Disturbed Blood Flow Research Articles

Nanotheranostics of Pre‐Stenotic Vessels By Target Touch‐On Signaling of Peptide Navigator

AbstractNavigation of nanoparticles to target sites of blood flow disturbance markedly upgrades the diagnostic paradigm in vascular medicine. The theranostic treatment of pre‐stenotic vessels can prevent the irreversible occlusion process effectively. Here, these nanotheranostic functions are established by displaying CDK9(cyclin‐dependent kinase 9)‐targeting peptide (P.) onto nanovesicles (NV) and liposomes using the navigation function and subsequent binding‐on signaling of P. as a game‐changer. When rabbit vessels are allografted with injecting contrast‐loaded P. liposomes, the case‐dependent stenotic degree after 2–6 weeks can be diagnosed accurately within 2–4 days via computed tomography imaging with cross‐validation in a mouse model of partial carotid ligation. Furthermore, the anti‐CDK9 signaling of P. NV is activated post‐targeting and effectively prevents vascular stenosis by suppressing inflammation and lipotoxicity in the vessels, serum, and/or liver. CDK9 targeting is confirmed using computer, in vitro, and in vivo models. This study demonstrates an unprecedented nanotheranostic function for future clinical applications.

Cerebroplacental ratio for prediction of adverse intrapartum and neonatal outcomes in a term uncomplicated pregnancy

BackgroundFetal hypoxia is one of the major causes of high perinatal morbidity and mortality rates. Doppler ultrasound tests such as cerebroplacental ratio (CPR) evaluation are commonly used to assess blood flow disturbances in placento-umbilical and feto-cerebral circulations. A low cerebroplacental ratio has been shown to be associated with an increased risk of stillbirth regardless of the gestation or fetal weight. We conducted this study to assess the fetal cerebroplacental ratio in prediction of adverse intrapartum and neonatal outcomes in a term, uncomplicated pregnancy to reduce fetal and neonatal morbidity and mortality.ResultsIt was found that neonates with CPR ≤1.1 had significantly higher frequencies of cesarean delivery (CS) for intrapartum fetal compromise compared to those with CPR >1.1 (p=0.043). Neonates with CPR ≤1.1 had significantly lower Apgar score at 1 min and 5 min than those with CPR >1.1 (p=0.004) and (p=0.003), respectively. Neonates with CPR ≤1.1 had significantly higher rates of NICU admission than those with CPR <1.1 (p=0.004).ConclusionThe cerebroplacental ratio shows the highest sensitivity in the prediction of fetal heart rate abnormalities and adverse neonatal outcome in uncomplicated pregnancies at term. The cerebroplacental ratio index is useful in clinical practice in antenatal monitoring of these women in order to select those at high risk of intra- and postpartum complications.

Особливості патогенетичних механізмів порушення регіонарного кровотоку при синдромі діабетичної стопи (огляд літератури)

Цукровий діабет (ЦД) у світі визнано однією з найбільш важливих неінфекційних хвороб, поширення якої набуло характеру пандемії. У переліку пізніх ускладнень ЦД лідирує синдром діабетичної стопи, призводячи до ранньої інвалідизації та визначаючи високий рівень летальності. За даними Міжнародної діабетичної федерації, від 25 до 47 % госпіталізацій хворих на ЦД пов’язано з гнійно-деструктивними ураженнями стоп унаслідок розвитку хронічної ішемії тканин. В огляді відображені основні патофізіологічні механізми виникнення порушення мікроциркуляції при синдромі діабетичної стопи, що призводять до виникнення критичної ішемії нижніх кінцівок.

Emerging Microfluidic Approaches for Platelet Mechanobiology and Interplay With Circulatory Systems.

Understanding how platelets can sense and respond to hemodynamic forces in disturbed blood flow and complexed vasculature is crucial to the development of more effective and safer antithrombotic therapeutics. By incorporating diverse structural and functional designs, microfluidic technologies have emerged to mimic microvascular anatomies and hemodynamic microenvironments, which open the floodgates for fascinating platelet mechanobiology investigations. The latest endothelialized microfluidics can even recapitulate the crosstalk between platelets and the circulatory system, including the vessel walls and plasma proteins such as von Willebrand factor. Hereby, we highlight these exciting microfluidic applications to platelet mechanobiology and platelet–circulatory system interplay as implicated in thrombosis. Last but not least, we discuss the need for microfluidic standardization and summarize the commercially available microfluidic platforms for researchers to obtain reproducible and consistent results in the field.

4D flow MRI hemodynamic biomarkers for cerebrovascular diseases.

