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Midlife hypertension is an important risk factor for cognitive impairment and dementia, including Alzheimer's disease. We investigated the effects of long-term treatment with two classes of antihypertensive drugs to determine whether diverging mechanisms of blood pressure lowering impact the brain differently. Spontaneously hypertensive rats (SHR) were either left untreated or treated with a calcium channel blocker (amlodipine) or beta blocker (atenolol) until one year of age. The normotensive Wistar Kyoto rat (WKY) was used as a reference group. Both drugs lowered blood pressure equally, while only atenolol decreased heart rate. Cerebrovascular resistance was increased in SHR, which was prevented by amlodipine but not atenolol. SHR showed a larger carotid artery diameter with impaired pulsatility, which was prevented by atenolol. Cerebral arteries demonstrated inward remodelling, stiffening and endothelial dysfunction in SHR. Both treatments similarly improved these parameters. MRI revealed that SHR have smaller brains with enlarged ventricles. In addition, neurofilament light levels were increased in cerebrospinal fluid of SHR. However, neither treatment affected these parameters. In conclusion, amlodipine and atenolol both lower blood pressure, but elicit a different hemodynamic profile. Both medications improve cerebral artery structure and function, but neither drug prevented indices of brain damage in this model of hypertension., (© 2023. The Author(s).)

There is a discrepancy between successful recanalization and good clinical outcome after endovascular treatment (EVT) in acute ischemic stroke patients. During removal of a thrombus, a shower of microemboli may release and lodge to the distal circulation. The objective of this study was to determine the extent of damage on brain tissue caused by microemboli. In a rat model of microembolization, a mixture of microsphere (MS) sizes (15, 25 and 50 µm diameter) was injected via the left internal carotid artery. A 3D image of the left hemisphere was reconstructed and a point-pattern spatial analysis was applied based on G- and K-functions to unravel the spatial correlation between MS and the induced hypoxia or infarction. We show a spatial correlation between MS and hypoxia or infarction spreading up to a distance of 1000-1500 µm. These results imply that microemboli, which individually may not always be harmful, can interact and result in local areas of hypoxia or even infarction when lodged in large numbers., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: Endovascular treatment (EVT) has become the standard of care for acute ischemic stroke. Despite successful recanalization, a limited subset of patients benefits from the new treatment. Human MRI studies have shown that during removal of the thrombus, a shower of microclots is released from the initial thrombus, possibly causing new ischemic lesions. The aim of the current study is to quantify tissue damage following microembolism., Materials and Methods: In a rat model, microembolism was generated by injection of a mixture of polystyrene fluorescent microspheres (15, 25 and 50 µm in diameter). The animals were killed at three time-points: day 1, 3 or 7. AMIRA and IMARIS software was used for 3D reconstruction of brain structure and damage, respectively., Conclusions: Microembolism induces ischemia, hypoxia and infarction. Infarcted areas persist, but hypoxic regions recover over time suggesting that repair processes in the brain rescue the regions at risk., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The hippocampus is susceptible to protein aggregation in neurodegenerative diseases such as Alzheimer's disease. This protein accumulation is partially attributed to an impaired clearance; however, the removal pathways for fluids and waste products are not fully understood. The aim of this study was therefore to map the clearance pathways from the mouse brain. A mixture of two fluorescently labeled tracers with different molecular weights was infused into the hippocampus. A small subset of mice ( n = 3) was sacrificed directly after an infusion period of 10 min to determine dispersion of the tracer due to the infusion, while another group was sacrificed after spreading of the tracers for an additional 80 min ( n = 7). Upon sacrifice, mice were frozen and sectioned as a whole by the use of a custom-built automated imaging cryomicrotome. Detailed 3D reconstructions were created to map the tracer spreading. We observed that tracers distributed over the hippocampus and entered adjacent brain structures, such as the cortex and cerebroventricular system. An important clearance pathway was found along the ventral part of the hippocampus and its bordering interpeduncular cistern. From there, tracers left the brain via the subarachnoid spaces in the directions of both the nose and the spinal cord. Although both tracers followed the same route, the small tracer distributed further, implying a major role for diffusion in addition to convection. Taken together, these results reveal an important clearance pathway of solutes from the hippocampus., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Naessens, Dobbe, de Vos, VanBavel and Bakker.)

Background: Proper neuronal function is directly dependent on the composition, turnover, and amount of interstitial fluid that bathes the cells. Most of the interstitial fluid is likely to be derived from ion and water transport across the brain capillary endothelium, a process that may be altered in hypertension due to vascular pathologies as endothelial dysfunction and arterial remodelling. In the current study, we investigated the effects of hypertension on the brain for differences in the water homeostasis., Methods: Magnetic resonance imaging (MRI) was performed on a 7T small animal MRI system on male spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) of 10 months of age. The MRI protocol consisted of T2-weighted scans followed by quantitative apparent diffusion coefficient (ADC) mapping to measure volumes of different anatomical structures and water diffusion respectively. After MRI, we assessed the spatial distribution of aquaporin 4 expression around blood vessels., Results: MRI analysis revealed a significant reduction in overall brain volume and remarkably higher cerebroventricular volume in SHR compared to WKY. Whole brain ADC, as well as ADC values of a number of specific anatomical structures, were significantly lower in hypertensive animals. Additionally, SHR exhibited higher brain parenchymal water content. Immunohistochemical analysis showed a profound expression of aquaporin 4 around blood vessels in both groups, with a significantly larger area of influence around arterioles. Evaluation of specific brain regions revealed a decrease in aquaporin 4 expression around capillaries in the corpus callosum of SHR., Conclusion: These results indicate a shift in the brain water homeostasis of adult hypertensive rats.

In sickle cell disease (SCD), oxygen delivery is impaired due to anemia, especially during times of increased metabolic demand, and cerebral blood flow (CBF) must increase to meet changing physiologic needs. But hyperemia limits cerebrovascular reserve (CVR) and ischemic risk prevails despite elevated CBF. The cerebral metabolic rate of oxygen (CMRO 2 ) directly reflects oxygen supply and consumption and may therefore be more insightful than flow-based CVR measures for ischemic risk in SCD. We hypothesized that adults with SCD have impaired CMRO 2 at rest and that a vasodilatory challenge with acetazolamide would improve CMRO 2 . CMRO 2 was calculated from CBF and oxygen extraction fraction (OEF), measured with arterial spin labeling and T 2 -prepared tissue relaxation with inversion recovery (T 2 -TRIR) MRI. We studied 36 adults with SCD without a clinical history of overt stroke, and nine healthy controls. As expected, CBF was higher in patients with SCD versus controls (mean ± SD: 74 ± 16 versus 46 ± 5 mL/100 g/min, P < .001), resulting in similar oxygen delivery (SCD: 377 ± 67 versus controls: 368 ± 42 μmol O 2 /100g/min, P = .69). OEF was lower in patients versus controls (27 ± 4 versus 35 ± 4%, P < .001), resulting in lower CMRO 2 in patients versus controls (102 ± 24 versus 127 ± 20 μmol O 2 /100g/min, P = .002). After acetazolamide, CMRO 2 declined further in patients (P < .01) and did not decline significantly in controls (P = .78), indicating that forcing higher CBF worsened oxygen utilization in SCD patients. This lower CMRO 2 could reflect variation between healthy and unhealthy vascular beds in terms of dilatory capacity and resistance whereby dysfunctional vessels become more oxygen-deprived, hence increasing the risk of localized ischemia., (© 2020 The Authors. American Journal of Hematology published by Wiley Periodicals, Inc.)

