Stress Load and Ascending Aortic Aneurysms: An Observational, Longitudinal, Single-Center Study Using Computational Fluid Dynamics.

Ascending aortic aneurysm (AAoA) is a silent disease with high mortality; however, the factors associated with a worse prognosis are not completely understood. The objective of this observational, longitudinal, single-center study was to identify the hemodynamic patterns and their influence on AAoA growth using computational fluid dynamics (CFD), focusing on the effects of geometrical variations on aortic hemodynamics. Personalized anatomic models were obtained from angiotomography scans of 30 patients in two different years (with intervals of one to three years between them), of which 16 (53%) showed aneurysm growth (defined as an increase in the ascending aorta volume by 5% or more). Numerically determined velocity and pressure fields were compared with the outcome of aneurysm growth. Through a statistical analysis, hemodynamic characteristics were found to be associated with aneurysm growth: average and maximum high pressure (superior to 100 Pa); average and maximum high wall shear stress (superior to 7 Pa) combined with high pressure (>100 Pa); and stress load over time (maximum pressure multiplied by the time interval between the exams). This study provides insights into a worse prognosis of this serious disease and may collaborate for the expansion of knowledge about mechanobiology in the progression of AAoA.

Epigenetic changes in shear-stressed endothelial cells.

Epigenetic changes, particularly histone compaction modifications, have emerged as critical regulators in the epigenetic pathway driving endothelial cell phenotype under constant exposure to laminar forces induced by blood flow. However, the underlying epigenetic mechanisms governing endothelial cell behavior in this context remain poorly understood. To address this knowledge gap, we conducted in vitro experiments using human umbilical vein endothelial cells subjected to various tensional forces simulating pathophysiological blood flow shear stress conditions, ranging from normotensive to hypertensive forces. Our study uncovers a noteworthy observation wherein endothelial cells exposed to high shear stress demonstrate a decrease in the epigenetic marks H3K4ac and H3K27ac, accompanied by significant alterations in the levels of HDAC (histone deacetylase) proteins. Moreover, we demonstrate a negative regulatory effect of increased shear stress on HOXA13 gene expression and a concomitant increase in the expression of the long noncoding RNA, HOTTIP, suggesting a direct association with the suppression of HOXA13. Collectively, these findings represent the first evidence of the role of histone-related epigenetic modifications in modulating chromatin compaction during mechanosignaling of endothelial cells in response to elevated shear stress forces. Additionally, our results highlight the importance of understanding the physiological role of HOXA13 in vascular biology and hypertensive patients, emphasizing the potential for developing small molecules to modulate its activity. These findings warrant further preclinical investigations and open new avenues for therapeutic interventions targeting epigenetic mechanisms in hypertensive conditions.

Modeling mass removal and sediment deposition in stormwater ponds using floating treatment islands: a computational approach.

Floating treatment islands (FTIs) offer effective solutions for stormwater management, providing flood attenuation and pollutant removal capabilities. However, there remains a knowledge gap concerning their performance, specifically in terms of pollutant removal and sediment deposition. To address this gap, the present study employs computational fluid dynamics (CFD) modeling to investigate the intricate interactions within FTI systems. Various FTI configurations are analyzed, considering mass removal through FTIs and sediment deposition, the first time where these two processes were considered together in a CFD environment. The findings demonstrate that FTIs have a significant influence on flow patterns and mass removal. Notably, FTIs enhance mass removal compared to the control case, with larger sediment particles exhibiting higher removal rates. The correlation between the short-circuit index and sedimentation in FTI ponds highlights the potential of FTIs as indicators of treatment efficiency. Furthermore, the study focuses on mass removal exclusively through the FTI root zones. The positioning of FTIs within the pond has a considerable impact, resulting in differences of up to 20% in mass removal. Moreover, the FTI configuration exerts a more pronounced influence on mass removal through FTIs than through sediment deposition alone. In cases where both processes occur simultaneously, the presence of FTIs lead to higher mass removal, primarily attributed to the FTIs themselves, particularly in the initial segment. Remarkably, certain FTI configurations enable mass removal exceeding 70% for large sediment particles, even with a pond length less than half of the original.

