Arterial stiffening is influenced by the organization of focal adhesions in vascular smooth muscle cells (VSMCs). We investigated the contribution of αv integrins to both arterial wall stiffness (Young’s modulus measured by echography) and VSMC stiffness (assessed by atomic force microscopy). Mice with VSMC-specific deletion of αv integrins (αvSMKO) were compared with controls at baseline and following angiotensin II infusion. Unstimulated cultured αv-deficient (αv-KD) VSMCs exhibited higher stiffness than controls, with a further increase after angiotensin II. To interpret AFM measurements performed at shallow indentation depths, we developed a computational model of VSMC nanoindentation. Simulations showed that higher apparent Young’s moduli at shallow indentation fall within the experimental range of αv-KD cells. These cells also displayed enhanced actin polymerization, further amplified by angiotensin II through the formation of cortical F-actin. In vivo, arterial pressure and wall elastic modulus were similar between αvSMKO and control mice at baseline and after angiotensin II, despite αvSMKO mice exhibiting lower elastin and higher collagen content under angiotensin II. Together, these findings indicate that the comparable increase in arterial stiffness observed in αvSMKO mice under angiotensin II is driven primarily by elevated VSMC stiffness resulting from cortical actin redistribution, which outweighs extracellular matrix changes.

Abstract Aims αv integrins are implicated in fibrosis in a number of organs through their ability to activate TGF-β. However their role in vascular fibrosis and collagen accumulation is only partially understood. Here we have used αv conditional knockout mice and cell lines to determine how αv contributes to vascular smooth muscle cell (VSMC) function in vascular fibrosis and the role of TGF-β in that process. Methods and results Angiotensin II (Ang II) treatment causes upregulation of αv and β3 expression in the vessel wall, associated with increased collagen deposition. We found that deletion of αv integrin subunit from VSMCs (αvSMKO) protected mice against angiotensin II-induced collagen production and assembly. Transcriptomic analysis of the vessel wall in αvSMKO mice and controls identified a significant reduction in expression of fibrosis and related genes in αvSMKO mice. In contrast, αvSMKO mice showed prolonged expression of CD109, which is known to affect TGF-β signalling. Using cultured mouse and human VSMCs, we showed that overexpression of CD109 phenocopied knockdown of αv integrin, attenuating collagen expression, TGF-β activation, and Smad2/3 signalling in response to angiotensin II or TGF-β stimulation. CD109 and TGF-β receptor were internalized in early endosomes. Conclusion We identify a role for VSMC αv integrin in vascular fibrosis and show that αv acts in concert with CD109 to regulate TGF-β signalling.

Single-beam acoustic tweezers have recently been demonstrated to be capable of selective three-dimensional trapping. This new contactless manipulation modality has great potential for many scientific applications. Its development as a scientific tool requires precise calibration of its radiation force, specifically its axial component. The lack of calibration for this force is mainly due to its weak magnitude compared to competing effects such as weight. We investigate an experimental method for the calibration of the axial stiffness of the radiation force by observing the axial oscillations of a trapped bead in a microgravity environment. The stiffness exhibits a linear relationship with the acoustic intensity and is of the mN/m order. Then, a predictive model, loaded with the experimental acoustic field, is compared to the measured stiffness with very good agreement, within a single amplitude coefficient. This study paves the way for the development of calibrated acoustic tweezers.

This review seeks to provide an update of the mechanisms of vascular cell senescence, from newly identified molecules to arterial ageing phenotypes, and finally to present a computational approach to connect these selected proteins in biological networks. We will discuss current key signalling and gene expression pathways by which these focus proteins and networks drive normal and accelerated vascular ageing. We also review the possibility that senolytic drugs, designed to restore normal cell differentiation and function, could effectively treat multiple age-related vascular diseases. Finally, we discuss how cell senescence is both a cause and a consequence of vascular ageing because of the possible feedback controls between identified networks.

This work is part of a study dealing with city-dwellers' quality of life. Noise is known to be an important factor influencing the quality of life. In order to diagnose it properly, we propose a noise monitoring system of urban areas. It is based on the use of large microphone arrays in order to extract the radiated sound field from each passing-by vehicle in typical urban scenes. A machine learning algorithm is trained so as to classify these extracted signals in clusters combining both the vehicle type and the driving conditions. This system makes it possible to monitor the evolution of the noise levels for each cluster. The proposed system was first tested on passing-by isolated vehicles measurements and then implemented in a real street in Paris (France).