Background: Many processes contributing to the functional and structural regulation of the coronary circulation have been identified. A proper understanding of the complex interplay of these processes requires a quantitative systems approach that includes the complexity of the coronary network. The purpose of this study was to provide a detailed quantification of the branching characteristics and local hemodynamics of the human coronary circulation., Methods: The coronary arteries of a human heart were filled post-mortem with fluorescent replica material. The frozen heart was alternately cut and block-face imaged using a high-resolution imaging cryomicrotome. From the resulting 3D reconstruction of the left coronary circulation, topological (node and loop characteristics), topographic (diameters and length of segments), and geometric (position) properties were analyzed, along with predictions of local hemodynamics (pressure and flow)., Results: The reconstructed left coronary tree consisted of 202,184 segments with diameters ranging from 30 μm to 4 mm. Most segments were between 100 μm and 1 mm long. The median segment length was similar for diameters ranging between 75 and 200 μm. 91% of the nodes were bifurcations. These bifurcations were more symmetric and less variable in smaller vessels. Most of the pressure drop occurred in vessels between 200 μm and 1 mm in diameter. Downstream conductance variability affected neither local pressure nor median local flow and added limited extra variation of local flow. The left coronary circulation perfused 358 cm 3 of myocardium. Median perfused volume at a truncation level of 100 to 200 μm was 20 mm 3 with a median perfusion of 5.6 ml/min/g and a high local heterogeneity., Conclusion: This study provides the branching characteristics and hemodynamic analysis of the left coronary arterial circulation of a human heart. The resulting model can be deployed for further hemodynamic studies at the whole organ and local level., (Copyright © 2020 Schwarz, van Lier, van den Wijngaard, Siebes and VanBavel.)

New Findings: What is the topic of this review? In this symposium report, we review the glymphatic clearance from the brain. What advances does it highlight? Evaluation of the evidence indicates that cerebrospinal fluid flows along paravascular spaces at the surface of the brain. However, bulk flow along penetrating arteries into the brain, followed by exit along veins, requires further confirmation. Clearance from the brain, based on mixing, might provide an alternative explanation for experimental findings., Abstract: The interstitial fluid of the brain provides the environment for proper neuronal function. Maintenance of the volume and composition of interstitial fluid requires regulation of the influx and removal of water, ions, nutritive and waste products. The recently described glymphatic pathway might contribute to some of these functions. It is proposed that cerebrospinal fluid enters the brain via paravascular spaces along arteries, mixes with interstitial fluid, and leaves the brain via paravascular spaces along veins. In this symposium report, we review the glymphatic concept, its concerns, and alternative views on interstitial fluid-cerebrospinal fluid exchange., (© 2018 The Authors Experimental Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.)

Background and Purpose- We developed a rat model of silent brain infarcts based on microsphere infusion and investigated their impact on perfusion and tissue damage. Second, we studied the extent and mechanisms of perfusion recovery. Methods- At day 0, 15 µm fluorescent microspheres were injected into the right common carotid artery of F344 rats. At days 1, 7, or 28, the brain was removed, cut in 100-µm cryosections, and processed for immunofluorescent staining and analysis. Results- Injection of microspheres caused mild and transient damage to the treated hemisphere, with a decrease in perfused capillary volume at day 1, as compared with the untreated hemisphere. At day 1 but not at days 7 and 28, we observed IgG staining outside of the vessels, indicating vessel leakage. All microspheres were located inside the lumen of the vessels at day 1, whereas the vast majority (≈80%) of the microspheres were extravascular at day 7, and 100% at day 28. This was accompanied by restoration of perfused capillary volume. Conclusions- Microspheres cause mild and transient damage, and effective extravasation mechanisms exist in the brain to clear microsized emboli from the vessels.

Sickle cell disease is characterized by chronic hemolytic anemia and vascular inflammation, which can diminish the vasodilatory capacity of the small resistance arteries, making them less adept at regulating cerebral blood flow. Autoregulation maintains adequate oxygen delivery, but when vasodilation is maximized, the low arterial oxygen content can lead to ischemia and silent cerebral infarcts. We used magnetic resonance imaging of cerebral blood flow to quantify whole-brain cerebrovascular reserve in 36 adult patients with sickle cell disease (mean age, 31.9±11.3 years) and 11 healthy controls (mean age, 37.4±15.4 years), and we used high-resolution 3D FLAIR magnetic resonance imaging to determine the prevalence of silent cerebral infarcts. Cerebrovascular reserve was calculated as the percentage change in cerebral blood flow after a hemodynamic challenge with acetazolamide. Co-registered lesion maps were used to demonstrate prevalent locations for silent cerebral infarcts. Cerebral blood flow was elevated in patients with sickle cell disease compared to controls (median [interquartile range]: 82.8 [20.1] vs 51.3 [4.8] mL/100g/min, P <0.001). Cerebral blood flow was inversely associated with age, hemoglobin, and fetal hemoglobin, and correlated positively with bilirubin, and LDH, indicating that cerebral blood flow may reflect surrogates of hemolytic rate. Cerebrovascular reserve in sickle cell disease was decreased by half compared to controls (34.1 [33.4] vs 69.5 [32.4] %, P <0.001) and was associated with hemoglobin and erythrocyte count indicating anemia-induced hemodynamic adaptations. In total, 29/36 patients (81%) and 5/11 controls (45%) had silent cerebral infarcts (median volume of 0.34 vs 0.02 mL, P =0.03). Lesions were preferentially located in the borderzone. In conclusion, patients with sickle cell disease have a globally reduced cerebrovascular reserve as determined by arterial spin labeling with acetazolamide and reflects anemia-induced impaired vascular function in sickle cell disease. This study was registered at clinicaltrials.gov identifier 02824406 ., (Copyright© 2019 Ferrata Storti Foundation.)

Background: Drift is a well-known issue affecting intracoronary pressure measurements. A small pressure offset at the end of the procedure is generally considered acceptable, while repeat assessment is advised for drift exceeding ±2 mmHg. This practice implies that drift assessed after wire pullback equals that at the time of stenosis appraisal, but this assumption has not been systematically investigated. Our aim was to compare intra-and post-procedural pressure sensor drift and assess benefits of correction for intra-procedural drift and its effect on diagnostic classification., Methods: In 70 patients we compared intra- and post-procedural pressure drift for 120 hemodynamic tracings obtained at baseline and throughout the hyperemic response to intracoronary adenosine. Intra-procedural drift was derived from the intercept of the stenosis pressure gradient-velocity relationship. Diagnostic reclassification after correction for intra-procedural drift was assessed for the mean distal-to-aortic pressure ratio at baseline (Pd/Pa) and hyperemia (fractional flow reserve, FFR), and corresponding stenosis resistances., Results: Post- and intra-procedural drift exceeding the tolerated threshold was observed in 73% and 64% of the hemodynamic tracings, respectively. Discordance in terms of acceptable drift level was present for 42% of the tracings, with avoidable repeat physiological assessment in 25% and unacceptable intra-procedural drift unrecognized at final drift check in 17% of the tracings. Correction for intra-procedural drift caused higher reclassification rates for baseline than hyperemic functional indices., Conclusions: Post-procedural pressure drift frequently does not match drift during physiological assessment. Tracing-specific correction for intra-procedural drift can potentially lower the risk of inadvertent diagnostic misclassification and prevent unnecessary repeats., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Objectives: Minimally invasive aortic valve replacement has proven its value over the last decade by its significant advancement and reduction in mortality, morbidity and admission time. However, minimally invasive aortic valve replacement is associated with some on-site difficulties such as limited aortic annulus exposure. Currently, computed tomography scans are used to evaluate the anatomical relationship among the intercostal spaces, ascending aorta and aortic valve prior to surgery. We hypothesized that quantitative measurements of access distance and access angle are associated with outcome and access difficulty., Methods: We introduce a novel minimally invasive aortic valve replacement planning prototype that allows automatic measurements of access angle, access distance and aortic annulus dimensions. The prototype visualizes these measurements on the chest cage as ISO contours. The association of these measures with outcome parameters such as extracorporeal circulation time, aortic cross-clamping time and access difficulty score was assessed. We included 14 patients who received a new valve by ministernotomy., Results: The mean access angle was 40.3 ± 5.1°. It was strongly associated with aortic cross-clamping time (Pearson correlation coefficient = 0.60, P = 0.02) and access difficulty score (Spearman's rank correlation coefficient = 0.57, P = 0.03). Access angles were significantly different between easy and difficult access groups (P = 0.03). There was no significant association between access distance and outcome parameters., Conclusions: Access angle is strongly associated with procedure complexity. The automated presentation of this measure suggests added value of the prototype in clinical practice.