Alterations in cerebral blood flow are common in several neurological diseases among the elderly including stroke, cerebral small vessel disease, vascular dementia, and Alzheimer's disease. 4D flow magnetic resonance imaging (MRI) is a relatively new technique to investigate cerebrovascular disease, and makes it possible to obtain time-resolved blood flow measurements of the entire cerebral arterial venous vasculature and can be used to derive a repertoire of hemodynamic biomarkers indicative of cerebrovascular health. The information that can be obtained from one single 4D flow MRI scan allows both the investigation of aberrant flow patterns at a focal location in the vasculature as well as estimations of brain-wide disturbances in blood flow. Such focal and global hemodynamic biomarkers show the potential of being sensitive to impending cerebrovascular disease and disease progression and can also become useful during planning and follow-up of interventions aiming to restore a normal cerebral circulation. Here, we describe 4D flow MRI approaches for analyzing the cerebral vasculature. We then survey key hemodynamic biomarkers that can be reliably assessed using the technique. Finally, we highlight cerebrovascular diseases where one or multiple hemodynamic biomarkers are of central interest.

Abstract 12602: Local NET Burden is Associated With Biomarkers of Thrombosis and Cardiac Damage in Left Atrial Cardiopathy

Atrial cardiopathy results in disturbed blood flow, providing an ideal thrombogenic environment. Neutrophils promote large vessel occlusions and microthrombosis via release of neutrophil extracellular traps (NETs) and thus lie at the interface of (sterile) inflammation, thrombosis, and fibrosis. We profiled NETs, cytokines, thrombotic factors, and cardiac damage markers in left atrial (LA) cardiopathies together with LA structural analysis. A total of 71 patients (median age 66, 46.5% male) undergoing catheterization procedures with atrial transseptal access at the Freiburg Heart Center were enrolled in this prospective study. Paired samples were collected from peripheral blood draws or from within the left atrium (LA). NET biomarkers, sP-selectin, PAD4, proinflammatory cytokines, thrombotic factors (D-dimers, VWF, ADAMTS13), and cardiac damage markers (TropT, proBNP, ICTP) were measured in plasma or serum as appropriate. Echocardiography was performed to assess LA structure and left ventricular function. Data were analyzed by the type of procedure (MitraClip (MC), left atrial appendage closure (LAAC), pulmonary vein ablation (PVA), or patent foramen ovale closure (PFOC), and by atrial fibrillation (Afib) status at time of procedure. NETs, but not other markers, were elevated in local LA samples as compared to peripheral blood. Conversely, most thrombotic, inflammatory, and cardiac damage markers, but not NETs, were elevated in MC or LAAC as compared to PFOC or PVA patients. Across all patients, NET biomarkers positively correlated with IL-6/TNF-α, VWF, and cardiac damage biomarkers. PAD4/NETs negatively correlated with ADAMTS13 activity, indicating that citrullination may contribute to the prothrombotic phenotype of these patients. Afib was not key to these observations. NETs are locally elevated within the left atrium, with elevation of thrombosis markers including VWF in patients with a diseased left atrium. These elevations partly correlate with structural changes of the left atrium and, therefore, may point toward a disturbed local blood flow. This may provide insight on mechanisms that drive detrimental left atrial remodeling as well as clues regarding thromboembolic risks that are rooted in LA changes rather than Afib.

Abstract 12466: Suppression of Inflammatory Activity in Vascular Endothelial Cells Reduces EndMT in Response to Oscillatory Shear Stress

Background: The incidence of pro-atherogenic lesions is greater in the branching regions of artery compared to non-branching regions. Disturbed blood flow in the atheroprone regions exerts oscillatory shear stress (OSS) to the vascular wall, particularly endothelial cells (ECs), and OSS is a risk factor for the development of pro-atherogenic lesions. This study aims to assess the impact of OSS on phenotypic transition in artery ECs. Methods and Results: Immunofluorescence staining and image quantification revealed that inflammatory activity and endothelial-to-mesenchymal transition (EndMT) are greater in mouse aortic arch than in the descending thoracic aorta. Inflammatory biomarker ICAM-1 is colocalized with the EndMT biomarker, vimentin and α-smooth muscle actin, in ECs of the aorta arch. As cells in aortic arch sustain OSS, we tested the hypothesis that OSS induces EndMT in artery ECs by a pro-inflammatory mechanism, using a three-dimensional microengineered human artery-on-a-chip system. Human coronary artery ECs were cultured on the surface of a collagen I-coated membrane and subjected to OSS. Single-cell RNA sequencing analysis revealed that inflammatory and EndMT-related genes were enriched in ECs after being exposed to OSS for 24 h. OSS-induced inflammatory response and EndMT were confirmed by immunoblotting and immunofluorescence staining. These changes were abolished by inhibiting the p38 MAPK-NF-κB signaling pathway. Moreover, anti-inflammatory cytokine, IL-37, suppressed the inflammatory response and prevented EndMT in ECs exposed to OSS. Importantly, expression of IL-37 in mice markedly reduced the level of inflammatory biomarker and the density of cells displaying EndMT biomarkers in the endothelium of aortic arch. Conclusion: OSS induces EndMT in artery ECs through a pro-inflammatory mechanism involving the p38 MAPK-NF-κB signaling pathway and contributes to the pro-atherogenic microenvironment in regions sustaining disturbed blood flow. Anti-inflammatory cytokine IL-37 may have the therapeutic potential for suppressing endothelial phenotypic transition caused by disturbed blood flow.

