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BACKGROUND AND PURPOSE: Intraplaque hemorrhage contributes to lipid core enlargement and plaque progression, leading to plaque destabilization and stroke. The mechanisms that contribute to the development of intraplaque hemorrhage are not completely understood. A higher incidence of intraplaque hemorrhage and thin/ruptured fibrous cap (upstream of the maximum stenosis in patients with severe [?70%] carotid stenosis) has been reported. We aimed to noninvasively study the distribution of intraplaque hemorrhage and a thin/ruptured fibrous cap in patients with mild-to-moderate carotid stenosis. MATERIALS AND METHODS: Eighty-eight symptomatic patients with stroke (

The endothelialization of gas exchange membranes can increase the hemocompatibility of extracorporeal membrane oxygenators and thus become a long-term lung replacement option. Cell seeding on large or uneven surfaces of oxygenator membranes is challenging, with cell aerosolization being a possible solution. In this study, we evaluated the endothelial cell aerosolization for biohybrid lung application. A Vivostat® system was used for the aerosolization of human umbilical vein endothelial cells with non-sprayed cells serving as a control. The general suitability was evaluated using various flow velocities, substrate distances and cell concentrations. Cells were analyzed for survival, apoptosis and necrosis levels. In addition, aerosolized and non-sprayed cells were cultured either static or under flow conditions in a dynamic microfluidic model. Evaluation included immunocytochemistry and gene expression via quantitative PCR. Cell survival for all tested parameters was higher than 90%. No increase in apoptosis and necrosis levels was seen 24 h after aerosolization. Spraying did not influence the ability of the endothelial cells to form a confluent cell layer and withstand shear stresses in a dynamic microfluidic model. Immunocytochemistry revealed typical expression of CD31 and von Willebrand factor with cobble-stone cell morphology. No change in shear stress-induced factors after aerosolization was reported by quantitative PCR analysis. With this study, we have shown the feasibility of endothelial cell aerosolization with no significant changes in cell behavior. Thus, this technique could be used for efficient the endothelialization of gas exchange membranes in biohybrid lung applications.

Flow features of a meandering river can vary seasonally and widely in the boreal zone, as seasonal variation in discharges affect the properties of meander bends and the spatial location of flow magnitudes. However, a better understanding of the impacts of consecutive high-discharge events in the boreal zone requires further analyses. Thus, the impacts of the fluvial processes and their magnitudes on different types of meander bends (e.g., sinuosity, symmetricity) were analysed throughout an open-channel flow period, using accurate flow measurements and hydrodynamic (two-dimensional) modelling. In particular, the research compared the impacts of different discharge events in magnitude and length of river evolution, and whether there is a difference in the erosion forces of the rising and falling phases of discharge hydrographs, and which of the phases has the greatest relevance to erosion forces. A case study was carried out in a middle boreal zone meandering river (Koitajoki in eastern Finland) during the 2020 open water season from May to October. Five different types of meander bends were considered in the study area. Flow magnitudes and shear forces were at their strongest during the spring flood peak, which is normally considered as the highest flow event during the hydrological year. Exceptionally, the largest summer discharge event reached a level that was close to the spring floods' maximum erosional capabilities. Bed shear stress values indicated the falling phases of the flood hydrographs are more important to river evolution than during the ascent stage, as the long duration of the flood downstage contributes more strongly to meander bend erosion and development. The smaller the curvature and larger the sinuosity of the bends, the more variation in erosional forces occurs between discharge stages. The study provides new insights into seasonal fluvial processes in boreal river meander bends as well as reinforces results from earlier river studies.

Human induced pluripotent stem cell-derived kidney organoids have potential for disease modeling and to be developed into clinically transplantable auxiliary tissue. However, they lack a functional vasculature, and the sparse endogenous endothelial cells (ECs) are lost upon prolonged culture in vitro, limiting maturation and applicability. Here, we use intracoelomic transplantation in chicken embryos followed by single-cell RNA sequencing and advanced imaging platforms to induce and study vasculogenesis in kidney organoids. We show expansion of human organoid-derived ECs that reorganize into perfused capillaries and form a chimeric vascular network with host-derived blood vessels. Ligand-receptor analysis infers extensive potential interactions of human ECs with perivascular cells upon transplantation, enabling vessel wall stabilization. Perfused glomeruli display maturation and morphogenesis to capillary loop stage. Our findings demonstrate the beneficial effect of vascularization on not only epithelial cell types, but also the mesenchymal compartment, inducing the expansion of ´on target´ perivascular stromal cells, which in turn are required for further maturation and stabilization of the neo-vasculature. The here described vasculogenic capacity of kidney organoids will have to be deployed to achieve meaningful glomerular maturation and kidney morphogenesis in vitro. ispartof: NPJ REGENERATIVE MEDICINE vol:7 issue:1 ispartof: location:United States status: published