Large microphone arrays are an efficient means for source localization thanks to a wide aperture and a great number of sensors. When such arrays are deployed in situ, accurate geometric calibration becomes essential to obtain the microphone positions. In free field, the classic procedures rely on measured Times of Arrival (TOA) or Time Differences of Arrival (TDOA) between the microphones and several controlled sources. However, free field model mismatches, such as reflectors, generate outliers which severely deteriorate the positioning accuracy. This paper introduces a unified framework for robust calibration using TOA or TDOA by exploiting an outlier-aware noise model. Thanks to the largeness of the array, the existing outliers are sparse and can be identified by a Lasso regression. From this, three iterative robust solvers are proposed: (i) for TOA by Robust Multi Dimensional Unfolding, a particular variation of Robust Multi Dimensional Scaling, (ii) for TDOA by data predenoising based on sparse and low-rank matrix decomposition, and (iii) for TDOA by jointly identifying the outliers and the geometry. The relevance of outlier-aware approaches is asserted by numerical and experimental tests. Compared with the baseline least-square approaches, the proposed robust solvers significantly improve the positioning accuracy in a free field mismatched by reflectors.

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Background: The metabolic syndrome (MetS) and aging are associated with modifications in blood coagulation factors, vascular inflammation, and increased risk of thrombosis. Objectives: Our aim was to determine concomitant changes in thrombin generation in the blood compartment and at the surface of vascular smooth muscle cells (VSMCs) and its interplay with adipokines, free fatty acids (FFA), and metalloproteinases (MMPs) in obese Zucker rats that share features of the human MetS. Methods: Obese and age-matched lean Zucker rats were compared at 25 and 80 weeks of age. Thrombin generation was assessed by calibrated automated thrombography (CAT). Results: Endogenous thrombin potential (ETP) was increased in obese rats independent of platelets and age. Clot half-lysis time was delayed with obesity and age. Interleukin (IL)-1β and IL-13 were increased with obesity and age respectively. Addition of exogenous fibrinogen, leptin, linoleic, or palmitic acid increased thrombin generation in plasma whereas adiponectin had an opposite effect. ETP was increased at the surface of VSMCs from obese rats and addition of exogenous palmitic acid further enhanced ETP values. Gelatinase activity was increased in aorta at both ages in obese rats and MMP-2 activity was increased in VSMCs from obese rats. Conclusions: Our study demonstrated in MetS an early prothrombotic phenotype of the blood compartment reinforced by procoagulant properties of dedifferentiated and inflammatory VSMCs. Mechanisms involved (1) increased fibrinogen and impaired fibrinolysis and (2) increased saturated fatty acids responsible for additive procoagulant effects. Whether specifically targeting this hypercoagulability using direct thrombin inhibitors would improve outcome in MetS is worth investigating.

This editorial refers to ‘Inhibition of SRF/myocardin reduces aortic stiffness by targeting vascular smooth muscle cell stiffening in hypertension’ by Zhou et al ., pp. 171–182. Large artery stiffening is a main risk factor for chronic diseases including aging-related hypertension, heart failure, renal disease, stroke and dementia, which represent major causes of morbidity and mortality. Arterial stiffening is caused primarily by excessive fibrosis due to exuberant accumulation of collagen. More recently the mechanisms of arterial stiffness in hypertension were extended to proteins regulating vascular smooth muscle cell (VSMC) plasticity and cellular stiffness.1 Despite earlier studies showing involvement of the cytoskeletal network, there is a gap in our knowledge regarding underlying mechanisms that initiate cellular stiffness and sustain this process. In this issue Zhou et al 2 provide evidence that VSMC stiffness assessed by atomic force microscopy (AFM) is increased in spontaneously hypertensive rats (SHR) at the level of large arteries but not in small arteries. RNA and protein expression of serum response factor (SFR)/myocardin as well as their downstream targets caldesmon, calponin, and RhoA are increased in thoracic aorta from SHR compared with arteries from Wistar-Kyoto rats (WKY) at 4 months of age. The hypertension induced-increase in SRF represents one of the main mechanisms, as supported by normalization of cellular stiffness measurements in the presence of an inhibitor of SRF activity in both in vitro and in vivo conditions. Normalization of cellular stiffness is paralleled by an improvement of aortic elasticity preceding the reduction of blood pressure in SHR. This finding suggests that …