Abstract 9360: Increasing Mass Transfer Flux of Low-Density Lipoproteins-Cholesterol or Apolipoprotein B at the Endothelium of Atherogenic Sites as a Primary Target of Atherosclerotic Cardiovascular Disease

Major pathological evidences indicate that subendothelial low-density lipoproteins-cholesterol (LDL-c) accumulations at atherogenic sites cause atheroscleroticcardiovascular disease (ACD). As such, current guidelines focus on LDL-c as aprimary target of ACD. Accumulating data from clinical trials have suggested thata shift from LDL-c to apolipoprotein B (Apo B) may improve target accuracy. Inthis study, we assess the hypothesis that focuses on increasing mass transfer flux ofLDL-c or Apo B from the disturbed blood flow to the endothelium of atherogenicsites as a primary target of ACD. A set of multidisciplinary analysis models areused to describe dynamic behaviors of LDL-c or Apo B particles and create amultibiomarker analysis formula (MAF) for the assessment, prevention andtreatment of ACD. The models reveal that individuals’ atheroscleroticmultibiomarker such as elevated LDL-c levels or increasing Apo B, hypertension,rising heart rate and increasing atrial stiffness contribute to an increase in LDL-c orApo B mass transfer flux at the endothelium of atherogenic sites and the increasingLDL-c flux then directly triggers subendothelial LDL-c accumulations at the sites.Excitingly, we show that the individual ACD risks assessed by MAF correspondrobustly to the 2013 ACC/AHA Guideline. In conclusion, LDL-c or Apo B masstransfer flux at the endothelium of atherogenic sites is a primary target of ACD andthe flux combines the contributions of different biomarkers to ACD. MAFencompasses a multibiomarker approach that has been utilized to analyzescreening data from over 95,000 patients conducted by the Cleveland Heart Laband MDVIP, Inc., which highlights the potential to prevent ACD and savehundreds of millions of dollars in healthcare costs.

Features of heart failure with preserved ejection fraction (HFpEF) in diabetic patients with resistant hypertension

BACKGROUND: It is expected that a steady increase in the incidence of diabetes and resistant hypertension (RHTN), along with an increase in life expectancy, will lead to a noticeable increase in the proportion of patients with heart failure with preserved ejection fraction (HFpEF). At the same time, data on the frequency of HFpEF in a selective group of patients with RHTN in combination with diabetes are still lacking, and the pathophysiological and molecular mechanisms of its formation have not been yet studied sufficiently.AIM: To assess the features of the development HFpEF in diabetic and non-diabetic patients with RHTN, as well as to determine the factors associated with HFpEF.MATERIALS AND METHODS: In the study were included 36 patients with RHTN and type 2 diabetes mellitus (DM) (mean age 61.4 ± 6.4 years, 14 men) and 33 patients with RHTN without diabetes, matched by sex, age and level of systolic blood pressure (BP). All patients underwent baseline office and 24-hour BP measurement, echocardiography with assess diastolic function, lab tests (basal glycemia, HbA1c, creatinine, aldosterone, TNF-alpha, hsCRP, brain naturetic peptide, metalloproteinases of types 2, 9 (MMP-2, MMP-9) and tissue inhibitor of MMP type 1 (TIMP-1)). HFpEF was diagnosed according to the 2019 AHA/ESC guidelines.RESULTS: The frequency of HFpEF was significantly higher in patients with RHTN with DM than those without DM (89% and 70%, respectively, p=0.045). This difference was due to a higher frequency of such major functional criterion of HFpEF as E/e’≥15 (p=0.042), as well as a tendency towards a higher frequency of an increase in left atrial volumes (p=0.081) and an increase in BNP (p=0.110). Despite the comparable frequency of diastolic dysfunction in patients with and without diabetes (100% and 97%, respectively), disturbance of the transmitral blood flow in patients with DM were more pronounced than in those without diabetes. Deterioration of transmitral blood flow and pseudo-normalization of diastolic function in diabetic patients with RHTN have relationship not only with signs of carbohydrate metabolism disturbance, but also with level of pulse blood pressure, TNF-alfa, TIMP-1 and TIMP-1 / MMP-2 ratio, which, along with the incidence of atherosclerosis, were higher in patients with DM than in those without diabetes.CONCLUSIONS: Thus, HFpEF occurs in the majority of diabetic patients with RHTN. The frequency of HFpEF in patients with DN is significantly higher than in patients without it, which is associated with more pronounced impairments of diastolic function. The progressive development of diastolic dysfunction in patients with diabetes mellitus is associated not only with metabolic disorders, but also with increased activity of chronic subclinical inflammation, profibrotic state and high severity of vascular changes.

