Oxana V. Semyachkina-Glushkovskaya

Multiresolution analysis of pathological changes in cerebral venous dynamics in newborn mice with intracranial hemorrhage: adrenorelated vasorelaxation

Intracranial hemorrhage (ICH) is the major problem of modern neonatal intensive care. Abnormalities of cerebral venous blood flow (CVBF) can play a crucial role in the development of ICH in infants. The mechanisms underlying these pathological processes remain unclear; however it has been established that the activation of the adrenorelated vasorelaxation can be an important reason. Aiming to reach a better understanding of how the adrenodependent relaxation of cerebral veins contributes to the development of ICH in newborns, we study here the effects of pharmacological stimulation of adrenorelated dilation of the sagittal sinus by isoproterenol on the cerebral venous hemodynamics. Our study is performed in newborn mice at different stages of ICH using the laser speckle contrast imaging and wavelet analysis of the vascular dynamics of CVBF. We show that the dilation of the sagittal sinus with the decreased velocity of blood flow presides to the stress-induced ICH in newborn mice. These morphofunctional vascular changes are accompanied by an increased variance of the wavelet-coefficients in the areas of endothelial and non-endothelial (KATP-channels activity of vascular muscle) sympathetic components of the CVBF variability. Changes in the cerebral venous hemodynamics at the latent stage of ICH are associated with a high responsiveness of the sagittal sinus to isoproterenol quantifying by wavelet-coefficients related to a very slow region of the frequency domain. The obtained results certify that a high activation of the adrenergic-related vasodilatory responses to severe stress in newborn mice can be one of the important mechanisms underlying the development of ICH. Thus, the venous insufficiency with the decreased blood outflow from the brain associated with changes in the endothelial and the sympathetic components of CVBF-variability can be treated as prognostic criteria for the risk of ICH during the first days after birth.

THE EXPERIMENTAL STUDY OF STRESS-RELATED PATHOLOGICAL CHANGES IN CEREBRAL VENOUS BLOOD FLOW IN NEWBORN RATS ASSESSED BY DOCT

In experiments on newborn rats with stress-related intracranial hemorrhage (ICH) using Doppler optical coherence tomography (DOCT) we have shown that latent stage of ICH (4 h after stress) is characterized by decrease of venous blood outflow and the loss of sensitivity of sagittal vein to vasoconstrictor effect of adrenaline. The incidence of ICH (24 h after stress) was accompanied by progression of early pathological changes in cerebral venous blood flow (CVBF) and development of venous insufficiency. Taking into consideration of this fact, we suggest that the suppression of CVBF related to the severity to the deleterious effect of stress on the brain hemodynamics in newborn rats. These facts allow us to conclude that the venous insufficiency with the loss of vasoconstrictor response to adrenaline is an informative and sensitive component of pattern of CVBF that can be important diagnostic criteria of risk of ICH development in newborns.

Optical monitoring of stress-related changes in the brain tissues and vessels associated with hemorrhagic stroke in newborn rats

Stress is a major factor for a risk of cerebrovascular catastrophes. Studying of mechanisms underlying stress-related brain-injures in neonates is crucial for development of strategy to prevent of neonatal stroke. Here, using a model of sound-stress-induced intracranial hemorrhages in newborn rats and optical methods, we found that cerebral veins are more sensitive to the deleterious effect of stress than arteries and microvessels. The development of venous insufficiency with decreased blood outflow from the brain accompanied by hypoxia, reduction of complexity of venous blood flow and high production of beta-arrestin-1 are possible mechanisms responsible for a risk of neonatal hemorrhagic stroke.

WAVELET-BASED ANALYSIS OF CEREBROVASCULAR DYNAMICS IN NEWBORN RATS WITH INTRACRANIAL HEMORRHAGES

Intracranial hemorrhage (IH) is a major problem of neonatal intensive care. The incidence of IH is typically asymptomatic and cannot be effectively detected by standard diagnostic methods. The mechanisms underlying IH are unknown but there is evidence that stress-induced disorders in adrenergic regulation of cerebral venous blood flow (CVBF) are among the main reasons. Quantitative and qualitative assessment of CVBF could significantly advance understanding of the nature of IH in newborns. In this work, we analyze variations of CVBF in newborn rats with an experimental model of stress-induced IH and adrenaline injection. Our analysis is based on the Doppler optical coherence tomography (DOCT) and a proposed adaptive wavelet-based approach that provides sensitive markers of abnormal reactions of the sagittal vein to external factors. The obtained results demonstrate that the incidence of IH in newborn rats is accompanied by a suppression of CVBF with the development of venous insufficiency and areactivity to adrenaline. We introduce a numerical measure θ, quantifying reactions of CVBF and show that the values θ < 1.23 estimated in the low-frequency (LF) spectral range corresponding to the sympathicus indicate abnormal reactions associated with the development of IH. We conclude that the revealed areactivity of the cerebral veins to adrenaline represents a possible mechanism responsible for pathological changes in CVBF.

Photodynamic opening of blood-brain barrier

Photodynamic treatment (PDT) causes a significant increase in the permeability of the blood-brain barrier (BBB) in healthy mice. Using different doses of laser radiation (635 nm, 10-40 J/cm2) and photosensitizer (5-aminolevulinic acid - 5-ALA, 20 and 80 mg/kg, i.v.), we found that the optimal PDT for the reversible opening of the BBB is 15 J/cm2 and 5-ALA, 20 mg/kg, exhibiting brain tissues recovery 3 days after PDT. Further increases in the laser radiation or 5-ALA doses have no amplifying effect on the BBB permeability, but are associated with severe damage of brain tissues. These results can be an informative platform for further studies of new strategies in brain drug delivery and for better understanding of mechanisms underlying cerebrovascular effects of PDT-related fluorescence guided resection of brain tumor.