Background: Hypertension is an important risk factor for cerebrovascular disease, including stroke and dementia. Both in humans and animal models of hypertension, neuropathological features such as brain atrophy and oedema have been reported. We hypothesised that cerebrovascular damage resulting from chronic hypertension would manifest itself in a more permeable blood-brain barrier and blood-cerebrospinal fluid barrier. In addition, more leaky barriers could potentially contribute to an enhanced interstitial fluid and cerebrospinal fluid formation, which could, in turn, lead to an elevated intracranial pressure., Methods: To study this, we monitored intracranial pressure and estimated the cerebrospinal fluid production rate in spontaneously hypertensive (SHR) and normotensive rats (Wistar Kyoto, WKY) at 10 months of age. Blood-brain barrier and blood-cerebrospinal fluid barrier integrity was determined by measuring the leakage of fluorescein from the circulation into the brain and cerebrospinal fluid compartment. Prior to sacrifice, a fluorescently labelled lectin was injected into the bloodstream to visualise the vasculature and subsequently study a number of specific vascular characteristics in six different brain regions., Results: Blood and brain fluorescein levels were not different between the two strains. However, cerebrospinal fluid fluorescein levels were significantly lower in SHR. This could not be explained by a difference in cerebrospinal fluid turnover, as cerebrospinal fluid production rates were similar in SHR and WKY, but may relate to a larger ventricular volume in the hypertensive strain. Also, intracranial pressure was not different between SHR and WKY. Morphometric analysis of capillary volume fraction, number of branches, capillary diameter, and total length did not reveal differences between SHR and WKY., Conclusion: In conclusion, we found no evidence for blood-brain barrier or blood-cerebrospinal fluid barrier leakage to a small solute, fluorescein, in rats with established hypertension.

Clearance of waste products from the brain is of vital importance. Recent publications suggest a potential clearance mechanism via paravascular channels around blood vessels. Arterial pulsations might provide the driving force for paravascular flow, but its flow pattern remains poorly characterized. In addition, the relationship between paravascular flow around leptomeningeal vessels and penetrating vessels is unclear. In this study, we determined blood flow and diameter pulsations through a thinned-skull cranial window. We observed that microspheres moved preferentially in the paravascular space of arteries rather than in the adjacent subarachnoid space or around veins. Paravascular flow was pulsatile, generated by the cardiac cycle, with net antegrade flow. Confocal imaging showed microspheres distributed along leptomeningeal arteries, while their presence along penetrating arteries was limited to few vessels. These data suggest that paravascular spaces around leptomeningeal arteries form low resistance pathways on the surface of the brain that facilitate cerebrospinal fluid flow.

Stroke risk in children with sickle cell disease (SCD) is currently assessed with routine transcranial Doppler ultrasound (TCD) measurements of blood velocity in the Circle of Willis (CoW). However, there is currently no biomarker with proven prognostic value in adult patients. Four-dimensional (4D) flow magnetic resonance imaging (MRI) may improve risk profiling based on intracranial haemodynamics. We conducted neurovascular 4D flow MRI and blood sampling in 69 SCD patients [median age 15 years (interquartile range, IQR: 12-50)] and 14 healthy controls [median age 21 years (IQR: 18-43)]. We measured velocity, flow, lumen area and endothelial shear stress (ESS) in the CoW. SCD patients had lower haematocrit and viscosity, and higher velocity, flow and lumen area, with lower ESS compared to healthy controls. We observed significant age-related decline in haemodynamic 4D flow parameters; velocity (Spearman's ρ = -0·36 to -0·61), flow (ρ = -0·26 to -0·52) and ESS (ρ = -0·14 to -0·54) in SCD patients. Further analysis in only adults showed that velocity values were similar in SCD patients compared to healthy controls, but that the additional 4D flow parameters, flow and lumen area, were higher, and ESS lower, in the SCD group. Our data suggest that 4D flow MRI may identify adult patients with an increased stroke risk more accurately than current TCD-based velocity., (© 2017 John Wiley & Sons Ltd.)

Baseline assessment of functional stenosis severity has been proposed as a practical alternative to hyperemic indices. However, intact autoregulation mechanisms may affect intracoronary hemodynamics. The aim of this study was to investigate the effect of changes in aortic pressure (Pa) and heart rate (HR) on baseline coronary hemodynamics and functional stenosis assessment. In 15 patients (55 ± 3% diameter stenosis) Pa, intracoronary pressure (Pd) and flow velocity were obtained at control, and during atrial pacing at 120 bpm, increased Pa (+30 mmHg) with intravenous phenylephrine (PE), and elevated Pa while pacing at sinus heart rate (PE + sHR). We derived rate pressure product (RPP = systolic Pa × HR), baseline microvascular resistance (BMR = Pd/velocity), and stenosis resistance [BSR = (Pa - Pd)/velocity] as well as whole-cycle Pd/Pa. Tachycardia (120 ± 1 bpm) raised RPP by 74% vs., Control: Accordingly, BMR decreased by 27% (p < 0.01) and velocity increased by 36% (p < 0.05), while Pd/Pa decreased by 0.05 ± 0.02 (p < 0.05) and BSR remained similar to control. Raising Pa to 121 ± 3 mmHg (PE) with concomitant reflex bradycardia increased BMR by 26% (p < 0.001) at essentially unchanged RPP and velocity. Consequently, BSR and Pd/Pa were only marginally affected. During PE + sHR, velocity increased by 21% (p < 0.01) attributable to a 46% higher RPP (p < 0.001). However, BMR, BSR, and Pd/Pa remained statistically unaffected. Nonetheless, the interventions tended to increase functional stenosis severity, causing Pd/Pa and BSR of borderline lesions to cross the diagnostic threshold. In conclusion, coronary microvascular adaptation to physiological conditions affecting metabolic demand at rest influences intracoronary hemodynamics, which may lead to altered basal stenosis indices used for clinical decision-making.

Background: Transcatheter aortic valve implantation (TAVI) is a well-established treatment for patients with severe aortic valve stenosis. This procedure requires pre-operative planning by assessment of aortic dimensions on CT Angiography (CTA). It is well-known that the aortic root dimensions vary over the heart cycle. However, sizing is commonly performed at either mid-systole or end-diastole only, which has resulted in an inadequate understanding of its full dynamic behavior., Study Goal: We studied the variation in annulus measurements during the cardiac cycle and determined if this variation is dependent on the amount of calcification at the annulus., Methods: We measured and compared aortic root annular dimensions and calcium volume in CTA acquisitions at 10 cardiac cycle phases in 51 aortic stenosis patients. Sub-group analysis was performed based on the volume of calcium by splitting the population into mildly and severely calcified valves subgroups., Results: For most annulus measurements, the largest differences were found between 10% and 70 to 80% cardiac cycle phases. Mean difference (±standard deviation) in annular minimum diameter, maximum diameter, area, and aspect ratio between mid-systole and end-diastole phases were 1.0 ± 0.29 mm (p = 0.065), 0.30 ± 0.24 mm (p = 0.7), 24.1 ± 7.6 mm2 (p < 0.001), and 0.041 ± 0.012 (p = 0.039) respectively. Calcium volume measurements varied strongly during the cardiac cycle. The dynamic annulus area was behaving differently between mildly and severely calcified subgroups (p = 0.02). Furthermore, patients with severe aortic calcification were associated with larger annulus diameters., Conclusion: There is a significant variation of annulus area and calcium volume measurement during the cardiac cycle. In our measurements, only the dynamic variation of the annulus area is dependent on the severity of the aortic calcification. For TAVI candidates, the annulus area is significantly larger in mid-systole compared to end-diastole.