Effects of Shear Stress Waves on Meat Tenderness: Ultrasonoporation.

Meat is an important part of the food pyramid in Mexico, to such an extent that it is included in the basic food basket. In recent years, there has been great interest in the application of so-called emerging technologies, such as high-intensity ultrasound (HIU), to modify the characteristics of meat and meat products. The advantages of the HIU in meat such as pH, increased water-holding capacity, and antimicrobial activity are well documented and conclusive. However, in terms of meat tenderization, the results are confusing and contradictory, mainly when they focus on three HIU parameters: acoustic intensity, frequency, and application time. This study explores via a texturometer the effect of HIU-generated acoustic cavitation and ultrasonoporation in beef (m. Longissimus dorsi). Loin-steak was ultrasonicated with the following parameters: time tHIU = 30 min/each side; frequency fHIU = 37 kHz; acoustic intensity IHIU = ~6, 7, 16, 28, and 90 W/cm2. The results showed that acoustic cavitation has a chaotic effect on the loin-steak surface and thickness of the rib-eye due to Bjerknes force, generating shear stress waves, and acoustic radiation transmittance via the internal structure of the meat and the modification of the myofibrils, in addition to the collateral effect in which the collagen and pH generated ultrasonoporation. This means that HIU can be beneficial for the tenderization of meat.

Missing links in preeclampsia cell model systems of endothelial dysfunction.

Preeclampsia, one of the main hypertensive disorders of pregnancy, is associated with circulating factors released by the ischemic placenta accompanied by systemic endothelial dysfunction. The etiology of preeclampsia remains poorly understood although it is associated with high maternal and fetal mortality and increased cardiovascular disease risk. Most cell model systems used for studying endothelial dysfunction have not taken into account hemodynamic physical factors such as shear-stress forces which may prevent extrapolation of cell data to in vivo settings. We overview the role of hemodynamic forces in modulating endothelial cell function and discuss strategies to reproduce this biological characteristic in vitro to improve our understanding of endothelial dysfunction associated with preeclampsia.

Piezo1 activation induces relaxation of the pudendal artery and corpus cavernosum.

Piezo1 channel is a sensor for shear-stress in the vasculature. Piezo1 activation induces vasodilation, and its deficiency contributes to vascular disorders, such as hypertension. In this study, we aimed to determine whether Piezo1 channel has a functional role in the dilation of pudendal arteries and corpus cavernosum (CC). For this, male Wistar rats were used, and the relaxation of the pudendal artery and CC was obtained using the Piezo1 activator, Yoda1, in the presence and absence of Dooku (Yoda1 antagonist), GsMTx4 (non-selective mechanosensory channel inhibitor) and L-NAME (nitric oxide synthase inhibitor). In the CC, Yoda1 was also tested in the presence of indomethacin (non-selective COX inhibitor) and tetraethylammonium (TEA, non-selective potassium channel inhibitor). The expression of Piezo1 was confirmed by Western blotting. Our data show that Piezo1 activation leads to the relaxation of the pudendal artery and CC as the chemical activator of Piezo1, Yoda1, relaxed the pudendal artery (47%) and CC (41%). This response was impaired by L-NAME and abolished by Dooku and GsMTx4 in the pudendal artery only. Indomethacin and TEA did not affect the relaxation induced by Yoda1 in the CC. Limited tools to explore this channel prevent further investigation of its underlying mechanisms of action. In conclusion, our data demonstrate that Piezo1 is expressed and induced the relaxation of the pudendal artery and CC. Further studies are necessary to determine its role in penile erection and if erectile dysfunction is associated with Piezo1 deficiency.

Replicating endothelial shear stress in organ-on-a-chip for predictive hypericin photodynamic efficiency.