Revisited Hyperoxia Pathophysiology in the Perioperative Setting: A Narrative Review.

The widespread use of high-dose oxygen, to avoid perioperative hypoxemia along with WHO-recommended intraoperative hyperoxia to reduce surgical site infections, is an established clinical practice. However, growing pathophysiological evidence has demonstrated that hyperoxia exerts deleterious effects on many organs, mainly mediated by reactive oxygen species. The purpose of this narrative review was to present the pathophysiology of perioperative hyperoxia on surgical wound healing, on systemic macro and microcirculation, on the lungs, heart, brain, kidneys, gut, coagulation, and infections. We reported here that a high systemic oxygen supply could induce oxidative stress with inflammation, vasoconstriction, impaired microcirculation, activation of hemostasis, acute and chronic lung injury, coronary blood flow disturbances, cerebral ischemia, surgical anastomosis impairment, gut dysbiosis, and altered antibiotics susceptibility. Clinical studies have provided rather conflicting results on the definitions and outcomes of hyperoxic patients, often not speculating on the biological basis of their results, while this review highlighted what happens when supranormal PaO2 values are reached in the surgical setting. Based on the assumptions analyzed in this study, we may suggest that the maintenance of PaO2 within physiological ranges, avoiding unnecessary oxygen administration, may be the basis for good clinical practice.

Thromboinflammatory profiling in atrial cardiopathy

Abstract Background Thrombotic cardioemboli originate largely from a diseased left atrium. Disturbed blood flow in atrial cardiopathy is proposed to create a thrombogenic environment and enhance clot formation, even independently of atrial fibrillation. The formation of neutrophil extracellular traps (NETs) has been identified as a crucial mechanism in thrombotic diseases, including cardioembolic thrombus formation. Moreover, emerging evidence suggests that von Willebrand factor (VWF) plays a vital role in NET-mediated pathologies, serving as a bridge between activated endothelium, platelets, and leukocytes. VWF and NETs have also both been shown to be involved in fibrotic remodeling of the heart. Purpose of the study The aim of this study is to identify thrombotic markers that correlate with structural abnormalities of the heart. This may provide insight into pathomechanisms that drive detrimental left atrial remodeling as well as clues regarding thromboembolic risks that are rooted in atrial cardiopathy Methods Paired blood samples were collected from patients undergoing catheterization with atrial transseptal puncture. We compared blood sampled locally from the left atrium to peripheral blood samples collected prior to the catheterization procedure from the same patients, analyzing them for a wide range of thrombotic markers, including: NETs biomarkers, VWF, ADAMTS13, D-dimers, fibrinogen, as well as indicators of fibrosis like BNP and C-terminal telopeptide of type I collagen (ICTP). In parallel, echocardiographic measurements including left atrium size and ejection fraction of the left ventricle were recorded in order to perform correlation analyses with the biochemical parameters in blood. Results A total of 71 patients (age 64,1±14, males 46,5%) were included in the study. Myeloperoxidase (MPO)-DNA complexes and citrullinated histone H3 (H3Cit) levels were significantly elevated in samples drawn from the left atrium compared to peripheral samples, and MPO-DNA levels in the left atrium correlated with left atrial size (Figure 1). H3Cit and PAD4 levels correlated with VWF activity and antigen, in central and peripheral samples. Reduced peripheral ADAMTS13 activity levels correlated with higher values throughout all NET markers, and PAD4, which has been shown to render ADAMTS13 inactive (Figure 2). NETs and PAD4 also correlated with BNP and ICTP, providing potential novel biomarkers for fibrosis development. Conclusion Blood drawn from the left atrium shows significantly higher levels of specific (not all) thrombotic markers as compared to blood drawn from peripheral veins, and a relationship between NETs and dysregulation of the VWF-ADAMTS13 axis. These elevations partly correlate with structural changes of the left atrium and, therefore, may point toward a disturbed local blood flow. This may help identify patients at risk for cardioembolic events, even independently of atrial fibrillation. Funding Acknowledgement Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Research Area Network - Cardio Vascular Disease (ERA-CVD)Fonds Wetenschapplijk Onderzoek Vlaanderen (FWO)

Isolation of Endothelial Cells from the Lumen of Mouse Carotid Arteries for Single-cell Multi-omics Experiments.