Objective Arteriovenous grafts (AVGs) are the second best option for haemodialysis access when native arteriovenous fistulae placement is not possible, because they have a lower patency owing to neointimal hyperplasia at the venous anastomosis. This review aimed to evaluate the effect of geometric graft modification to the graft–vein interface on AVG patency. Data sources The MEDLINE and Embase (OvidSP) databases were systematically searched for relevant studies analysing the effect of geometrically modified AVGs on graft patency and stenosis formation (last search July 2019). Review methods Data regarding AVG type, patency, and graft outlet stenosis was extracted for further evaluation. Data were pooled in a random effects model to estimate the relative risk of graft occlusion within one year. Follow up, number of patients, and relevant patient characteristics were extracted for the quality assessment of the included studies using Newcastle-Ottawa Scale and Cochrane Risk of Bias Tool. The quality of the evidence was determined according to the Grading of Recommendations, Assessment, Development and Evaluations (GRADE) system. Results Search strategies produced 2772 hits, of which eight articles met predetermined inclusion criteria. Overall, the included articles had low to moderate risk of bias. In total, 414 expanded polytetrafluoroethylene AVGs (232 geometrically modified and 182 standard) were analysed, comprising two modified AVG types: a prosthetic cuff design (Venaflo®) and grafts with a Tyrell vein patch. Overall, modified grafts did not show a statistically significantly higher one year primary (relative risk [RR] 0.86, 95% confidence interval [CI] 95% 0.64–1.16; GRADE: “low to very low”) or secondary patency (RR 0.57, 95% CI 0.32–1.02; GRADE: “low to very low”) when compared with standard AVGs. Analysis of prosthetic cuffed grafts (112 patients) separately demonstrated a statistically significantly higher one year primary (RR 0.75, 95% CI 0.61–0.91) and one year secondary patency (RR 0.47, 95% CI 0.30–0.75) compared with standard grafts (92 patients). The results on stenosis formation were inconclusive and inadmissible to quantitative analyses. Conclusion The meta-analysis showed that a prosthetic cuff design significantly improves AVG patency, while a venous cuff does not. Although the heterogeneity and low number of available studies limit the strength of the results, this review shows the potential of grafts with geometric modification to the graft–vein anastomosis and should stimulate further clinical and fundamental research on improving graft geometry to improve graft patency.

Purpose Transport of water and solutes across vascular endothelium is important in normal physiology and critical in the development of various diseases, including atherosclerosis. However, there is debate about the routes for such transport. We recently showed that an albumin-sized tracer crossed endothelium at bicellular and tricellular junctions, a tracer having the size of high density lipoprotein crossed only through tricellular junctions, and a tracer with the size of low density lipoprotein was unable to cross by either route and instead traversed the cells themselves. Here we review previous work on the structure and function of tricellular junctions. We then describe a study in which we assessed the role of such junctions in the transport of an albumin-sized tracer. Methods We examined normal endothelial monolayers, the effect of agonists that modify their permeability, and the influence of different patterns of shear stress. Results Under normal conditions, approximately 85% of transendothelial transport occurred through tricellular junctions. This fraction was unchanged when permeability was reduced by sphingosine-1-phosphate or increased by thrombin, and also did not differ between endothelium exposed to multidirectional as opposed to uniaxial shear stress despite a > 50% difference in permeability. Conclusion These data show that tricellular junctions dominate normal transport of this tracer and largely determine influences of agonists and shear. The effects were attributable to changes in both the number and conductivity of the junctions. Further investigation of these structures will lead to increased understanding of endothelial barrier function and may suggest new therapeutic strategies in disease.

Regardless of beneficial associated with internal waste dump (IWD) method, practices of this method within boundaries of pit-slope have some serious problems on stability issues due to this area is zone of potential failure. This zone is known as dynamic reactive zone which is easy to deform by external force, and inherent dangers of failure posing a threat to slope. Therefore, it is paramount to study the induced shear stress behavior in this zone particularly when IWD method is adopted within this zone. In this paper, a numerical study for investigating IWD-induced shear stress behavior has been carried out using Finite Element Method (FEM) with Strength Reduction approach. Different scenarios as per pit-slope depths, IWD heights and buffer zone lengths have been accounted and simulated using PHASE 2 to understand changes in induced shear stress imposed on the pit-slope. It is found that shear stress imposed on pit slope seems change dramatically with increasing IWD height for case of buffer zone length is less than 100-m-long.

Organs-on-chips (OoCs) are systems containing engineered or natural miniature tissues grown inside microfluidic chips. To better mimic human physiology, the chips are designed to control cell microenvironments and maintain tissue-specific functions. Combining advances in tissue engineering and microfabrication, OoCs have gained interest as a next-generation experimental platform to investigate human pathophysiology and the effect of therapeutics in the body. There are as many examples of OoCs as there are applications, making it difficult for new researchers to understand what makes one OoC more suited to an application than another. This Primer is intended to give an introduction to the aspects of OoC that need to be considered when developing an application-specific OoC. The Primer covers guiding principles and considerations to design, fabricate and operate an OoC, as well as subsequent assaying techniques to extract biological information from OoC devices. Alongside this is a discussion of current and future applications of OoC technology, to inform design and operational decisions during the implementation of OoC systems.