Photodynamic therapy using Hypericin (Hy-PDT) is an alternative non-invasive treatment that enables selective tumor inhibition and angiogenesis derived from the differential recruitment of endothelial cells in the tumor microenvironment. Most PDT studies were performed on in vitro models without vascular biomechanical simulation. Our work strives to develop a microchip that generates a constant shear stress force to investigate the Hy-PDT efficiency on human umbilical vein endothelial cells (HUVECs). The microchip with a single straight microchannel was composed of the bottom layer (polystyrene), the middle layer (double-sided biocompatible adhesive tape), and the top layer (polyester film) and could produce shear stress in the range of 1.4 - 7.0 dyn cm-2. The quantification of vascular endothelial growth factor (VEGF), cell viability, and activities of caspases 3 and 7 were assayed to validate the microchip and Hy-PDT efficacy. After the endothelization, static and dynamic cell incubations with Hy were conducted in microchips. Compared to static systems, the shear stress displayed its effect on the increasing release of VEGF and promoted more cell damage and cell death via necrosis during Hy-PDT. In conclusion, the expressive shear stress-dependent manner during PDT treatments suggests that the microchip could be an essential approach in preclinical tests to evaluate the therapeutic outcome considering the endothelial shear stress microenvironment.

Relation of shear stress and KLa on bikaverin production by Fusarium oxysporum CCT7620 in a bioreactor.

This study involved evaluating the effects of rotational impeller speed agitation (N) and specific air flow rate (Фair) on bikaverin production and on the growth of Fusarium oxysporum employing 11 bench-scale bioreactor assays. The results showed that the maximum bikaverin production (close to 300 mg L-1) was achieved after 48 h of fermentation in rice medium (20 g L-1 milled rice in water) at 28 °C with a volumetric oxygen transfer coefficient (KLa) and shear stress values of approximately 20 h-1 and 17 N m-2, respectively. We reached this combination of parameters using an N of 340 rpm and Фair of 0.935 vvm. These KLa and shear stress values can be used as references when upscaling this process. Thus, this study was important to demonstrate how the main parameters in bioreactors affect bikaverin production and it presented important indications for upscaling this bioprocess.

Metabolic effects of CoCr-enriched medium on shear-stressed endothelial cell and osteoblasts: A possible mechanism involving a hypoxic condition on bone healing.

Cobalt-chromium (CoCr)-based alloys have emerged as an interesting biomaterial within biomedical field, mainly considering their biocompatibility, resistance to corrosion and absence of magnetism; however, its effect on cell metabolism is barely known and this prompted us better evaluating whether CoCr-enriched medium affects the metabolism of both osteoblast and endothelial cells, and also if there is a coupling between them. This is also considered here the already-known effect of Cobalt (Co) as a hypoxic element. Firstly, discs of CoCr [subjecting (W) or not (Wo) to dual acid-etched (DAE)] were incubated into FBS-free cell culture medium up to 24 h (37 °C). This CoCr-enriched medium was further used to treat shear-stressed endothelial cells cultures up to 72 h. Thereafter, the conditioned medium containing metabolites of shear-stressed endothelial cells in response to CoCr-enriched medium was further used to subject osteoblast's cultures, when the samples were properly harvested to allow the analysis of the molecular issues. Our data shows that CoCr-enriched medium contains 1.5 ng-2.0 ng/mL of Co, which was captured by endothelial cells and osteoblasts in about 30% in amount and it seems modulate their metabolic pathways: shear-stressed endothelial cells expressed higher profile of HIF1α, VEGF and nNOS genes, while their global profile of protein carbonylation was lower than the control cultures, suggesting lower oxidative stress commitment. Additionally, osteoblasts responding to metabolites of CoCr-challenged endothelial cells show dynamic expression of marker genes in osteogenic differentiation, with alkaline phosphatase (ALP), osteocalcin, and bone sialoprotein (BSP) genes being significantly increased. Additionally, tensional shear-stress forces decrease the stimulus for ColA1gene expression in osteoblasts responding to endothelial cells metabolites, as well as modifying the extracellular matrix remodeling related genes. Analyzing the activities of matrix metalloproteinases (MMPs), the data shows that shear-stressed endothelial cells metabolites increase the activities of both MMP9 and MMP2 in osteoblasts. Altogether, our data shows for the first time that shear-stressed endothelial metabolites responding to CoCr discs contribute to osteogenic phenotype in vitro, and this predicts an active crosstalk between angiogenesis and osteogenesis during osseointegration of CoCr alloy and bone healing, maybe guided by the Co-induced hypoxic condition.