Atherosclerosis is an inflammatory disease of the arterial regions exposed to disturbed blood flow (d-flow). D-flow regulates the expression of genes in the endothelium at the transcriptomic and epigenomic levels, resulting in proatherogenic responses. Recently, single-cell RNA sequencing (scRNAseq) and single-cell Assay for Transposase Accessible Chromatin sequencing (scATACseq) studies were performed to determine the transcriptomic and chromatin accessibility changes at a single-cell resolution using the mouse partial carotid ligation (PCL) model. As endothelial cells (ECs) represent a minor fraction of the total cell populations in the artery wall, a luminal digestion method was used to obtain EC-enriched single-cell preparations. For this study, mice were subjected to PCL surgery to induce d-flow in the left carotid artery (LCA) while using the right carotid artery (RCA) as a control. The carotid arteries were dissected out two days or two weeks post PCL surgery. The lumen of each carotid was subjected to collagenase digestion, and endothelial-enriched single cells or single nuclei were obtained. These single-cell and single-nuclei preparations were subsequently barcoded using a 10x Genomics microfluidic setup. The barcoded single-cells and single-nuclei were then utilized for RNA preparation, library generation, and sequencing on a high-throughput DNA sequencer. Post bioinformatics processing, the scRNAseq and scATACseq datasets identified various cell types from the luminal digestion, primarily consisting of ECs. Smooth muscle cells, fibroblasts, and immune cells were also present. This EC-enrichment method aided in understanding the effect of blood flow on the endothelium, which could have been difficult with the total artery digestion method. The EC-enriched single-cell preparation method can be used to perform single-cell omics studies in EC-knockouts and transgenic mice where the effect of blood flow on these genes has not been studied. Importantly, this technique can be adapted to isolate EC-enriched single cells from human artery explants to perform similar mechanistic studies.

Study on viscosity induced contrast in ultrasound color flow imaging of carotid atherosclerosis

Efficient imaging of blood flow disturbances resulted from carotid atherosclerosis plays a vital role clinically to predict brain stroke risk. Carotid atherosclerosis and its development is closely linked with raised blood viscosity. Therefore, study of viscosity changing hemodynamic effect has importance and it might be useful for improved examination of carotid atherosclerosis incorporating the viscosity induced contrast in conventional ultrasound imaging. This work considered the design of realistic models of atherosclerotic carotid artery of different stages and solved to compute the hemodisturbances using computational fluid dynamics (CFD) by finite element method (FEM) to investigate viscosity changes effect. Ultrasound color flow image of velocities of blood have been constructed using phase shift information estimated with autocorrelation of Hilbert transformed simulated backscattered radiofrequency (RF) signals from moving blood particles. The simulated ultrasound images have been compared with CFD simulation images and identified a good match between them. The atherosclerosis stages of the models have been investigated from the estimated velocity data. It has been observed that the blood velocities increase noticeably in carotid atherosclerotic growths and velocity distribution changes with viscosity variations. It is also found importantly that the viscosity induced contrast associated to atherosclerosis is detectable in ultrasound color flow imaging. The findings of this work might be useful for better investigation of carotid atherosclerosis as well as prediction of its progression to reduce the stroke risk.

Trophoblastic volumetric bloodflow evaluation in women with trombophylia on 7–10th week of pregnancy