Cerebral hemodynamics play an important role in the development of cerebrovascular diseases. In this work, we propose a numerical framework for modeling patient-specific cerebral blood flow, using commonly available clinical datasets. Our hemodynamic model was developed using Simscape Fluids library in Simulink, based on a block diagram language. Medical imaging data obtained from computerized tomography angiography (CTA) in 59 patients with aneurysmal subarachnoid hemorrhage was used to extract arterial geometry parameters. Flow information obtained from transcranial Doppler (TCD) measurement was employed to calibrate input parameters of the hemodynamic model. The results show that the proposed numerical model can reproduce blood flow in the circle of Willis (CoW) per patient per measurement set. The resistance at the distal end of each terminal branch was the predominant parameter for the flow distribution in the CoW. The proposed model may be a promising tool for assessing cerebral hemodynamics in patients with cerebrovascular disease.

Sediment resuspension may play a major role in sediment-water exchange of nutrients, matter and energy in coastal areas where waves and currents dominate sediment transport. Biogeochemical sediment properties regulate sediment erodibility, but there is only limited knowledge of how temporal variability in environmental variables is reflected in the resuspension potential, especially for subtidal habitats. Further, the significance of resuspension on nutrient fluxes in coastal environments has remained unclear as contradicting results have been reported. Here we quantified the temporal variation in resuspension potential metrics (erosion threshold (τc; N m−2) and erosion constant (me; g N−1 s−1)) and associated nutrient fluxes from three sites in the Hanko archipelago (Finland) using a core-based erosion device (EROMES). The sites were sampled bi-monthly from April to December. We also quantified the temporal variation in biogeochemical sediment properties at each site. The τc exhibited the clearest temporal pattern in muddy sediment, where the coefficient of variation (= 67) was two to three times higher than the mixed (= 29) and sandy (= 16) sediments. Dry bulk density was the best predictor for sediment erodibility at all sites explaining 26–46% of the temporal variation in τc despite its limited variability at sandier sites. In addition, temporal variations in the macrofaunal community were important predictors of muddy sediment erodibility and therefore community dynamics need to be considered in sediment transport studies. All sites were potential nutrient sources, yet the overall role of sediment resuspension on nutrient release from the sediments was small.

Objective: Revision using distal inflow (RUDI) is currently proposed in patients on hemodialysis having a high flow access (HFA; >2 L/min) or hemodialysis access-induced distal ischemia (HAIDI). However, a recurrence of high flow or hand ischemia is not unusual in the years after RUDI. The aim of the present study was to describe changes in flow characteristics and arterial diameters in the dialysis arm after RUDI for HFA.Methods: Volume flow, diameter, peak systolic velocity and end diastolic velocity of the brachial artery (BA) were studied 2 and 12 months after RUDI using duplex imaging. In a portion of patients, these characteristics were also assessed at proximal and distal portions of radial and ulnar arteries (proximal forearm radial artery, distal radial artery, ulnar artery, and distal ulnar artery), and in the greater saphenous venous interponate. HFA patients were grouped according to presence of concomitant hand ischemia (HFA-HAIDI) or absence (HFA).Results: Fifteen patients (54 +/- 16 year old; 10 males; HFA-HAIDI, n = 6; HFA, n = 9) with a BA HFA (flow volume, 2740 6 322 mL/min) undergoing RUDI were studied between March 2011 and October 2016 in two Dutch hospitals. After 2 months, flow volume had decreased (1180 +/- 189 mL/min), but again increased at 12 months (1520 +/- 217 mL/min; P Conclusions: RUDI for HFA reduction does not reverse BA dilatation, suggesting irreversible structural arterial wall damage possibly contributing to recurrent high flow. Radial artery remodeling is attenuated in HFA patients previously reporting concurrent hand ischemia diminishing the likelihood of high flow recurrence in this subgroup.

Objective: The aim of the study was to observe the natural haemodynamic changes after arteriovenous fistula (AVF) creation in haemodialysis patients with and without a previous ipsilateral vascular access.Methods: This was a retrospective, single centre cohort study. Patient demographics were registered and preand post-operative vessel ultrasound examinations were performed at regular follow up intervals. Arteriovenous fistula outcomes in terms of vessel diameter and access flow enhancement were determined for radiocephalic, brachiocephalic, and brachiobasilic AVFs.Results: In total, 331 patients (median age 66 years, 60% male) with 366 new autologous AVFs were studied, of whom 112 patients had a previous ipsilateral vascular access (VA). Patients with a previous ipsilateral VA had a statistically significantly greater pre-operative brachial artery diameter (4.4 mm) and flow (106 mL/min), and basilic vein diameter (4.9 mm), compared with patients without a previous ipsilateral VA (4.0 mm, 54 mL/min, and 4.3 mm, respectively). For all AVF configurations these differences gradually disappeared over three months after AVF creation. The haemodynamic changes reached a plateau at three months, and were statistically significantly accelerated in patients with a previous ipsilateral VA. There were no differences in primary failure or high flow complications between both groups.Conclusion: Arteriovenous fistulae show haemodynamic and remodelling changes up to three months post-operatively. Previous ipsilateral VAs may initiate vessel preconditioning, and accelerate the observed haemodynamic changes after AVF creation. However, this preconditioning does not result in a beneficial or detrimental effect on VA function.