Hypertension is associated with cognitive decline and various forms of dementia, including Alzheimer's disease. In animal models of hypertension, many of Alzheimer's disease characteristics are recapitulated, including brain atrophy, cognitive decline, amyloid β accumulation and blood brain barrier dysfunction. Removal of amyloid β and other waste products depends in part on clearance via the brain interstitial fluid (ISF). Here we studied the impact of hypertension on ISF drainage, using spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). At 8 months, high (500 kD) and low (3 kD) fluorescent molecular weight tracers released passively into the hippocampus showed a drastically enhanced spreading in SHR. Tracer spreading was inhomogeneous, with accumulation at ISF-CSF borders, around arteries, and towards the stratum lacunosum moleculare. These locations stained positively for the astrocyte marker GFAP, and aquaporin 4. Despite enhanced dispersion, clearance of tracers was not affected in SHR. In conclusion, these data indicate enhanced bulk flow of ISF in the hippocampus of hypertensive rats. ISF drains along astrocytes towards the cerebrospinal fluid compartment, which leads to sieving of high molecular weight solutes. Sieving may lead to a local increase in the concentration of waste products and potentially promotes the aggregation of amyloid β.

Recent evidence suggests an extensive exchange of fluid and solutes between the subarachnoid space and the brain interstitium, involving preferential pathways along blood vessels. We studied the anatomical relations between brain vasculature, cerebrospinal fluid compartments, and paravascular spaces in male Wistar rats. A fluorescent tracer was infused into the cisterna magna, without affecting intracranial pressure. Tracer distribution was analyzed using a 3D imaging cryomicrotome, confocal microscopy, and correlative light and electron microscopy. We found a strong 3D colocalization of tracer with major arteries and veins in the subarachnoid space and large cisterns, attributed to relatively large subarachnoid space volumes around the vessels. Confocal imaging confirmed this colocalization and also revealed novel cisternal connections between the subarachnoid space and ventricles. Unlike the vessels in the subarachnoid space, penetrating arteries but not veins were surrounded by tracer. Correlative light and electron microscopy images indicated that this paravascular space was located outside of the endothelial layer in capillaries and just outside of the smooth muscle cells in arteries. In conclusion, the cerebrospinal fluid compartment, consisting of the subarachnoid space, cisterns, ventricles, and para-arteriolar spaces, forms a continuous and extensive network that surrounds and penetrates the rat brain, in which mixing may facilitate exchange between interstitial fluid and cerebrospinal fluid.

Development of collateral vessels, arteriogenesis, may protect against tissue ischemia, however, quantitative data on this process remain scarce. We have developed a technique for replicating the entire arterial network of ischemic rat hindlimbs in three dimensions (3D) based on vascular casting and automated sequential cryo-imaging. Various dilutions of Batson's No. 17 with methyl methacrylate were evaluated in healthy rats, with further protocol optimization in ischemic rats. Penetration of the resin into the vascular network greatly depended on dilution; the total length of casted vessels below 75 µm was 13-fold higher at 50% dilution compared with the 10% dilution. Dilutions of 25-30%, with transient clamping of the healthy iliac artery, were optimal for imaging the arterial network in unilateral ischemia. This protocol completely filled the lumina of small arterioles and collateral vessels. These appeared as thin anastomoses in healthy legs and increasingly larger vessels during ligation (median diameter 1 week: 63 µm, 4 weeks: 127 µm). The presented combination of quality casts with high-resolution cryo-imaging enables automated, detailed 3D analysis of collateral adaptation, which furthermore can be combined with co-registered 3D distributions of fluorescent molecular imaging markers reflecting biological activity or perfusion.

Minimally invasive aortic valve replacement (mini-AVR) procedures are a valuable alternative to conventional open heart surgery. Currently, planning of mini-AVR consists of selection of the intercostal space closest to the sinotubular junction on preoperative computer tomography images. We developed an automated algorithm detecting the sinotubular junction (STJ) and intercostal spaces for finding the optimal incision location. The accuracy of the STJ detection was assessed by comparison with manual delineation by measuring the Euclidean distance between the manually and automatically detected points. In all 20 patients, the intercostal spaces were accurately detected. The median distance between automated and manually detected STJ locations was 1.4 [IQR= 0.91-4.7] mm compared to the interobserver variation of 1.0 [IQR= 0.54-1.3] mm. For 60% of patients, the fourth intercostal space was the closest to the STJ. The proposed algorithm is the first automated approach for detecting optimal incision location and has the potential to be implemented in clinical practice for planning of various mini-AVR procedures., (Copyright © 2016 IPEM. Published by Elsevier Ltd. All rights reserved.)

Introduction: Wall shear stress (WSS) and oscillatory shear index (OSI) are associated with atherosclerotic disease. Both parameters are derived from blood velocities, which can be measured with phase-contrast MRI (PC-MRI). Limitations in spatiotemporal resolution of PC-MRI are known to affect these measurements. Our aim was to investigate the effect of spatiotemporal resolution using a carotid artery phantom., Methods: A carotid artery phantom was connected to a flow set-up supplying pulsatile flow. MRI measurement planes were placed at the common carotid artery (CCA) and internal carotid artery (ICA). Two-dimensional PC-MRI measurements were performed with thirty different spatiotemporal resolution settings. The MRI flow measurement was validated with ultrasound probe measurements. Mean flow, peak flow, flow waveform, WSS and OSI were compared for these spatiotemporal resolutions using regression analysis. The slopes of the regression lines were reported in %/mm and %/100ms. The distribution of low and high WSS and OSI was compared between different spatiotemporal resolutions., Results: The mean PC-MRI CCA flow (2.5±0.2mL/s) agreed with the ultrasound probe measurements (2.7±0.02mL/s). Mean flow (mL/s) depended only on spatial resolution (CCA:-13%/mm, ICA:-49%/mm). Peak flow (mL/s) depended on both spatial (CCA:-13%/mm, ICA:-17%/mm) and temporal resolution (CCA:-19%/100ms, ICA:-24%/100ms). Mean WSS (Pa) was in inverse relationship only with spatial resolution (CCA:-19%/mm, ICA:-33%/mm). OSI was dependent on spatial resolution for CCA (-26%/mm) and temporal resolution for ICA (-16%/100ms). The regions of low and high WSS and OSI matched for most of the spatiotemporal resolutions (CCA:30/30, ICA:28/30 cases for WSS; CCA:23/30, ICA:29/30 cases for OSI)., Conclusion: We show that both mean flow and mean WSS are independent of temporal resolution. Peak flow and OSI are dependent on both spatial and temporal resolution. However, the magnitude of mean and peak flow, WSS and OSI, and the spatial distribution of OSI and WSS did not exhibit a strong dependency on spatiotemporal resolution., Competing Interests: The authors have declared that no competing interests exist.