Spontaneous abortion occur in 15–20% of all detected pregnancies, and 45–70% of all spontaneous miscarriages take place up to 10 weeks. Thrombophilia is one of the main causes of miscarriage and its effect on the course of pregnancy remains poorly understood. In this regard, there is great interest in the problem of diagnosing thrombophilia and further prevention of thrombosis in obstetric practice.Purpose. Compare the parameters of volumetric blood flow in the chorion with different types of trophoblastic blood flow disturbance in pregnant women with thrombophilia in the first trimester for their quantitative verification.Materials and methods. The study included 129 pregnant women at the 7th to 10th week of gestation with a diagnosis of thrombophilia and a control group - pregnant women with a normal course of this pregnancy and a successful outcome of past pregnancies.The patients were divided into III clinical groups:I. Pregnant women with a normal course of this pregnancy and a successful outcome of past pregnancies (n = 33) – comparison group;II. Pregnant women with thrombophilia, in whom current pregnancy occurred on anticoagulant therapy (n = 28);III. Pregnant women with thrombophilia without taking anticoagulant drugs before registration of pregnancy (n = 68).Ultrasound was performed on Voluson E8 and S8 machines. All pregnant women underwent transabdominal and transvaginal ultrasound in B-mode, in color-doppler (CD) mode, and volumetric reconstruction of chorion was performed using the VOCAL program. In B-mode, the state of the embryo and extraembryonic structures were examined. The identification of trophoblastic vessels was carried out using the CD – region of interest was placed in the trophoblast area detected in B-mode, size and shape was adapted for a particular section.Using three-dimensional echography, the chorion volume was performed. The degree of blood supply in chorionic volume were calculated; vascularization index (VI), flow index (FI) and perfusion index (VFI) were displayed on a histogram with quantitative indicators.ResultsIn the CD mode, the types of trophoblastic blood flow were identified in the chorionic vessels:1) continuous type – blood flow loci are continuously identified over the entire area of the basal surface of the trophoblast;2) intermittent type – blood flow loci are unevenly identified along the basal surface of the trophoblast;3) single loci – single blood flow loci are identified along the basal surface of the trophoblast;4) lack of blood flow – blood flow loci are not determined along the basal surface of the trophoblast.Assessment of the quantitative parameters of the volumetric blood flow was used in the corresponding type of blood flow to verify the classification of types of trophoblastic blood flow in the CD mode.Statistical analysis of the volumetric blood flow indices using three-dimensional chorionic reconstruction showed a high degree of reliability (p &lt; 0.0001) in the VI and VFI values between all corresponding types of blood flow.Discussion. High incidence of spontaneous abortion and the significant role of thrombophilia in this pathology leads to necessity of expanding the research in this area. There has been a lot of research done over the past decade. The quantitative threshold values of the resistance indices in the chorionic vessels were calculated using pulse-wave Doppler, the chorionic blood supply indices were determined using volumetric reconstruction for various pathologies.However, we have not previously met the classification of chorionic blood supply in the CD mode. This technique is very simple to perform and is a highly informative for predicting miscarriage in the first trimester. For the first time, we proposed the above classification of trophoblastic blood flow; for the first time, we proved the reliability of the classification developed by us using the method of volumetric reconstruction of the chorion with an assessment of trophoblastic blood flow, which allows us to recommend its use when writing a protocol for ultrasound of the fetus in the first trimester in pregnant women with thrombophilia.Conclusion. The classification of trophoblastic blood flow was verified by three-dimensional echography with quantitative indicators of the volumetric blood flow of the trophoblast vessels, with a high degree of reliability (p &lt; 0.001) which to indicates the reliability of this classification. It can be recommended to conduct a multicenter study for investigating fetal ultrasound with the additional use of the CD in the first trimester in pregnant women with a diagnosis of thrombophilia.

Abstract 127: Aspirin Resistant Biomechanical Platelet Activation In Fibromuscular Dysplasia: Novel Mechanisms Of Platelet Activation &amp; Stroke

Background: Fibromuscular dysplasia (FMD) is a non-atherosclerotic, non-inflammatory disorder characterized by abnormal arterial morphology and turbulent blood flow. In regions of disturbed blood flow, circulating platelets may become activated. Cerebrovascular FMD is common and increases the risk of stroke in affected patients. Most FMD patients are treated with antiplatelet agents, however these medications target biochemical and not biomechanical pathways of platelet activation. Methods: Thrombotic outcomes were determined by multivariate regression analysis of 105,887 patients with FMD. Platelets were isolated from humans and activation was assessed by FACS and aggregometry following exposure to ex vivo steady laminar and disturbed flow (SF and DF, respectively) conditions using a cone and flow system. Single-cell phenotyping of control and FMD platelets with transcriptomics (RNAseq) and proteomics were performed. Cellular bioenergetics were evaluated by real-time metabolic analyses. Results: In patients with FMD, long-term aspirin therapy is paradoxically an independent risk factor for acute ischemic stroke (OR 1.64, 95% CI 1.29-1.94) but seems to protect against acute hemorrhagic stroke (OR 0.47, 95% CI 0.24-0.89). FMD platelet activation and aggregation were hyporeactive to biochemical agonists, but hyperreactive to ex vivo mechanical, disturbed blood flow conditions compared to age/gender-matched controls. FMD platelet activation in DF was attenuated by the mechanosensitive channel inhibitor, GsMTx4. Principle component analyses of RNAseq and proteomics identified markedly different platelet phenotypes with key differences in mitochondrial function/fission. Platelet imaging and metabolic analyses revealed abnormal mitochondrial architecture and respiration in FMD. Conclusions: FMD platelets display a divergent, dysregulated platelet phenotype with attenuated biochemical but augmented biomechanical activation. DF exposure of platelets in irregularly shaped arteries in FMD may alter the platelet phenotype and metabolic function. Therapeutics targeting biomechanical pathways of platelet activation may provide a superior mechanism for stroke prevention without unintended bleeding consequences.