Summary Human induced pluripotent stem cells (hiPSCs) are used to study organogenesis and model disease as well as being developed for regenerative medicine. Endothelial cells are among the many cell types differentiated from hiPSCs, but their maturation and stabilization fall short of that in adult endothelium. We examined whether shear stress alone or in combination with pericyte co-culture would induce flow alignment and maturation of hiPSC-derived endothelial cells (hiPSC-ECs) but found no effects comparable with those in primary microvascular ECs. In addition, hiPSC-ECs lacked a luminal glycocalyx, critical for vasculature homeostasis, shear stress sensing, and signaling. We noted, however, that hiPSC-ECs have dysfunctional mitochondrial permeability transition pores, resulting in reduced mitochondrial function and increased reactive oxygen species. Closure of these pores by cyclosporine A improved EC mitochondrial function but also restored the glycocalyx such that alignment to flow took place. These results indicated that mitochondrial maturation is required for proper hiPSC-EC functionality., Graphical Abstract, Highlights • hiPSC-ECs lack a functional glycocalyx and fail to align to flow • hiPSC-ECs have reduced mitochondrial function and increased leakage of ROS • Closing the mPTP with cyclosporine A induces mitochondrial maturation • Improved mitochondrial function restores the glycocalyx and alignment to flow, Closure of the mitochondrial permeability transition pore by cyclosporine A improved mitochondrial function and maturation of hiPSC-ECs but also restored the glycocalyx such that alignment to flow took place. This functional glycocalyx, necessary for growth factor signaling and anticoagulation, is a prerequisite for future hiPSC-EC applications in tissue engineering, organoid vascularization and therapeutic use of hiPSC-ECs.

Objective: The purpose of this study is to assess the value of an automated model-based plaque characterization tool for the prediction of major adverse cardiac events (MACE).Methods: We retrospectively included 45 patients with suspected coronary artery disease of which 16 (33%) experienced MACE within 12 months. Commercially available plaque quantification software was used to automatically extract quantitative plaque morphology: lumen area, wall area, stenosis percentage, wall thickness, plaque burden, remodeling ratio, calcified area, lipid rich necrotic core (LRNC) area and matrix area. The measurements were performed at all cross sections, spaced at 0.5 mm, based on fully 3D segmentations of lumen, wall, and each tissue type. Discriminatory power of these markers and traditional risk factors for predicting MACE were assessed.Results: Regression analysis using clinical risk factors only resulted in a prognostic accuracy of 63% with a corresponding area under the curve (AUC) of 0.587. Based on our plaque morphology analysis, minimal cap thickness, lesion length, LRNC volume, maximal wall area/thickness, the remodeling ratio, and the calcium volume were included into our prognostic model as parameters. The use of morphologic features alone resulted in an increased accuracy of 77% with an AUC of 0.94. Combining both clinical risk factors and morphological features in a multivariate logistic regression analysis increased the accuracy to 87% with a similar AUC of 0.924.Conclusion: An automated model based algorithm to evaluate CCTA-derived plaque features and quantify morphological features of atherosclerotic plaque increases the ability for MACE prognostication significantly compared to the use of clinical risk factors alone.

Gravity-sensitive cellular responses are regularly observed in both specialized and nonspecialized cells. One potential mechanism for this sensitivity is a changing viscosity of the intracellular organelles. Here, we report a novel, to our knowledge, viscosity-sensitive molecular rotor based on mesosubstituted boron-dipyrrin used to investigate the response of viscosity of cellular membranes to hypergravity conditions created at the large diameter centrifuge at the European Space Agency Technology Centre. Mouse osteoblastic (MC3T3-E1) and endothelial (human umbilical vein endothelial cell) cell lines were tested, and an increase in viscosity was found with increasing hypergravity loading. This response is thought to be primarily biologically driven, with the potential for a small, instantaneous physical mechanism also contributing to the observed effect. This work provides the first, to our knowledge, quantitative data for cellular viscosity changes under hypergravity, up to 15 × g.