Despite the importance of collateral vessels in human hearts, a detailed analysis of their distribution within the coronary vasculature based on three-dimensional vascular reconstructions is lacking. This study aimed to classify the transmural distribution and connectivity of coronary collaterals in human hearts. One normotrophic human heart and one hypertrophied human heart with fibrosis in the inferior wall from a previous infarction were obtained. After filling the coronary arteries with fluorescent replica material, hearts were frozen and alternately cut and block-face imaged using an imaging cryomicrotome. Transmural distribution, connectivity, and diameter of collaterals were determined. Numerous collateral vessels were found (normotrophic heart: 12.3 collaterals/cm(3); hypertrophied heart: 3.7 collaterals/cm(3)), with 97% and 92%, respectively, of the collaterals located within the perfusion territories (intracoronary collaterals). In the normotrophic heart, intracoronary collaterals {median diameter [interquartile range (IQR)]: 91.4 [73.0-115.7] μm} were most prevalent (74%) within the left anterior descending (LAD) territory. Intercoronary collaterals [median diameter (IQR): 94.3 (79.9-107.4) μm] were almost exclusively (99%) found between the LAD and the left circumflex artery (LCX). In the hypertrophied heart, intracoronary collaterals [median diameter (IQR): 101.1 (84.8-126.0) μm] were located within both the LAD (48%) and LCX (46%) territory. Intercoronary collaterals [median diameter (IQR): 97.8 (89.3-111.2) μm] were most prevalent between the LAD-LCX (68%) and LAD-right coronary artery (28%). This study shows that human hearts have abundant coronary collaterals within all flow territories and layers of the heart. The majority of these collaterals are small intracoronary collaterals, which would have remained undetected by clinical imaging techniques., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Thrombospondin-4 (TSP-4) is a multidomain calcium-binding protein that has both intracellular and extracellular functions. As an extracellular matrix protein, it is involved in remodeling processes. Previous work showed that, in the cardiovascular system, TSP-4 expression is induced in the heart in response to experimental pressure overload and infarction injury. Intracellularly, it mediates the endoplasmic reticulum stress response in the heart. In this study, we explored the role of TSP-4 in hypertension. For this purpose, wild-type and TSP-4 knockout (Thbs4(-/-)) mice were treated with angiotensin II (ANG II). Hearts from ANG II-treated Thbs4(-/-) mice showed an exaggerated hypertrophic response. Interestingly, aortas from Thbs4(-/-) mice treated with ANG II showed a high incidence of aneurysms. In resistance arteries, ANG II-treated wild-type mice showed impaired endothelial-dependent relaxation. This was not observed in ANG II-treated Thbs4(-/-) mice or in untreated controls. No differences were found in the passive pressure-diameter curves or stress-strain relationships, although ANG II-treated Thbs4(-/-) mice showed a tendency to be less stiff, associated with thicker diameters of the collagen fibers as revealed by electron microscopy. We conclude that TSP-4 plays a role in hypertension, affecting cardiac hypertrophy, aortic aneurysm formation, as well as endothelial-dependent relaxation in resistance arteries., (Copyright © 2016 the American Physiological Society.)

Transcatheter aortic valve implantation is currently a well-established minimal invasive treatment option for patients with severe aortic valve stenosis. CT Angiography is used for the pre-operative planning and sizing of the prosthesis. To reduce the inconsistency in sizing due to interobserver variability, we introduce and evaluate an automatic aortic root landmarks detection method to determine the sizing parameters. The proposed algorithm detects the sinotubular junction, two coronary ostia, and three valvular hinge points on a segmented aortic root surface. Using these aortic root landmarks, the automated method determines annulus radius, annulus orientation, and distance from annulus plane to right and left coronary ostia. Validation is performed by the comparison with manual measurements of two observers for 40 CTA image datasets. Detection of landmarks showed high accuracy where the mean distance between the automatically detected and reference landmarks was 2.81 ± 2.08 mm, comparable to the interobserver variation of 2.67 ± 2.52 mm. The mean annulus to coronary ostium distance was 16.9 ± 3.3 and 17.1 ± 3.3 mm for the automated and the reference manual measurements, respectively, with a mean paired difference of 1.89 ± 1.71 mm and interobserver mean paired difference of 1.38 ± 1.52 mm. Automated detection of aortic root landmarks enables automated sizing with good agreement with manual measurements, which suggests applicability of the presented method in current clinical practice.

Background: In the absence of a true lymphatic system in the brain parenchyma, alternative clearance pathways for excess fluid and waste products have been proposed. Suggested mechanisms for clearance implicate a role for brain interstitial and cerebrospinal fluids. However, the proposed direction of flow, the anatomical structures involved, and the driving forces are controversial., Methods: To trace the distribution of interstitial and cerebrospinal fluid in the brain, and to identify the anatomical structures involved, we infused a mix of fluorescent tracers with different sizes into the cisterna magna or striatum of mouse brains. We subsequently performed confocal fluorescence imaging of horizontal brain sections and made 3D reconstructions of the mouse brain and vasculature., Results: We observed a distribution pattern of tracers from the parenchyma to the ventricular system, from where tracers mixed with the cerebrospinal fluid, reached the subarachnoid space, and left the brain via the cribriform plate and the nose. Tracers also entered paravascular spaces around arteries both after injection in the cisterna magna and striatum, but this appeared to be of minor importance., Conclusion: These data suggest a bulk flow of interstitial fluid from the striatum towards the adjacent lateral ventricle. Tracers may enter arterial paravascular spaces from two sides, both through bulk flow from the parenchyma and through mixing of CSF in the subarachnoid space. Disturbances in this transport pathway could influence the drainage of amyloid β and other waste products, which may be relevant for the pathophysiology of Alzheimer's disease.

Small arteries are known to develop functional and structural alterations in hypertension. However, the mechanisms of this remodeling are not fully understood. We hypothesized that altered gene expression is associated with the development of hypertension in mesenteric arteries of spontaneously hypertensive rats (SHR). Three sublines of SHR and normotensive Wistar Kyoto rats (WKY) were studied at 6 weeks and 5 months of age. MiRNA and mRNA microarray experiments were performed and analyzed with bioinformatical tools, including Ingenuity Pathway Analysis (IPA). Principal component analysis showed a clear separation in both miRNA and mRNA expression levels between both ages studied, demonstrating strong age-related changes in expression. At the miRNA level, IPA identified differences between SHR and WKY related to metabolic diseases, cellular growth, and proliferation. The mRNAs differentially expressed between SHR and WKY were related to metabolism, cellular movement and proliferation. The most strongly upregulated gene (9.2-fold) was thrombospondin 4 (Thbs4), a protein involved in the endoplasmic reticulum (ER) stress response that activates transcription factor 6α (ATF6α). ATF6α downstream targets were also differentially expressed in SHR vs. WKY. Differential expression of THBS4, the cleaved form of ATF6α, and two of its targets were further confirmed at the protein level by western blot. In summary, these data revealed a number of genes (n = 202) and miRNAs (n = 3) in mesenteric arteries of SHR that had not been related to hypertension previously. The most prominent of these, Thbs4, is related to vascular ER stress that is associated with hypertension.

Endothelial shear stress (ESS) dynamics are a major determinant of atherosclerosis development. The frequently used Poiseuille method to estimate ESS dynamics has important limitations. Therefore, we investigated whether Womersley flow may provide a better alternative for estimation of ESS while requiring equally simple hemodynamic parameters. Common carotid blood flow, centerline velocity, lumen diameter and mean wall thickness (MWT) were measured with 3T-MRI in 45 subjects at three different occasions. Mean ESS and two measures of pulsatility [shear pulsatility index (SPI) and oscillatory shear index (OSI)] were estimated based on Poiseuille and Womersley flow and compared to the more complex velocity gradient modelling method. The association between ESS and MWT was tested with multiple linear regression analysis; interscan reproducibility was assessed using intraclass correlation coefficients (ICC). Mean ESS and pulsatility indices based on Womersley flow (ESSwq β = -0.18, P = 0.04; SPIwq β = 0.24, P = 0.02; OSIwq β = 0.18, P = 0.045), showed equally good correlations with carotid MWT as the velocity gradient method (ESSvg β = -0.23, P = 0.01; SPIvg β = 0.21, P = 0.02; OSIvg β = 0.07, P = 0.47). This in contrast to the Poiseuille flow method that only showed a good correlation for mean ESS (ESSpq β = -0.18, P = 0.04; SPIpq β = 0.14, P = 0.14; OSIpq β = 0.04, P = 0.69). Womersley and Poiseuille methods had high intraclass correlation coefficients indicating good interscan reproducibility (both ICC = 0.84, 95% confidence interval 0.75-0.90). Estimation of ESS dynamics based on Womersley flow modelling is superior to Poiseuille flow modelling and has good interscan reproducibility.