Hazardous cyanobacteria integrity response to velocity gradient and powdered activated carbon in water treatment plants.

Although some studies have investigated the impact caused by chemicals used on water treatment (coagulants and oxidants) on cyanobacteria integrity, the isolated effect of shear stress during coagulation is still not fully understood. This study evaluated the impact of different velocity gradients, mixing times, and the addition of powdered activated carbon (PAC) on the integrity of Microcystis aeruginosa, Raphidiopsis raciborskii, and Dolichospermum circinale, known producers of toxin and taste and odor (T&O) compounds. No association was found between R. raciborskii cell lysis and velocity gradient, with or without PAC, demonstrating the high resilience of this taxon to shear stress. In contrast, an association was found for M. aeruginosa at the highest velocity gradient evaluated (1000 s-1) and for D. circinale above the lowest velocity gradient studied (600 s-1). After PAC addition, there was a reduction in the chances of finding M. aeruginosa intact cells above velocity gradient 800 s-1 at 45 s, while D. circinale show cell lysis in all the scenarios expect at 600 s-1 and 10 s of agitation. The additional impact of PAC on cell lysis may lead to more release of metabolites and shows the need to adjust the hydraulic conditions in the rapid mixing stage, especially when more "fragile" cyanobacteria are present. Neither cyanobacterial cell size nor morphology was shown to be relevant to shear stress sensitivity, indicating that cell wall composition might have been an important factor in controlling cell lysis.

Rapid Fabrication of Microfluidic Devices for Biological Mimicking: A Survey of Materials and Biocompatibility.

Microfluidics is an essential technique used in the development of in vitro models for mimicking complex biological systems. The microchip with microfluidic flows offers the precise control of the microenvironment where the cells can grow and structure inside channels to resemble in vivo conditions allowing a proper cellular response investigation. Hence, this study aimed to develop low-cost, simple microchips to simulate the shear stress effect on the human umbilical vein endothelial cells (HUVEC). Differentially from other biological microfluidic devices described in the literature, we used readily available tools like heat-lamination, toner printer, laser cutter and biocompatible double-sided adhesive tapes to bind different layers of materials together, forming a designed composite with a microchannel. In addition, we screened alternative substrates, including polyester-toner, polyester-vinyl, glass, Permanox® and polystyrene to compose the microchips for optimizing cell adhesion, then enabling these microdevices when coupled to a syringe pump, the cells can withstand the fluid shear stress range from 1 to 4 dyne cm2. The cell viability was monitored by acridine orange/ethidium bromide (AO/EB) staining to detect live and dead cells. As a result, our fabrication processes were cost-effective and straightforward. The materials investigated in the assembling of the microchips exhibited good cell viability and biocompatibility, providing a dynamic microenvironment for cell proliferation. Therefore, we suggest that these microchips could be available everywhere, allowing in vitro assays for daily laboratory experiments and further developing the organ-on-a-chip concept.

Should Heartbeats/Training Session Be Considered When Comparing the Cardiovascular Benefits of High-Intensity Interval Aerobic and Moderate-Intensity Continuous Training? A Critical Appraisal.