Objective: Customised aortic repair (CAR) is a new and minimally invasive technique for the endovascular treatment of abdominal aortic aneurysms (AAAs). The aneurysm is completely sealed with a non-contained, non-cross linked polymer, while a new flow lumen is created with balloons. For CAR, the haemodynamically most favourable balloon and flow lumen configuration has not been established before; therefore, four flow parameters were assessed in an in vitro model.Methods: Three in vitro balloon configurations were implanted in an in vitro AAA model; a configuration with crossing balloons (CC) and two parallel configurations (PC1 and PC2). These three models were consecutively placed in a flow system that mimics physiological flow conditions. Laser particle imaging velocimetry (PIV) was used to resolve spatial and temporal flow patterns during the cardiac cycle. In house built algorithms were used to analyse the PIV data for the computing of (i) flow velocity; (ii) vorticity; (iii) wall shear stress (WSS); and (iv) time averaged wall shear stress (TAWSS).Results: Suprarenal flow patterns were similar in all models. The CC showed a higher infrarenal velocity than PC1 and PC2 (38 cm/s vs. 23 cm/s vs. 23 cm/s), and a higher vorticity at the crossing of the lumens (CC: 337/s; PC1 127/s; PC2: 112/s). The lowest vorticity was observed in PC2, especially in the infrarenal neck (CC: 200/s; PC1 164/s; PC2: 98/s). Although WSS and TAWSS varied between configurations, values were in the within non-pathological range.Conclusion: The flow lumens created by three balloon configurations used in an in vitro model of CAR have been studied, and resulted in different haemodynamics. The differences in velocity and lower vorticity, especially at the crossing section of the two balloons, showed that PC2 has favourable haemodynamics compared with the CC and PC1. Future research will be focused on the clinical applicability of CAR based on the PC2 design.

Endothelial–mesenchymal transition (EndMT) is a form of endothelial dysfunction wherein endothelial cells acquire a mesenchymal phenotype and lose endothelial functions, which contributes to the pathogenesis of intimal hyperplasia and atherosclerosis. The mitogen activated protein kinase 7 (MAPK7) inhibits EndMT and decreases the expression of the histone methyltransferase Enhancer-of-Zeste homologue 2 (EZH2), thereby maintaining endothelial quiescence. EZH2 is the catalytic subunit of the Polycomb Repressive Complex 2 that methylates lysine 27 on histone 3 (H3K27me3). It is elusive how the crosstalk between MAPK7 and EZH2 is regulated in the endothelium and if the balance between MAPK7 and EZH2 is disturbed in vascular disease. In human coronary artery disease, we assessed the expression levels of MAPK7 and EZH2 and found that with increasing intima/media thickness ratio, MAPK7 expression decreased, whereas EZH2 expression increased. In vitro, MAPK7 activation decreased EZH2 expression, whereas endothelial cells deficient of EZH2 had increased MAPK7 activity. MAPK7 activation results in increased expression of microRNA (miR)-101, a repressor of EZH2. This loss of EZH2 in turn results in the increased expression of the miR-200 family, culminating in decreased expression of the dual-specificity phosphatases 1 and 6 who may repress MAPK7 activity. Transfection of endothelial cells with miR-200 family members decreased the endothelial sensitivity to TGFβ1-induced EndMT. In endothelial cells there is reciprocity between MAPK7 signaling and EZH2 expression and disturbances in this reciprocal signaling associate with the induction of EndMT and severity of human coronary artery disease.

BACKGROUND AND AIM: Portal vein thrombosis (PVT) is a common complication of cirrhosis. The exact pathophysiology remains largely unknown, and treatment with anticoagulants does not lead to recanalization of the portal vein in all patients. A better insight into the structure and composition of portal vein thrombi may assist in developing strategies for the prevention and treatment of PVT.APPROACH AND RESULTS: Sixteen prospectively and 63 retrospectively collected nonmalignant portal vein thrombi from patients with cirrhosis who underwent liver transplantation were included. Histology, immunohistochemistry, and scanning electron microscopy were used to assess structure and composition of the thrombi. Most recent CT scans were reanalyzed for thrombus characteristics. Clinical characteristics were related to histological and radiological findings. All samples showed a thickened, fibrotic tunica intima. Fibrin-rich thrombi were present on top of the fibrotic intima in 9/16 prospective cases and in 21/63 retrospective cases. A minority of the fibrotic areas stained focally positive for fibrin/fibrinogen (16% of cases), von Willebrand factor (VWF; 10%), and CD61 (platelets, 21%), while most of the fibrin-rich areas stained positive for those markers (fibrin/fibrinogen, 100%; VWF, 77%; CD61, 100%). No associations were found between clinical characteristics including estimated thrombus age and use of anticoagulants and presence of fibrin-rich thrombi.CONCLUSION: We demonstrate that PVT in patients with cirrhosis consists of intimal fibrosis with an additional fibrin-rich thrombus in only one-third of cases. We hypothesize that our observations may explain why not all portal vein thrombi in patients with cirrhosis recanalize by anticoagulant therapy.

Scientific reports 11(1), 16800 (2021). doi:10.1038/s41598-021-96397-2, Published by Macmillan Publishers Limited, part of Springer Nature, [London]