The Stress and Vascular Catastrophes in Newborn Rats: Mechanisms Preceding and Accompanying the Brain Hemorrhages

In this study, we analyzed the time-depended scenario of stress response cascade preceding and accompanying brain hemorrhages in newborn rats using an interdisciplinary approach based on: a morphological analysis of brain tissues, coherent-domain optical technologies for visualization of the cerebral blood flow, monitoring of the cerebral oxygenation and the deformability of red blood cells (RBCs). Using a model of stress-induced brain hemorrhages (sound stress, 120 dB, 370 Hz), we studied changes in neonatal brain 2, 4, 6, 8 h after stress (the pre-hemorrhage, latent period) and 24 h after stress (the post-hemorrhage period). We found that latent period of brain hemorrhages is accompanied by gradual pathological changes in systemic, metabolic, and cellular levels of stress. The incidence of brain hemorrhages is characterized by a progression of these changes and the irreversible cell death in the brain areas involved in higher mental functions. These processes are realized via a time-depended reduction of cerebral venous blood flow and oxygenation that was accompanied by an increase in RBCs deformability. The significant depletion of the molecular layer of the prefrontal cortex and the pyramidal neurons, which are crucial for associative learning and attention, is developed as a consequence of homeostasis imbalance. Thus, stress-induced processes preceding and accompanying brain hemorrhages in neonatal period contribute to serious injuries of the brain blood circulation, cerebral metabolic activity and structural elements of cognitive function. These results are an informative platform for further studies of mechanisms underlying stress-induced brain hemorrhages during the first days of life that will improve the future generations health.

Analysis of blood pressure dynamics in male and female rats using the continuous wavelet transform

We study gender-related particularities in cardiovascular responses to stress and nitric oxide (NO) deficiency in rats using HR, mean arterial pressure (MAP) and a proposed wavelet-based approach. Blood pressure dynamics is analyzed: (1) under control conditions, (2) during immobilization stress and recovery and (3) during nitric oxide blockade by N(G)-nitro-L-arginine-methyl ester (L-NAME). We show that cardiovascular sensitivity to stress and NO deficiency depends upon gender. Actually, in females the chronotropic effect of stress is more pronounced, while the pressor effect is weakened compared with males. We conclude that females demonstrate more favorable patterns of cardiovascular responses to stress and more effective NO control of cardiovascular activity than males.

Laser speckle contrast imaging of cerebral blood flow of newborn mice at optical clearing

In this work, we consider the use of optical clearing agents to improve imaging quality of the cerebral blood flow of newborn mice. Aqueous 60%-glycerol solution, aqueous 70%-OmnipaqueTM(300) solution and OmnipaqueTM (300) solution in water/DMSO(25%/5%) were selected as the optical clearing agents. Laser speckle contrast imaging (LSCI) was used for imaging of cerebral blood flow in newborn mice brain during topical optical clearing of tissuesin the area of the fontanelle. These results demonstrate the effectiveness of glycerol and Omnipaque solutions as optical clearing agents for investigation of cerebral blood flow in newborn mice without scalp removing and skull thinning.

Application of optical coherence tomography for in vivo monitoring of the meningeal lymphatic vessels during opening of blood–brain barrier: mechanisms of brain clearing

Abstract. The meningeal lymphatic vessels were discovered 2 years ago as the drainage system involved in the mechanisms underlying the clearance of waste products from the brain. The blood–brain barrier (BBB) is a gatekeeper that strongly controls the movement of different molecules from the blood into the brain. We know the scenarios during the opening of the BBB, but there is extremely limited information on how the brain clears the substances that cross the BBB. Here, using the model of sound-induced opening of the BBB, we clearly show how the brain clears dextran after it crosses the BBB via the meningeal lymphatic vessels. We first demonstrate successful application of optical coherence tomography (OCT) for imaging of the lymphatic vessels in the meninges after opening of the BBB, which might be a new useful strategy for noninvasive analysis of lymphatic drainage in daily clinical practice. Also, we give information about the depth and size of the meningeal lymphatic vessels in mice. These new fundamental data with the applied focus on the OCT shed light on the mechanisms of brain clearance and the role of lymphatic drainage in these processes that could serve as an informative platform for a development of therapy and diagnostics of diseases associated with injuries of the BBB such as stroke, brain trauma, glioma, depression, or Alzheimer disease.

Silent Vascular Catastrophes in the Brain in Term Newborns: Strategies for Optical Imaging

In this review, we describe the phenomenon of intra-cranial hemorrhage (ICH), which occurs spontaneously within the first 2-3 days of life in full-term newborns without any clinical symptoms, but with long-term neurological outcomes in many cases. We give the estimated frequency and possible mechanisms responsible for silent ICH with main focus on the cerebral blood flow (CBF) and oxygenation measurements using noninvasive optical methods. We overview the current multi-modal technologies that are widely used in clinics and experiments for the study of ICH during the first days of life: magnetic resonance imaging, ultrasonography, cerebral oximetry based on near infrared spectroscopy, as well as laser speckle imaging and diffuse correlation spectroscopy as a priority technology for experimental study of cerebral hemodynamics. We discuss the advantages and disadvantages of these methods. We identify key trends in experimental works and show areas for future research of ICH in term newborns. Future studies will help to improve our ability to optimize prognosis, diagnosis, and treatment of asymptomatic neonates with silent ICH.