This paper presents a complete strategy for the geometry estimation of large microphone arrays of arbitrary shape. Largeness is intended here in both number of microphones (hundreds) and size (few meters). Such arrays can be used for various applications in open or confined spaces like acoustical imaging, source identification, or speech processing. For so large array systems, measuring the geometry by hand is impractical. Therefore a blind passive method is proposed. It is based on the analysis of the background acoustic noise, supposed to be a diffuse field. The proposed strategy is a two-step process. First the pairwise microphone distances are identified by matching their measured coherence function to the one predicted by the diffuse field theory. Second, a robust multidimensional scaling (MDS) algorithm is adapted and implemented. It takes advantage of local characteristics to reduce the set of distances and infer the geometry of the array. This work is an extension of previous studies, and it overcomes unsolved drawbacks. In particular it deals efficiently with the outliers known to ruin standard MDS algorithms. Experimental proofs of this ability are presented by treating the case of two arrays. They show that the proposed improvements manage large spatial arrays.

A number of key signaling pathways are highly relevant to early vascular aging. These are overviewed here to provide a context for understanding the initiation and progression of hypertension, arteriosclerosis, and atherosclerosis. The elements of the pathways include cytoskeletal and contractile proteins, extracellular matrix proteins, and inflammatory processes. Activation of these pathways leads to a proinflammatory state and tissue oxidative stress, which in turn drives age-associated arterial stiffening processes. A progressive decrease in plasticity and increase in reprogramming potential of vascular smooth muscle cells contributes to the clinical phenotype of aging associated with isolated systolic hypertension, heart failure, diabetes, and stroke.

Noise pollution investigation takes advantage of two common methods of diagnosis: measurement using a Sound Level Meter and acoustical imaging. The former enables a detailed analysis of the surrounding noise spectrum whereas the latter is rather used for source localization. Both approaches complete each other, and merging them into a unique system, working in realtime, would offer new possibilities of dynamic diagnosis. This paper describes the design of a complete system for this purpose: imaging in realtime the acoustic field at different octave bands, with a convenient device. The acoustic field is sampled in time and space using an array of MEMS microphones. This recent technology enables a compact and fully digital design of the system. However, performing realtime imaging with resource-intensive algorithm on a large amount of measured data confronts with a technical challenge. This is overcome by executing the whole process on a Graphic Processing Unit, which has recently become an attractive device for parallel computing.

Objective-The hypothesis that hypertension induces a hypercoagulable state arises from the complications associated with hypertension: stroke and myocardial infarction. Here, we determine whether hypertension causes changes in the thrombin-generating capacity of the vascular wall.Approach and Results-We used spontaneously hypertensive rats (SHR) compared with Wistar rats. The addition of thoracic aortic rings of SHR to a Wistar or SHR plasma pool resulted in a greater increase in thrombin generation compared with equivalent rings from Wistar. This increase occurred in 12-but not 5-week-old rats and was prevented by an angiotensin II-converting enzyme inhibitor, indicating that established hypertension is required to induce increased thrombin generation within the vessel wall. Whereas no difference was observed for endothelial cells, thrombin formation was higher on aortic smooth muscle cells (SMCs) from SHR than on those from Wistar. Exposure of negatively charged phospholipids was higher on SHR than on Wistar rings, as well as on cultured SMCs. Tissue factor activity was higher in SHR SMCs. Twelve-week-old SHR exhibited accelerated FeCl3-induced thrombus formation in carotid arteries, and the resulting occlusive thrombi were disaggregated by blockade of glycoprotein Iba-von Willebrand factor interactions. SHR SMCs were more sensitive to thrombin-induced proliferation than Wistar SMCs. This effect was totally abolished by a protease-activated receptor 1 inhibitor.Conclusions-The prothrombotic phenotype of the SHR vessel wall was due to the ability of SMCs to support greater thrombin generation and resulted in accelerated occlusive thrombus formation after arterial injury, which was sensitive to glycoprotein Iba-von Willebrand factor inhibitors.