Objective: platelets possess not only haemostatic but also inflammatory properties, which combined are thought to play a detrimental role in thromboinflammatory diseases such as acute coronary syndromes and stroke. Phosphodiesterase (PDE) 3 and -5 inhibitors have demonstrated efficacy in secondary prevention of arterial thrombosis, partially mediated by their antiplatelet action. Yet it is unclear whether such inhibitors also affect platelets’ inflammatory functions. Here, we aimed to examine the effect of the PDE3A inhibitor cilostazol and the PDE5 inhibitor tadalafil on platelet function in various aspects of thromboinflammation. Approach and results: cilostazol, but not tadalafil, delayed ex vivo platelet-dependent fibrin formation under whole blood flow over type I collagen at 1000 s−1. Similar results were obtained with blood from Pde3a deficient mice, indicating that cilostazol effects are mediated via PDE3A. Interestingly, cilostazol specifically reduced the release of phosphatidylserine-positive extracellular vesicles (EVs) from human platelets while not affecting total EV release. Both cilostazol and tadalafil reduced the interaction of human platelets with inflamed endothelium under arterial flow and the release of the chemokines CCL5 and CXCL4 from platelets. Moreover, cilostazol, but not tadalafil, reduced monocyte recruitment and platelet-monocyte interaction in vitro. Conclusions: this study demonstrated yet unrecognised roles for platelet PDE3A and platelet PDE5 in platelet procoagulant and proinflammatory responses.

Shear stress imposed on the arterial wall by the flow of blood affects endothelial cell morphology and function. Low magnitude, oscillatory and multidirectional shear stresses have all been postulated to stimulate a pro-atherosclerotic phenotype in endothelial cells, whereas high magnitude and unidirectional or uniaxial shear are thought to promote endothelial homeostasis. These hypotheses require further investigation, but traditional in vitro techniques have limitations, and are particularly poor at imposing multidirectional shear stresses on cells. One method that is gaining increasing use is to culture endothelial cells in standard multi-well plates on the platform of an orbital shaker; in this simple, low-cost, high-throughput and chronic method, the swirling medium produces different patterns and magnitudes of shear, including multidirectional shear, in different parts of the well. However, it has a significant limitation: cells in one region, exposed to one type of flow, may release mediators into the medium that affect cells in other parts of the well, exposed to different flows, hence distorting the apparent relation between flow and phenotype. Here we present an easy and affordable modification of the method that allows cells to be exposed only to specific shear stress characteristics. Cell seeding is restricted to a defined region of the well by coating the region of interest with fibronectin, followed by passivation using passivating solution. Subsequently, the plates can be swirled on the shaker, resulting in exposure of cells to well-defined shear profiles such as low magnitude multidirectional shear or high magnitude uniaxial shear, depending on their location. As before, the use of standard cell-culture plasticware allows straightforward further analysis of the cells. The modification has already allowed the demonstration of soluble mediators, released from endothelium under defined shear stress characteristics, that affect cells located elsewhere in the well. National Medical Research Council (NMRC) Published version The authors gratefully acknowledge a British Heart Foundation project grant (to PDW), a National Medical Research Council Singapore TAAP and DYNAMO Grant (to XW, NMRC/OFLCG/004/2018, NMRC/OFLCG/001/2017), an A*STAR Graduate Scholarship (to KTP), and a British Heart Foundation Center of Research Excellence studentship (to MA).

Coastal wetlands fulfil important functions for biodiversity conservation and coastal protection, which are inextricably linked to typical morphological features like tidal channels. Channel network configurations in turn are shaped by bio-geomorphological feedbacks between vegetation, hydrodynamics and sediment transport. This study investigates the impact of two starkly different recruitment strategies between mangroves (fast/homogenous) and salt marshes (slow/patchy) on channel network properties. We first compare channel networks found in salt marshes and mangroves around the world and then demonstrate how observed channel patterns can be explained by vegetation establishment strategies using controlled experimental conditions. We find that salt marshes are dissected by more extensive channel networks and have shorter over-marsh flow paths than mangrove systems, while their branching patterns remain similar. This finding is supported by our laboratory experiments, which reveal that different recruitment strategies of mangroves and salt marshes hamper or facilitate channel development, respectively. Insights of our study are crucial to understand wetland resilience with rising sea-levels especially under climate-driven ecotone shifts. ispartof: NATURE COMMUNICATIONS vol:13 issue:1 ispartof: location:England status: published

From developmental stages until adulthood, the circulatory system remodels in response to changes in blood flow in order to maintain vascular homeostasis. Remodeling processes can be driven by de novo formation of vessels or angiogenesis, and by the restructuration of already existing vessels, such as vessel enlargement and regression. Notably, vessel enlargement can occur as fast as in few hours in response to changes in flow and pressure. The high plasticity and responsiveness of blood vessels rely on endothelial cells. Changes within the bloodstream, such as increasing shear stress in a narrowing vessel or lowering blood flow in redundant vessels, are sensed by endothelial cells and activate downstream signaling cascades, promoting behavioral changes in the involved cells. This way, endothelial cells can reorganize themselves to restore normal circulation levels within the vessel. However, the dysregulation of such processes can entail severe pathological circumstances with disturbances affecting diverse organs, such as human hereditary telangiectasias. There are different pathways through which endothelial cells react to promote vessel enlargement and mechanisms may differ depending on whether remodeling occurs in the adult or in developmental models. Understanding the molecular mechanisms involved in the fast-adapting processes governing vessel enlargement can open the door to a new set of therapeutical approaches to be applied in occlusive vascular diseases. Therefore, we have outlined here the latest advances in the study of vessel enlargement in physiology and pathology, with a special insight in the pathways involved in its regulation.