Visualization of Carotid Doppler in Patients with Ischemic Stroke at Dr. Hasan Sadikin General Hospital Bandung Year 2016-2019

Background: Various pathological changes in both the intra and extracranial arteries that supply the brain can cause disturbance of cerebral blood flow and perfusion leading to cerebral dysfunction. Doppler ultrasound is able to assess these changes. This study was performed to evaluate the anatomical and physiological changes found in the carotid arteries of patients with ischemic stroke using Doppler ultrasound. Met h ods: The cross-sectional descriptive study design with total sampling method was conducted on the medical records of ischemic stroke patients who had carotid Doppler ultrasound at the Department of Cardiology and Vascular Medicine Dr. Hasan Sadikin General Hospital Bandung from 2016 to 2019. Demographic data, such as stroke diagnoses and plaque characteristics recorded in the Doppler reports were collected. Result s : There were 38 data sets collected. The distribution and characteristics of atherosclerotic plaques were similar between the two carotid systems, with the same percentage of plaque being found in the right (31.6%) and left (36.8%) carotid system. The most common type of plaque found was type III and was located in the common carotid artery. Thrombus was absent in all patients. Intimal media thickening was found in 13.2% right system and 15.8% left system. Stenosis was present in 34.2% of patients, and most had &lt;50% stenosis. Peak systolic velocity increased ( &gt; 125 cm/s) in 5.3% of the right system and 7.9% of the left system of the internal carotid artery. Conclusion s : Most of the atherothrombotic and thromboembolic type of ischemic stroke patients in this study have normal carotid Doppler ultrasound features. Further study on the presence of plaque in ischemic stroke patients in Indonesia is needed.

Abstract MP45: The Roles Of Telomeric Repeat Binding Factor 2-interacting Protein (TERF2IP) K240 Sumoylation In Endothelial Cells On Atherogenesis

Background and Objectives: It is not yet clear how the pro-atherogenic signaling events in endothelial cells (ECs) such as those that lead to EC senescence, apoptosis and activation are interconnected and promote atherosclerotic plaque formation in the area exposed to disturbed blood flow (d-flow). TERF2IP, a member of the shelterin complex of the telomere, regulates all three pathological events. We investigated the role of TERF2IP K240 SUMOylation in the process of d-flow-induced atherosclerotic plaque formation. Methods and Results: We found that d-flow increased TERF2IP K240 SUMOylation in ECs and that it was suppressed by a p90RSK specific inhibitor, FMK-MEA. This SUMOylation was independent of TERF2IP S205 phosphorylation. The d-flow-induced senescence, DNA damage, and apoptosis were inhibited in ECs with TERF2IP depletion or point-mutated phosphorylation (S205A) and SUMOylation (K240R) sites. NF-κB activation induced by d-flow or overexpression of p90RSK was also significantly inhibited in ECs overexpressing the TERF2IP S205A phosphorylation mutant. However, cells overexpressing the TERF2IP K240R SUMOylation mutant showed no effect on the d-flow or p90RSK-medaited NF-κB activation. To determine the biological function of TERF2IP K240 SUMOylation, we generated TERF2IP K240R knock-in (KI) mice and examined d-flow-induced atherosclerotic plaque formation using partial left carotid ligation model mice fed a high-fat diet after AAV8-PCSK9 injection. We found no differences in body weights and cholesterol levels between TERF2IP K240R KI and wild type control (WT) mice, but plaque formation was significantly inhibited in the KI mice compared to WT animals (Oil Red O positive area (%): 19.0 +/- 12.5 (KI mice, n=8) vs 61.2 +/- 24.3 (WT mice, n=7), p = 0.0008). Bone marrow from WT mice were transplanted into KI and WT mice, which were then injected with AAV8-PCSK9 virus and fed a high-fat diet for 16 weeks, but we still found that plaque formation was inhibited in the KI mice. Conclusion: TERF2IP SUMOylation plays a role in in EC senescence but not in activation. The significant inhibition of plaque formation in the TERF2IP K240R KI mice is due to downregulation of TERF2IP SUMOylation in ECs not in myeloid cells.

Контроль стану кровообігу нижніх кінцівок у хворих на цукровий діабет

Дослідження проблеми ураження нижніх кінцівок у хворих на цукровий діабет (ЦД) набуває все більшого значення у зв’язку зі значним поширенням кількості хворих. Важливим є питання ранньої діагностики діабетичної ангіопатії нижніх кінцівок. У проявах хвороби Рейно й ураження нижніх кінцівок у хворих на ЦД є багато спільного. При хворобі Рейно, як правило, спостерігається спазм дрібних артерій дистальних відділів кінцівок. У хворих на ЦД відбувається порушення магістрального кровотоку в артеріях нижніх кінцівок різного ступеня тяжкості, що також призводить до спазму дрібних артерій дистальних відділів. Впровадження термографічної діагностики дозволяє диференціювати хворобу Рейно від проявів ускладнень ЦД.

Increased cerebral endothelium-dependent vasodilation in rats in the postcardiac arrest period.