Background— We assess the contribution of common and rare putatively functional genetic variants (most of them coding) present on the Illumina exome Beadchip to the variability of plasma lipids and stiffness of the common carotid artery. Methods and Results— Measurements were obtained from 2283 men and 1398 women, and after filtering and exclusion of monomorphic variants, 32 827 common (minor allele frequency >0.01) and 68 770 rare variants were analyzed. A large fraction of the heritability of plasma lipids is attributable to variants present on the array, especially for triglycerides (fraction of variance attributable to measured genotypes: V (G)/ V p =31.4%, P <3.1×10 –11 ) and high-density lipoprotein cholesterol ( V (G)/ V p =26.4%, P <4.2×10 –12 ). Plasma lipids were associated with common variants located in known candidate genes, but no implication of rare variants could be established. Gene sets for plasma lipids, blood pressure, and coronary artery disease were defined on the basis of recent meta-analyses of genome-wide association studies. We observed a strong association between the plasma lipids gene set and plasma lipid variables, but none of the 3 genome-wide association studies gene sets was associated with the carotid parameters. Significant V (G)/ V p ratios were observed for external (14.5%, P <2.7×10 –5 ) and internal diameter (13.4%, P <4.3×10 –4 ), stiffness (12.5%, P <8.0×10 –4 ), intima-media thickness (10.6%, P <7.9×10 –4 ), and wall cross-sectional area (13.2%, P <2.4×10 –5 ). A significant association was observed between the common rs2903692 polymorphism of the CLEC16A gene and the internal diameter ( P <4.3×10 –7 ). Conclusions— These results suggest an involvement of CLEC16A , a gene that has been reported to be associated with immune disorders, in the modulation of carotid vasodilatation.

Rationale: Sinoaortic denervated (SAD) and chemically sympathectomized (SNX) rats are characterized by a decrease in arterial distensibility without hypertension and would, thus, be relevant for analyzing arterial wall stiffening independently of blood pressure level. The fibronectin network, which plays a pivotal role in cell–matrix interactions, is a major determinant of arterial stiffness. We hypothesized that in SAD and SNX rats, arterial stiffness is increased, due to alterations of cell–matrix anchoring leading to spatial reorganization of the extracellular matrix. Methods: The intrinsic elastic properties of the arterial wall were evaluated in vivo by the relationship between incremental elastic modulus determined by echotracking and circumferential wall stress. The changes of cell-extracellular matrix links in the abdominal aorta were evaluated by studying fibronectin, vascular integrin receptors, and ultrastructural features of the aorta by immunochemistry. Results: In both experimental conditions, wall stiffness increased, associated with different modifications of cell-extracellular matrix adhesion. In SAD rats, increased media cross-sectional area was coupled with an increase of muscle cell attachments to its extracellular matrix via fibronectin and its α5-β1 integrin. In SNX rats, reduced media cross-sectional area was associated with upregulation of αv-β3 integrin and more extensive connections between dense bands and elastic fibers despite the disruption of the elastic lamellae. Conclusion: In aorta of SNX and SAD rats, a similar arterial stiffness is associated to different structural alterations. An increase in αvβ3 or α5β1 integrins together with the already reported increase in the proportion of less distensible (collagen) to more distensible (elastin) components in both models contributes to remodeling and stiffening of the abdominal aorta.

RATIONALE:Sinoaortic denervated (SAD) and chemically sympathectomized (SNX) rats are characterized by a decrease in arterial distensibility without hypertension and would, thus, be relevant for analyzing arterial wall stiffening independently of blood pressure level. The fibronectin network, which plays a pivotal role in cell-matrix interactions, is a major determinant of arterial stiffness. We hypothesized that in SAD and SNX rats, arterial stiffness is increased, due to alterations of cell-matrix anchoring leading to spatial reorganization of the extracellular matrix. METHODS:The intrinsic elastic properties of the arterial wall were evaluated in vivo by the relationship between incremental elastic modulus determined by echotracking and circumferential wall stress. The changes of cell-extracellular matrix links in the abdominal aorta were evaluated by studying fibronectin, vascular integrin receptors, and ultrastructural features of the aorta by immunochemistry. RESULTS:In both experimental conditions, wall stiffness increased, associated with different modifications of cell-extracellular matrix adhesion. In SAD rats, increased media cross-sectional area was coupled with an increase of muscle cell attachments to its extracellular matrix via fibronectin and its α5-β1 integrin. In SNX rats, reduced media cross-sectional area was associated with upregulation of αv-β3 integrin and more extensive connections between dense bands and elastic fibers despite the disruption of the elastic lamellae. CONCLUSION:In aorta of SNX and SAD rats, a similar arterial stiffness is associated to different structural alterations. An increase in αvβ3 or α5β1 integrins together with the already reported increase in the proportion of less distensible (collagen) to more distensible (elastin) components in both models contributes to remodeling and stiffening of the abdominal aorta.