The blood-brain barrier (BBB) is a highly selective cellular monolayer unique to the microvasculature of the central nervous system (CNS), and it mediates the communication of the CNS with the rest of the body by regulating the passage of molecules into the CNS microenvironment. Limitation of passage of substances through the BBB is mainly due to tight junctions (TJ) and adherens junctions (AJ) between brain microvascular endothelial cells. The importance of actin filaments and microtubules in establishing and maintaining TJs and AJs has been indicated; however, recent studies have shown that intermediate filaments are also important in the formation and function of cell-cell junctions. The most common intermediate filament protein in endothelial cells is vimentin. Vimentin plays a role in blood-brain barrier permeability in both cell-cell and cell-matrix interactions by affecting the actin and microtubule reorganization and by binding directly to VE-cadherin or integrin proteins. The BBB permeability increases due to the formation of stress fibers and the disruption of VE-cadherin interactions between two neighboring cells in various diseases, disrupting the fiber network of intermediate filament vimentin in different ways. Intermediate filaments may be long ignored key targets in regulation of BBB permeability in health and disease., EuroCellNet COST Action [CA15214]; Scientific and Technological Research Council of Turkey (TUBITAK) [216M542], The supports of EuroCellNet COST Action CA15214 and The Scientific and Technological Research Council of Turkey (TUBITAK) [grant number 216M542] are acknowledged.

Atherosclerotic plaques prone to rupture may cause acute myocardial infarction (MI) but can also heal without causing an event. Certain common histopathological features, including inflammation, a thin fibrous cap, positive remodelling, a large necrotic core, microcalcification, and plaque haemorrhage are commonly found in plaques causing an acute event. Recent advances in imaging techniques have made it possible to detect not only luminal stenosis and overall coronary atherosclerosis burden but also to identify such adverse plaque characteristics. However, the predictive value of identifying individual adverse atherosclerotic plaques for future events has remained poor. In this Position Paper, the relationship between vulnerable plaque imaging and MI is addressed, mainly for non-invasive assessments but also for invasive imaging of adverse plaques in patients undergoing invasive coronary angiography. Dynamic changes in atherosclerotic plaque development and composition may indicate that an adverse plaque phenotype should be considered at the patient level rather than for individual plaques. Imaging of adverse plaque burden throughout the coronary vascular tree, in combination with biomarkers and biomechanical parameters, therefore holds promise for identifying subjects at increased risk of MI and for guiding medical and invasive treatment.

Fluid flow is a powerful morphogenic force during embryonic development. The physical forces created by flowing fluids can either create morphogen gradients or be translated by mechanosensitive cells into biological changes in gene expression. In this Primer, we describe how fluid flow is created in different systems and highlight the important mechanosensitive signalling pathways involved for sensing and transducing flow during embryogenesis. Specifically, we describe how fluid flow helps establish left-right asymmetry in the early embryo and discuss the role of flow of blood, lymph and cerebrospinal fluid in sculpting the embryonic cardiovascular and nervous system. ispartof: DEVELOPMENT vol:147 issue:15 ispartof: location:England status: published

Shear stress induces directed endothelial cell (EC) migration in blood vessels leading to vessel diameter increase and induction of vascular maturation. Other factors, such as EC elongation and interaction between ECs and non-vascular areas are also important. Computational models have previously been used to study collective cell migration. These models can be used to predict EC migration and its effect on vascular remodelling during embryogenesis. We combined live time-lapse imaging of the remodelling vasculature of the quail embryo yolk sac with flow quantification using a combination of micro-Particle Image Velocimetry and computational fluid dynamics. We then used the flow and remodelling data to inform a model of EC migration during remodelling. To obtain the relation between shear stress and velocity in vitro for EC cells, we developed a flow chamber to assess how confluent sheets of ECs migrate in response to shear stress. Using these data as an input, we developed a multiphase, self-propelled particles (SPP) model where individual agents are driven to migrate based on the level of shear stress while maintaining appropriate spatial relationship to nearby agents. These agents elongate, interact with each other, and with avascular agents at each time-step of the model. We compared predicted vascular shape to real vascular shape after 4 hours from our time-lapse movies and performed sensitivity analysis on the various model parameters. Our model shows that shear stress has the largest effect on the remodelling process. Importantly, however, elongation played an especially important part in remodelling. This model provides a powerful tool to study the input of different biological processes on remodelling., Author summary Shear stress is known to play a leading role in endothelial cell (EC) migration and hence, vascular remodelling. Vascular remodelling is, however, more complicated than only EC migration. To achieve a better understanding of this process, we developed a computational model in which, shear stress mediated EC migration has the leading role and other factors, such as avascular regions and EC elongation, are also accounted for. We have tested this model for different vessel shapes during remodelling and could study the role that each of these factors play in remodelling. This model gives us the possibility of addition of other factors such as biochemical signals and angiogenesis which will help us in the study of vascular remodelling in both development and disease.