Purpose: To assess the accuracy and precision of a volumetric wall shear stress (WSS) calculation method applied to cine phase contrast magnetic resonance imaging (PC-MRI) data., Materials and Methods: Volumetric WSS vectors were calculated in software phantoms. WSS algorithm parameters were optimized and the influence of spatial resolution and segmentation was evaluated. Subsequently, 2D cine PC-MRI data in the carotid and the aorta at varying spatial resolutions were obtained (n = 2) and compared with the simulations. Finally, volumetric WSS was calculated in 3D cine PC-MRI data of the carotid bifurcation and the aorta (n = 6)., Results: We found that at least 8 voxels across the diameter are required to obtain a WSS accuracy of 5% and a precision of 20% in software phantoms. Systematic WSS quantification errors up to 40% were found in the case of segmentation errors. The in vivo measurements using 2D cine PC-MRI exhibited WSS increase at increasing spatial resolutions, similar to the results in software phantoms. Volumetric WSS vectors were successfully calculated in three healthy carotid bifurcations and aortas., Conclusion: The effects of resolution and segmentation on the accuracy and precision of the WSS algorithm were quantified. We were able to calculate volumetric WSS in the carotid bifurcation and the aorta., (© 2014 Wiley Periodicals, Inc.)

Vasospasm is known to contribute to delayed cerebral ischemia following subarachnoid hemorrhage (SAH). We hypothesized that vasospasm initiates structural changes within the vessel wall, possibly aggravating ischemia and leading to resistance to vasodilator treatment. We therefore investigated the effect of blood on cerebral arteries with respect to contractile activation and vascular remodeling. In vitro experiments on rodent basilar and middle cerebral arteries showed a gradual contraction in response to overnight exposure to blood. After incubation with blood, a clear inward remodeling was found, reducing the caliber of the passive vessel. The transglutaminase inhibitor L682.777 fully prevented this remodeling. Translation of the in vitro findings to an in vivo SAH model was attempted in rats, using both a single prechiasmatic blood injection model and a double cisterna magna injection model, and in mice, using a single prechiasmatic blood injection. However, we found no substantial changes in active or passive biomechanical properties in vivo. We conclude that extravascular blood can induce matrix remodeling in cerebral arteries, which reduces vascular caliber. This remodeling depends on transglutaminase activity. However, the current rodent SAH models do not permit in vivo confirmation of this mechanism., (© 2015 S. Karger AG, Basel.)

Objectives: Spontaneously hypertensive rats (SHR) have been used frequently as a model for human essential hypertension. However, both the SHR and its normotensive control, the Wistar Kyoto rat (WKY), consist of genetically different sublines. We tested the hypothesis that the pathophysiology of vascular remodeling in hypertension differs among rat sublines., Methods and Results: We studied mesenteric resistance arteries of WKY and SHR from three different sources, at 6 weeks and 5 months of age. Sublines of WKY and SHR showed differences in blood pressure, body weight, vascular remodeling, endothelial function, and vessel ultrastructure. Common features in small mesenteric arteries from SHR were an increase in wall thickness, wall-to-lumen ratio, and internal elastic lamina thickness., Conclusions: Endothelial dysfunction, vascular stiffening, and inward remodeling of small mesenteric arteries are not common features of hypertension, but are subline-dependent. Differences in genetic background associate with different types of vascular remodeling in hypertensive rats.

Wall shear stress (WSS) is involved in many pathophysiological processes related to cardiovascular diseases, and knowledge of WSS may provide vital information on disease progression. WSS is generally quantified with computational fluid dynamics (CFD), but can also be calculated using phase contrast MRI (PC-MRI) measurements. In this study, our objectives were to calculate WSS on the entire luminal surface of human carotid arteries using PC-MRI velocities (WSSMRI ) and to compare it with WSS based on CFD (WSSCFD ). Six healthy volunteers were scanned with a 3 T MRI scanner. WSSCFD was calculated using a generalized flow waveform with a mean flow equal to the mean measured flow. WSSMRI was calculated by estimating the velocity gradient along the inward normal of each mesh node on the luminal surface. Furthermore, WSS was calculated for a down-sampled CFD velocity field mimicking the MRI resolution (WSSCFDlowres ). To ensure minimum temporal variation, WSS was analyzed only at diastole. The patterns of WSSCFD and WSSMRI were compared by quantifying the overlap between low, medium and high WSS tertiles. Finally, WSS directions were compared by calculating the angles between the WSSCFD and WSSMRI vectors. WSSMRI magnitude was found to be lower than WSSCFD (0.62 ± 0.18 Pa versus 0.88 ± 0.30 Pa, p < 0.01) but closer to WSSCFDlowres (0.56 ± 0.18 Pa, p < 0.01). WSSMRI patterns matched well with those of WSSCFD. The overlap area was 68.7 ± 4.4% in low and 69.0 ± 8.9% in high WSS tertiles. The angles between WSSMRI and WSSCFD vectors were small in the high WSS tertiles (20.3 ± 8.2°), but larger in the low WSS tertiles (65.6 ± 17.4°). In conclusion, although WSSMRI magnitude was lower than WSSCFD , the spatial WSS patterns at diastole, which are more relevant to the vascular biology, were similar. PC-MRI-based WSS has potential to be used in the clinic to indicate regions of low and high WSS and the direction of WSS, especially in regions of high WSS., (Copyright © 2014 John Wiley & Sons, Ltd.)

Object: While pseudo-continuous arterial spin labeling (pCASL) is a promising imaging technique to visualize cerebral blood flow, it is also (acoustically) very loud during labeling. In this paper, we reduced the labeling loudness on our scanner by increasing the interval between the RF pulses from the literature standard of 1.0 ms. We also propose recommendations to reduce the loudness on scanners of the same type at other sites., Materials and Methods: First, the sound pressure level (SPL) was both simulated and measured as a function of the labeling interval (1.0-1.8 ms) and longitudinal position in the scanner (-10 to +10 cm, relative to isocenter). Subsequently, we selected the labeling interval with the lowest overall SPL for the "SPL-optimized" pCASL sequence. Nine volunteers were scanned to compare raw signal intensity, temporal signal-to-noise ratio (tSNR) and labeling efficiency between the SPL-optimized and the standard PCASL sequence., Results: Sound pressure level measurements on our scanner showed that loudness was reduced by 6.5 dB at the approximate location of the ear by adjusting the labeling interval to 1.4 ms. Furthermore, image quality was not affected, since no significant differences in signal intensity, tSNR and labeling efficiency were observed., Conclusion: By increasing the pCASL labeling interval, acoustic noise in the pCASL sequence was reduced with 6.5 dB, while image quality was preserved.

Objective: Intracerebral hemorrhages (ICHs) are common in patients with cerebral venous thrombosis (CVT). We examined whether small juxtacortical hemorrhages (JCHs) are characteristic for CVT and studied their radiological and pathological properties., Methods: We identified all patients with CVT and an ICH at baseline admitted between 2000 and 2011 (prospectively from July 2006). JCH was defined as a hemorrhage (diameter < 20mm) located in the white matter just below the cortex. To determine the specificity of JCHs for CVT, we examined the frequency of JCHs in a control group of patients of similar age with an ICH not related to CVT., Results: Of 114 patients with CVT, 53 had an ICH. JCHs were present in 14 of the 53 (26%). The remaining 39 had other kinds of hemorrhages. Papilledema was more common among patients with a JCH compared to patients with other types of ICHs (44% vs 9%, p = 0.01). All patients with a JCH except 1 had thrombosis of the superior sagittal sinus, compared to 49% of patients with CVT and other kinds of hemorrhages (p = 0.004). Reanalysis of all imaging data and histopathologic analysis in 1 patient showed that JCHs are located near the U-fibers and that they follow the curvature of the cortex. Among 196 control patients (spontaneous ICH, not caused by CVT), only 3 patients had a JCH. One of these 3 appeared on re-examination of all imaging results to have had CVT., Interpretation: Small nontraumatic JCHs are a characteristic feature of CVT and are rarely encountered in other conditions., (© 2014 American Neurological Association.)

