invasive diagnostic devices is an unresolved issue. A major challenge for laser Doppler flowmetry (LDF) is the need to compare the outputs from individual devices and various manufacturers to identify variations useful in clinical diagnostics. The most common methods for instrument calibration are simulants or phantoms composed of colloids of light-scattering particles which simulate the motion of red blood cells based on Brownian motion. However, such systems have limited accuracy or stability and cannot calibrate for the known rhythmic components of perfusion (0.0095-1.6 Hz). To solve this problem, we propose the design of a novel technique based on the simulation of moving particles using an electromechanical transducer, in which a precision piezoelectric actuator is used (e.g., P-602.8SL with maximum movement less than 1 mm). In this system, Doppler shift is generated in the layered structure of different solid materials with different optical light diffusing properties. This comprises a fixed, light transparent upper plane-parallel plate and an oscillating fluoroplastic (PTFE) disk. Preliminary studies on this experimental setup using the LDF-channel of a “LAKK-M” system demonstrated the detection of the linear portion (0-10 Hz with a maximum signal corresponding to Doppler shift of about 20 kHz) of the LDF-signal from the oscillating frequency of the moving layer. The results suggest the possibility of applying this technique for the calibration of LDF devices.

Development of a space-borne spectrometer to monitor atmospheric ozone

A new compact satellite spectrometer dedicated to monitoring terrestrial atmospheric ozone (ozonometer) is in preparation for the Russian Geophysics Program. Four instruments at four satellites (Ionosphere) are intended to monitor the total ozone content by measuring spectra of scattered solar radiation in nadir. The spectrometer is based on the Rowland scheme with a concave holographic diffraction grating. It covers the near UV and visible range of the spectrum, 300-500xa0nm, with a spectral resolution of ∼0.3u2009u2009nm. At present, a qualification model has been manufactured and tested. We introduce the description of the instrument and the results of laboratory and ground-based atmospheric calibrations. The ozone amount retrieved from atmospheric measurements using the differential optical absorption spectroscopy (DOAS) method is in good agreement with that measured by the collocated Brewer spectrophotometer and ozone monitoring instrument on board the Aura satellite.

Noninvasive control of rhodamine-loaded capsules distribution in vivo

Using fluorescence spectroscopy system with fibre-optical probe, we investigated the dynamics of propagation and circulation in the microcirculatory system of experimental nanocapsules fluorescent-labelled (rhodamine TRITC) nanocapsules. The studies were carried out in clinically healthy Wistar rats. The model animals were divided into control group and group received injections of the nanocapsules. The fluorescent measurements conducted transcutaneously on the thigh surface. The administration of the preparation with the rhodamine concentration of 5 mg/kg of animal weight resulted in twofold increase of fluorescence intensity by reference to the baseline level. As a result of the study, it was concluded that fluorescence spectroscopy can be used for transdermal measurements of the rhodamine-loaded capsules in vivo.

Peculiarities of local blood microcirculation in patients with psoriasis

Local hemodynamic parameters were studied by means of laser Doppler flowmetry in 15 patients with psoriasis in the stationary stage, who have plaques on the inner surface of the forearm. LDF signals recorded at the site of psoriatic lesions of the tissue as well as in the intact tissue at a distance of 1-2 cm from the affected area were analysed. LDF signals were postprocessed by continuous wavelet transform using the Morlet wavelet.

Blood flow oscillations as a signature of microvascular abnormalities

Laser Doppler flowmetry (LDF) was utilized for blood ow measurements. Wavelet analysis was used to identify spectral characteristics of the LDF signal in patients with rheumatic diseases and diabetes mellitus. Baseline measurements were applied for both pathological groups. Blood flow oscillations analyses were performed by means of the wavelet transform. Higher baseline perfusion was observed in both pathological groups in comparison to controls. Differences in the spectral properties between the groups studied were revealed. The results obtained demonstrated that spectral properties of the LDF signal collected in basal conditions may be the signature of microvasculature functional state.