Bone is one of the most highly adaptive tissues in the body, possessing the capability to alter its morphology and function in response to stimuli in its surrounding environment. The ability of bone to sense and convert external mechanical stimuli into a biochemical response, which ultimately alters the phenotype and function of the cell, is described as mechanotransduction. This review aims to describe the fundamental physiology and biomechanisms that occur to induce osteogenic adaptation of a cell following application of a physical stimulus. Considerable developments have been made in recent years in our understanding of how cells orchestrate this complex interplay of processes, and have become the focus of research in osteogenesis. We will discuss current areas of preclinical and clinical research exploring the harnessing of mechanotransductive properties of cells and applying them therapeutically, both in the context of fracture healing and de novo bone formation in situations such as nonunion.Cite this article: Bone Joint Res 2019;9(1):1–14.

Stem cell expansion on 3D porous scaffolds cultured in bioreactor systems has been shown to be beneficial for maintenance of the original cell functionality in tissue engineering strategies (TE). However, the production of extracellular matrix (ECM) makes harvesting the progenitor cell population from 3D scaffolds a challenge. Medium composition plays a role in stimulating cell proliferation over extracellular matrix (ECM) production. In this regard, a computational model describing tissue growth inside 3D scaffolds can be a great tool in designing optimal experimental conditions. In this study, a computational model describing cell and ECM growth in a perfusion bioreactor is developed, including a description of the effect of a (generic) growth factor on the biological processes taking place inside the 3D scaffold. In the model, the speed of cell and ECM growth depends on the flow-induced shear stress, curvature and the concentrations of oxygen, glucose, lactate, and growth factor. The effect of the simulated growth factor is to differentially enhance cell proliferation over ECM production. After model calibration with historic in-house data, a multi-objective optimization procedure is executed aiming to minimize the total experimental cost whilst maximizing cell growth during culture. The obtained results indicate there are multiple optimum points for the medium refreshment regime and the initial growth factor concentration where a trade-off is made between the final amount of cells and the culture cost. Finally, the model is applied to experiments reported in the literature studying the effects of perfusion-based cell culture and/or growth factor supplementation on cell expansion. The qualitative similarities between the simulation and experimental results, even in the absence of proper model calibration, reinforces the generic character of the proposed modeling framework. The model proposed in this study can contribute to the cost efficient production of cell-based TE products, ultimately contributing to their affordability and accessibility. ispartof: FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY vol:8 ispartof: location:Switzerland status: published

Background: The incidence of atherosclerotic cardiovascular diseases, which constitute an important disease group, is expected to rise with increasing life expectancy. The aim of this study was to investigate the predictive values of triglyceride/HDL cholesterol (TG/HDL-C) ratio and whole blood viscosity on possible severe carotid or peripheral arterial disease in patients who were admitted to our clinic and scheduled for coronary artery bypass graft (CABG) surgery. Methods: This study retrospectively examined 299 patients who were admitted to our clinic and scheduled for CABG between August 1, 2015, and August 1, 2019. Patients with severe carotid or peripheral arterial disease were included in group 2; those in whom peripheral arterial disease and stenosis were not detected were included in group 1. Results: Group 1 consisted of 255 patients, mean (+/- standard deviation) age 58.1 +/- 8.9 years; the 44 patients in group 2 had a mean age of 64.1 +/- 10.6 years (P = .018). The incidence of diabetes mellitus and tobacco use were significantly higher in group 2 (P < .001 and P = .034, respectively). Triglyceride values, TG/HDL-C, and whole blood viscosity at high and low shear rates were significantly higher in group 2 (P = .017, P < .001, P < .001, and P < .001, respectively). Receiver operator characteristic analysis revealed that the cutoff value of TG/HDL-C was 3.2 with 77% sensitivity and 72% specificity; that of high-shear-rate viscosity was 16.1 with 73% sensitivity and 69% specificity; and that of low-shear-rate viscosity was 59.2 with 70% sensitivity and 63% specificity. Conclusion: It is possible to predict severe additional arterial diseases in patients scheduled for CABG with whole blood viscosity and TG/HDL-C ratios, both of which can be obtained with simple blood tests.

Recent studies suggest that substituting sitting with light physical activity has beneficial metabolic effects, but it is unclear if this is associated with parallel changes in endothelial function. Data from three randomized cross-over studies were analyzed, in which 61 subjects (with normal weight, overweight and type 2 diabetes) followed different activity regimens (Sit, SitLess and/or Exercise) of four days each. Subjects were instructed to sit 14 h/day ('Sit'), to substitute 1 h/day of sitting with moderate-to-vigorous cycling ('Exercise') or to substitute 5-6 h/day sitting with light-intensity walking and standing ('SitLess'). Physical activity was assessed 24 h/day by accelerometry (ActivPAL) and diet was standardized. Fasted circulating biomarkers of endothelial dysfunction, lipids and insulin sensitivity were assessed the morning after each activity regimen. The endothelial dysfunction score (ED-score) was computed by averaging the Z-scores of the circulating biomarkers of endothelial dysfunction. Compared to Sit, Exercise resulted in lower ED-score, sICAM1 and sE-selectin (p