Introduction: Head movement during CT brain perfusion (CTP) acquisition can deteriorate the accuracy of CTP analysis. Most CTP software packages can only correct in-plane movement and are limited to small ranges. The purpose of this study is to validate a novel 3D correction method for head movement during CTP acquisition., Methods: Thirty-five CTP datasets that were classified as defective due to head movement were included in this study. All CTP time frames were registered with non-contrast CT data using a 3D rigid registration method. Location and appearance of ischemic area in summary maps derived from original and registered CTP datasets were qualitative compared with follow-up non-contrast CT. A quality score (QS) of 0 to 3 was used to express the degree of agreement. Furthermore, experts compared the quality of both summary maps and assigned the improvement score (IS) of the CTP analysis, ranging from -2 (much worse) to 2 (much better)., Results: Summary maps generated from corrected CTP significantly agreed better with appearance of infarct on follow-up CT with mean QS 2.3 versus mean QS 1.8 for summary maps from original CTP (P = 0.024). In comparison to original CTP data, correction resulted in a quality improvement with average IS 0.8: 17 % worsened (IS = -2, -1), 20 % remained unchanged (IS = 0), and 63 % improved (IS = +1, +2)., Conclusion: The proposed 3D movement correction improves the summary map quality for CTP datasets with severe head movement.

Organ perfusion is regulated by vasoactivity and structural adaptation of small arteries and arterioles. These resistance vessels are sensitive to pressure, flow and a range of vasoactive stimuli. Several strongly interacting control loops exist. As an example, the myogenic response to a change of pressure influences the endothelial shear stress, thereby altering the contribution of shear-dependent dilation to the vascular tone. In addition, acute responses change the stimulus for structural adaptation and vice versa. Such control loops are able to maintain resistance vessels in a functional and stable state, characterized by regulated wall stress, shear stress, matched active and passive biomechanics and presence of vascular reserve. In this modeling study, four adaptation processes are identified that together with biomechanical properties effectuate such integrated regulation: control of tone, smooth muscle cell length adaptation, eutrophic matrix rearrangement and trophic responses. Their combined action maintains arteries in their optimal state, ready to cope with new challenges, allowing continuous long-term vasoregulation. The exclusion of any of these processes results in a poorly regulated state and in some cases instability of vascular structure.

Purpose: To evaluate wall shear stress (WSS) estimations in an in vitro and in vivo intracranial aneurysm, WSS was estimated from phase contrast magnetic resonance imaging (PC-MRI) and compared with computational fluid dynamics (CFD)., Materials and Methods: First, WSS was estimated using a high-resolution in vitro PC-MRI measurement under steady and pulsatile flow conditions and compared with CFD simulations. Second, WSS was estimated in steady PC-MRI data acquired at different spatial resolutions. Third, WSS estimations in pulsatile in vivo data were compared with CFD. The direction and magnitude of WSS vectors were computed and compared., Results: Quantitative agreement between PC-MRI and CFD-based WSS estimations was moderate for the phantom (Spearman ρ = 0.69). The WSS magnitude derived from PC-MRI data was lower than CFD for both the in vitro and in vivo case. However, there was qualitative agreement between PC-MRI and CFD, i.e. WSS vector direction was similar for both modalities. Circular WSS patterns were found both in vitro and in vivo for PC-MRI and CFD. Increasing PC-MRI resolution increased mean WSS magnitude and uncovered complex WSS patterns., Conclusion: WSS patterns can be estimated based on PC-MRI data in in vitro and in vivo aneurysm geometries. Similar WSS directions as CFD can be discerned., (Copyright © 2013 Wiley Periodicals, Inc.)

Small artery remodeling involves matrix reorganization, but may also encompass changed smooth muscle cell biomechanical properties. Here we study the temporal relationship between such contractile plasticity and matrix remodeling in small rat mesenteric arteries subjected to 1 or 3 days of altered flow or acute interventions on matrix structure; cross-linking by transglutaminase and matrix digestion by elastase. Diameter-tension relations were made in the passive state and upon full activation (125 mM K+ and 10⁻⁵ M norepinephrine). In low flow (LF), inward matrix remodeling occurred after 1 day, when the distended diameter at full dilation (D₁₀₀) was reduced from 351±15μm to 299±14μm (SEM, n=8, p<0.05). The optimal diameter for force development (D(opt)) was reduced after 3 days, from 291±10μm to 247±5μm (LF, p<0.05). As a result, a mismatch of D(opt)/D₁₀₀ existed after 1 day of LF, which normalized after 3 days. Dynamics of contraction were studied following quick isometric release by 0.2∙D₁₀₀; tension recovery was faster in anatomically smaller vessels following normal flow. This association was partly lost after 1 day of LF, while after 3 days the vessels became not only smaller but also faster, re-establishing this association. High flow vessels demonstrated similar contractile plasticity. Active diameter-tension relations at low distension did not change following transglutaminase or elastase. However, at high distension, any alteration in passive tension coincided with an opposite change in active tension. These data demonstrate an intrinsic interaction between passive and active biomechanics that occurs instantaneously during matrix remodeling at high distensions while contractile plasticity lags matrix remodeling after flow interventions., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Adhesion G protein-coupled receptors (aGPCRs) are two-subunit molecules, consisting of an adhesive extracellular α subunit that couples noncovalently to a seven-transmembrane β subunit. The cooperation between the two subunits and the effect of endogenous ligands on the functioning of aGPCRs is poorly understood. In this study, we investigated the interaction between the pan-leukocyte aGPCR CD97 and its ligand CD55. We found that leukocytes from CD55-deficient mice express significantly increased levels of cell surface CD97 that normalized after transfer into wild-type mice because of contact with CD55 on both leukocytes and stromal cells. Downregulation of both CD97 subunits occurred within minutes after first contact with CD55 in vivo, which correlated with an increase in plasma levels of soluble CD97. In vitro, downregulation of CD97 on CD55-deficient leukocytes cocultured with wild-type blood cells was strictly dependent on shear stress. In vivo, CD55-mediated downregulation of CD97 required an intact circulation and was not observed on cells that lack contact with the blood stream, such as microglia. Notably, de novo ligation of CD97 did not activate signaling molecules constitutively engaged by CD97 in cancer cells, such as ERK and protein kinase B/Akt. We conclude that CD55 downregulates CD97 surface expression on circulating leukocytes by a process that requires physical forces, but based on current evidence does not induce receptor signaling. This regulation can restrict CD97-CD55-mediated cell adhesion to tissue sites.

Purpose: To determine the clinical feasibility of arterial spin labeling (ASL) on a 1T open bore scanner., Materials and Methods: First, the optimal postlabeling delay (PLD) at 1T was determined (n = 5), with and without vascular crushing. Second, the effect of different labeling approaches (pseudo-continuous ASL [pCASL] vs. pulsed ASL [PASL]), background suppression (BSup) and readout options (GRASE vs. EPI) was investigated (n = 9). Each effect was quantified by calculating the signal-to-noise ratio (SNR), convergence, and number of significant gray matter (GM) voxels in the ASL images. Finally, an example of an obese volunteer who could not have been scanned in a cylindrical scanner is presented., Results: The optimal PLDs were found to be 1300 msec for pCASL with and without vascular crushing. pCASL labeling outperformed PASL labeling in terms of convergence, anatomical correspondence between GM and perfusion maps, and SNR (P < 0.05). BSup appeared to have no additional value on the convergence, anatomical GM correspondence, and SNR (P > 0.05). EPI readout yielded a slightly better convergence, while the SNR of the GRASE readout was higher (P < 0.05)., Conclusion: ASL on 1T is clinically feasible using state-of-the-art sequences that were primarily developed for higher field strengths., (Copyright © 2012 Wiley Periodicals, Inc.)