Arterial stiffness is recognized as a risk factor for many cardiovascular diseases. Aldosterone via its binding to and activation of the mineralocorticoid receptors (MRs) is a main regulator of blood pressure by controlling renal sodium reabsorption. Although both clinical and experimental data indicate that MR activation by aldosterone is involved in arterial stiffening, the molecular mechanism is not known. In addition to the kidney, MR is expressed in both endothelial and vascular smooth muscle cells (VSMCs), but the specific contribution of the VSMC MR to aldosterone-induced vascular stiffness remains to be explored. To address this question, we generated a mouse model with conditional inactivation of the MR in VSMC (MR SMKO ). MR SMKO mice show no alteration in renal sodium handling or vascular structure, but they have decreased blood pressure when compared with control littermate mice. In vivo at baseline, large vessels of mutant mice presented with normal elastic properties, whereas carotids displayed a smaller diameter when compared with those of the control group. As expected after aldosterone/salt challenge, the arterial stiffness increased in control mice; however, it remained unchanged in MR SMKO mice, without significant modification in vascular collagen/elastin ratio. Instead, we found that the fibronectin/α5-subunit integrin ratio is profoundly altered in MR SMKO mice because the induction of α5 expression by aldosterone/salt challenge is prevented in mice lacking VSMC MR. Altogether, our data reveal in the aldosterone/salt hypertension model that MR activation specifically in VSMC leads to the arterial stiffening by modulation of cell-matrix attachment proteins independent of major vascular structural changes.

Although hypertension contributes significantly to worsen cardiovascular risk, blood pressure increment in subjects with heart failure is paradoxically associated with lower risk. The objective was to determine whether pulse pressure and pulse wave velocity (PWV) remain prognostic markers, independent of treatment in heart failure with reduced left ventricular function. The investigation involved 6632 patients of the Eplerenone Post–Acute Myocardial Infarction Heart Failure Efficacy and Survival Study. All subjects had acute myocardial infarction with left ventricular ejection fraction <40% and signs/symptoms of heart failure. Carotid-femoral PWV was measured in a subpopulation of 306 subjects. In the overall population, baseline mean arterial pressure <90 mm Hg was associated with higher all-cause death (hazard ratio, 1.14 [95% confidence interval, 1.00–1.30]; P <0.05), whereas higher left ventricular ejection fraction or pulse pressure was associated with lower rates of all-cause death, cardiovascular death/hospitalization, and cardiovascular death. In the subpopulation, increased baseline PWV was associated with worse outcomes (all-cause death: 1.16 [1.03–1.30]; P <0.05 and cardiovascular deaths: 1.16 [1.03–1.31]; P <0.05), independent of age and left ventricular ejection fraction. Using multiple regression analysis, systolic blood pressure and age were the main independent factors positively associated with pulse pressure or PWV, both in the entire population or in the PWV substudy. In heart failure and low ejection fraction, our results suggest that pulse pressure, being negatively associated with outcome, is more dependent on left ventricular function and thereby no longer a marker of aortic elasticity. In contrast, increased aortic stiffness, assessed by PWV, contributes significantly to cardiovascular death.

The hypothesis that hypertension may confer a hypercoagulable state arises from the main complications associated with hypertension, stroke and myocardial infarction. Our objective was to determine whether spontaneous hypertension confers changes in the coagulation proteins and the thrombin generating capacity in blood and the vascular wall. We used the model of spontaneously hypertensive rats (SHR) compared with Wistar rats. Thrombin generation was lower in platelet-rich plasma and platelet-free plasma from SHR compared to Wistar. This was related to lower tissue factor (TF) and prothrombin as well as higher TFPI levels in SHR plasma. In contrast, the addition of thoracic aorta rings of SHR to a Wistar plasma pool resulted in a higher increase in thrombin generation compared to the addition of equivalent rings from Wistar. Whereas no difference was observed for endothelial cells, thrombin formation was higher at the surface of cultured SHR aortic SMCs than from Wistar. Exposure of negatively-charged phospholipids was higher on SHR than on Wistar rings as well as on SMCs. TF and TFPI activities were higher in SHR SMCs. These results show opposite thrombin generating capacity of plasma and vessel walls in SHR compared to Wistar. The higher prothrombotic phenotype of the SHR vessel wall was due to the ability of SMCs to support thrombin generation. These findings suggest that the hypertension-induced membrane phospholipid reorganization and synthesis of procoagulant molecules in SMCs provide substrates for increased thrombin formation within the vessel wall.