Cardiovascular lability is common after cardiac arrest. We investigated whether altered endothelial function is present in cerebral and mesenteric arteries 2 and 4 h after resuscitation. Male Sprague-Dawley rats were anesthetized, intubated, ventilated, and intravascularly catheterized whereupon rats were randomized into four groups. Following 7 min of asphyxial cardiac arrest and subsequent resuscitation, cardiac arrest and sham rats were observed for either 2 or 4 h. Neuron-specific enolase levels were measured in blood samples. Middle cerebral artery segments and small mesenteric arteries were isolated and examined in microvascular myographs. qPCR and immunofluorescence analysis were performed on cerebral arteries. In cerebral arteries, bradykinin-induced vasodilation was inhibited in the presence of either calcium-activated K+ channel blockers (UCL1684 and senicapoc) or the nitric oxide (NO) synthase inhibitor, Nω-nitro-L-arginine methyl ester hydrochloride (l-NAME), whereas the combination abolished bradykinin-induced vasodilation across groups. Neuron-specific enolase levels were significantly increased in cardiac arrest rats. Cerebral vasodilation was comparable between the 2-h groups, but markedly enhanced in response to bradykinin, NS309 (an opener of small and intermediate calcium-activated K+ channels), and sodium nitroprusside 4 h after cardiac arrest. Endothelial NO synthase and guanylyl cyclase subunit α-1 mRNA expression was unaltered after 2 h, but significantly decreased 4 h after resuscitation. In mesenteric arteries, the endothelium-dependent vasodilation was comparable between corresponding groups at both 2 and 4 h. Our findings show enhanced cerebral endothelium-dependent vasodilation 4 h after cardiac arrest mediated by potentiated endothelial-derived hyperpolarization and NO pathways. Altered cerebral endothelium-dependent vasodilation may contribute to disturbed cerebral perfusion after cardiac arrest.NEW & NOTEWORTHY This is the first study, to our knowledge, to demonstrate enhanced endothelium-dependent vasodilation in middle cerebral arteries in a cardiac arrest rat model. The increased endothelium-dependent vasodilation was a result of potentiated endothelium-derived hyperpolarization and endothelial nitric oxide pathways. Immunofluorescence microscopy confirmed the presence of relevant receptors and eNOS in cerebral arteries, whereas qPCR showed altered expression of genes related to guanylyl cyclase and eNOS. Altered endothelium-dependent vasoregulation may contribute to disturbed cerebral blood flow in the postcardiac arrest period.

АНГИОПУЛЬМОНОГРАФИЯ С НИТРОГЛИЦЕРИНОВЫМ ТЕСТОМ В ДИАГНОСТИКЕ ОСТРЫХ ИНФЕКЦИОННЫХ ДЕСТРУКЦИЙ ЛЕГКИХ

Objective. To develop a method for additional and differential diagnosis of acute infectious lung destruction (AILD) based on angiopulmonography with the nitroglycerin test. Methods. Angiopulmonography with the nitroglycerin test was used in 10 patients with suppurative diseasesof thelung and pleura for additional and differential diagnosis of AILD The method was used in such situations when chest computed tomography did not allow to determine unambiguously the presence and / or prevalence of necrosis of the lung parenchyma. Results. In 3 patients with the lung abscess, a clear restriction of the decay cavity was registered with the preservation of the main blood flow and weakening of the parenchymal phase of the blood circulation along the periphery of the destructive area. During the nitroglycerin test performance there was no change in the filling of the microvascular bed with contrast along the periphery of the decay cavity, which made it possible to determine the presence of parietal sequesters. According to the results of the study, the lung gangrene was diagnosed in 6 patients. At the same time, two variants of circulatory disorders were noted: the first - with preservation of the blood flow through the main vessels and with the absence of a parenchymal phase in the lesion focus, the second - with the violation of the main blood flow. In the affected area no change in blood flow was observed after the nitroglycerin test performance. Similar results of the study indicated the development of necrosis of the pulmonary parenchyma, which was subsequently confirmed during the operations performed. In the site of inflammatory infiltration of the pulmonary parenchyma with preserved main blood flow, the depletion of the parenchymal phase of blood circulation was determined, but after the nitroglycerin test, a pronounced enrichment of the vascular architecture to the parenchymal phase in the pneumonia affecting part of the lung was noted. Conclusion. It has been established that AILD is characterized by irreversible changes in the vascular bed of the lung parenchyma in the lesion focus. Angiopulmonography with the nitroglycerin test is considered to be an additional highly informative method improving the early and differential diagnosis of AILD in difficult clinical situations. What this paper adds It has been found out that during angiopulmonography the areas of pulmonary necrosis are characterized by the absence of a vascular pattern with or without disturbance of the blood flow through the segmental arteries. At the same time, in contrast to the foci of pneumonia, the nitroglycerin test is not accompanied by an evaluation of the filling of the pulmonary vascular bed in the affected area, i.e. blood supply disorders are irreversible. Thus, based on an assessment of the nature and reversibility of the blood flow disturbances in the affected lung, it is possible to carry out differential diagnosis of the early stages of acute infectious lung destruction (AILD) and pneumonia.