Inward remodeling of small arteries occurs after prolonged vasoconstriction, low blood flow, and in several models of hypertension. The cross-linking enzyme, transglutaminases 2 (TG2), is able to induce inward remodeling and stiffening of arteries. The activity of TG2 is dependent on its conformation, which can be open or closed, and on its redox state. Several factors have been shown to be involved in modulating TG2 activity, including Ca(2+) and GTP/GDP concentrations, as well as the redox state of the environment. This review introduces the hypothesis that mechanical force could be involved in regulating the activity of TG2 during inward remodeling by promoting its open and reduced active state. Several aspects of TG2, such as its structure and localization, are assessed in order to provide arguments that support the hypothesis. We conclude that a direct activation of TG2 by mechanical force exerted by smooth muscle cells may explain the link between smooth muscle activation and inward remodeling, as observed in several physiological and pathological conditions., (Copyright © 2013 S. Karger AG, Basel.)

Small artery remodeling may involve a shift in the diameter-dependent force generating capacity of smooth muscle cells (SMC). We tested to what extent and under which conditions such contractile plasticity occurs. Rat mesenteric arteries were mounted on isometric myographs. Active diameter-tension relations were determined after application of several stimuli for 16 or 40 h at 40 or 110% of the passive diameter at 100 mm Hg. At 40%, 16-hour incubation with endothelin-1 (ET-1) but not U46619 shifted force capacity towards smaller diameters. Inflammatory cytokines (TNF-α, IL-1β, IFN-γ), TGF-β or serum neither induced such shift nor augmented the effect of ET-1. The ET-1-mediated change was not affected by superoxide dismutase and catalase. Inward matrix remodeling in the presence of ET-1 was slower, occurring after 40 h. Arteries maintained at 110% showed a shift of force capacity to larger diameters, which was prevented by ET-1 but not by U46619. In the active but not the passive state, SMC had altered nuclear lengths after incubation at 40%. These data demonstrate contractile plasticity in small arteries, where chronic strain is an outward drive and specifically ET-1 an inward drive, acting through mechanisms that do not seem to relate to oxidative stress, inflammatory pathways or major reorganization of the SMC., (© 2013 S. Karger AG, Basel.)

Objective: Atherosclerosis preferentially develops at sites of disturbed blood flow. We tested the hypothesis that transglutaminase activity plays a role in plaque development at these locations., Methods and Results: Exposure of endothelial cells to steady flow (7 dynes/cm(2)) was associated with relatively low transglutaminase activity, whereas under low oscillatory flow (1.3 ± 2.6 dynes/cm(2)) endothelial cells showed a >4-fold higher level of transglutaminase activity. Under oscillatory flow, transglutaminase activity increased the expression of the chemokine MCP-1 (CCL2). In vivo, oscillatory flow was induced by placement of a tapered perivascular cast around the carotid artery of type 2 transglutaminase (TGM2) knockout mice and WT counterparts. After 2 days, significantly less monocytes adhered to the endothelium in TGM2 knockout mice as compared to WT. In a more chronic setting, ApoE knockout mice that were equipped with the flow-modifying cast developed lesions proximal to the cast (low shear stress), and distal to the cast (oscillatory shear stress). Inhibition of transglutaminase induced a marked reduction in macrophage and fat content in distal lesions only. In addition, lesion size was increased in this area, which was attributed to an increase in smooth muscle content., Conclusion: Oscillatory shear stress increases endothelial transglutaminase activity. In turn, transglutaminase activity affects the expression of MCP-1 in vitro and monocyte recruitment in vivo. In a mouse model of atherosclerosis, transglutaminase activity has a major effect on plaque composition under oscillatory shear stress., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Transglutaminase 2 (TG2) is a pleiotropic enzyme involved in both intra- and extracellular processes. In the extracellular matrix, TG2 stabilizes the matrix by both covalent cross-linking and disulfide isomerase activity. These functions become especially apparent during matrix remodeling as seen in wound healing, tumor development and vascular remodeling. However, TG2 lacks the signal sequence for a classical secretory mechanism, and the cellular mechanism of TG2 secretion is currently unknown. We developed a green fluorescent TG2 fusion protein to study the hypothesis that TG2 is secreted via microparticles. Characterization of TG2/eGFP, using HEK/293T cells with a low endogenous TG2 expression, showed that cross-linking activity and fibronectin binding were unaffected. Transfection of TG2/eGFP into smooth muscle cells resulted in the formation of microparticles (MPs) enriched in TG2, as detected both by immunofluorescent microscopy and flow cytometry. The fraction of TG2-positive MPs was significantly lower for cross-linking-deficient mutants of TG2, implicating a functional role for TG2 in the formation of MPs. In conclusion, the current data suggest that TG2 is secreted from the cell via microparticles through a process regulated by TG2 cross-linking.

Blood vessel structure and calibre are not static. Rather, vessels remodel continuously in response to their biomechanical environment. Vascular calibre is dictated by the amount, composition and organization of the elastic extracellular matrix. In addition, the amount and organization of contractile smooth muscle cell (SMC) also need to be regulated. The SMCs are organized such that maximum contractile force generally occurs at diameters slightly below the diameter at full dilation and physiological pressure. Thus, in a remodelling vessel, not only the matrix but also the SMCs need to undergo structural adaptation. Surprisingly little is known on the adaptation of SMC contractile properties in the vasculature. The purpose of this review is to explore this SMC plasticity in the context of vascular remodelling. While not much work on this has been carried out on blood vessels, SMC plasticity is more extensively studied on other hollow structures such as airway and bladder. We therefore include studies on bladder and airway SMCs because of their possible relevance for vascular SMC behaviour. Here, plasticity is thought to form an adaptation allowing maintained function despite large volume changes. In blood vessels, the general match of active and passive diameter-tension relations suggests that SMC plasticity is part of normal vascular physiological adaptation. Vascular SMCs display similar processes and forms of adaptation as seen in nonvascular SMCs. This may become particularly relevant under strong vasoconstriction, when inward cytoskeletal adaptation possibly prevents immediate full dilation. This may contribute to structural inward remodelling as seen in hypertension and flow reduction., (© 2011 The Authors. Basic & Clinical Pharmacology & Toxicology © 2011 Nordic Pharmacological Society.)

Straightforward quantification of variations of flow patterns within aneurysms fails to accurately describe flow patterns of interest. We applied a multiscale decomposition of the flow in well-defined patterns to detect and quantify flow patterns in an aneurysm phantom that was studied with three different modalities: MRI, computational fluid dynamics, and particle image velocimetry. The method intuitively visualizes main patterns such as locally uniform flow, in- and outflow, and vortices. It is shown that this method is a valuable tool to quantitatively compare scale-dependent complex flow patterns in aneurysms.

While inward remodeling of small arteries in response to low blood flow, hypertension, and chronic vasoconstriction depends on type 2 transglutaminase (TG2), the mechanisms of action have remained unresolved. We studied the regulation of TG2 activity, its (sub) cellular localization, substrates, and its specific mode of action during small artery inward remodeling. We found that inward remodeling of isolated mouse mesenteric arteries by exogenous TG2 required the presence of a reducing agent. The effect of TG2 depended on its cross-linking activity, as indicated by the lack of effect of mutant TG2. The cell-permeable reducing agent DTT, but not the cell-impermeable reducing agent TCEP, induced translocation of endogenous TG2 and high membrane-bound transglutaminase activity. This coincided with inward remodeling, characterized by a stiffening of the artery. The remodeling could be inhibited by a TG2 inhibitor and by the nitric oxide donor, SNAP. Using a pull-down assay and mass spectrometry, 21 proteins were identified as TG2 cross-linking substrates, including fibronectin, collagen and nidogen. Inward remodeling induced by low blood flow was associated with the upregulation of several anti-oxidant proteins, notably glutathione-S-transferase, and selenoprotein P. In conclusion, these results show that a reduced state induces smooth muscle membrane-bound TG2 activity. Inward remodeling results from the cross-linking of vicinal matrix proteins, causing a stiffening of the arterial wall.