The prescription of physical training as a therapeutic measure in the treatment and control of chronic degenerative diseases, mainly cardiovascular disease and metabolic disease, is an increasingly used clinical approach, often preceding the pharmacological prescription. Despite the advances in exercise physiology and cardio functional performance in recent decades, the main challenge is to identify the most appropriate modality, intensity, and training volume for each pathophysiological situation. In this case, the superiority of high-intensity interval training (HIIT) over moderate-intensity continuous training (MICT) has been questioned, since many studies have shown similar results in the different physiological parameters evaluated, especially regarding cardiorespiratory fitness, cardiovascular autonomic control, and cardiac morpho functionality. The cause of conflicting results observed by different studies may be related to standardization, application, and comparison of the two protocols. HIIT would have a higher number of heartbeats compared to MICT, when maintaining high heart rate is disregarded. In this since, our hypothesis for the greatest gains in cardiorespiratory fitness and in the autonomic and cardiovascular adaptations promoted by HIIT is based on the higher volume of training performed as a function of the higher number of heartbeats per unit of time, since the intermittence was calculated based on a percentage of maximum heart rate or reserve heart rate. Nevertheless, the intermittency between the established heart rate percentages is not necessarily accompanied by the intermittent heart rate. Therefore, considering and matching the number of heartbeats performed per training session in both models seems to be a more appropriate way to compare the two training protocols.

LncRNA HOTAIR is a novel endothelial mechanosensitive gene.

To better address whether the long noncoding RNAs (lncRNAs) HOTAIR and HOTTIP are mechanosensitive genes, they were investigated in differentially challenged endothelial cells with respect to a circuit of tensional forces, considering the performance of both arterial and venous endothelial cells. We subjected arterial- and venous-obtained endothelial cells to a circuit of tensional forces within a shear stress model in vitro. Real-time quantitative polymerase chain reaction analysis indicated that microRNA (miRNA)-related processing machinery is significantly required in shear stressed arterial endothelial cell metabolism, which orchestrates miRNA (small noncoding RNA) involvement, and their involvement suggests lncRNA involvement. Of lncRNAs HOTAIR and HOTTIP, only HOTAIR was mechanosensitive considering both arterial and venous endothelial cells, presenting a positive correlation between methylation signature and gene expression. Thereafter, using bioinformatics tools, lncRNA HOTAIR was predicted to modulate miRNA185, miRNA-21, and miRNA23b downregulation. We compared the values of gene expression with a Pearson's correlation test, and expected correlations were observed for miRNA185 (r = 0.8664), miRNA-21 (r = 0.8605), and miRNA23b (0.9128). Taken together, these findings clearly show that lncRNA HOTAIR responds to the shear stress and emerges as a novel mechanosensitive gene in endothelial cells. Altogether, this understanding of mechanosensitive transcriptional and posttranscriptional control involving HOTAIR can also lead to new forms of therapeutic intervention for various diseases, as well as new strategies for tissue engineering and regenerative medicine.

Wortmannin targeting phosphatidylinositol 3-kinase suppresses angiogenic factors in shear-stressed endothelial cells.

Modifications on shear stress-based mechanical forces are associated with pathophysiological susceptibility and their effect on endothelial cells (EC) needs to be better addressed looking for comprehending the cellular and molecular mechanisms. This prompted us to better evaluate the effects of shear stress in human primary venous EC obtained from the umbilical cord, using an in vitro model to mimic the laminar blood flow, reaching an intensity 1-4 Pa. First, our data shows there is a significant up-expression of phosphatidylinositol 3-kinase (PI3K) in shear-stressed cells culminating downstream with an up-phosphorylation of AKT and up-expression of MAPK-ERK, concomitant to a dynamic cytoskeleton rearrangement upon integrin subunits (α4 and ß 3) requirements. Importantly, the results show there is significant involvement of nitric oxide synthase (eNOS), nNOS, and vascular endothelial growth factors receptor 2 (VEGFR2) in shear-stressed EC, while cell cycle-related events seem to being changed. Additionally, although diminution of 5-hydroxymethylcytosine in shear-stressed EC, suggesting a global repression of genes transcription, the promoters of PI3K and eNOS genes were significantly hydroxymethylated corroborating with their respective transcriptional profiles. Finally, to better address, the pivotal role of PI3K in shear-stressed EC we have revisited these biological issues by wortmannin targeting PI3K signaling and the data shows a dependency of PI3K signaling in controlling the expression of VGFR1, VGFR2, VEGF, and eNOS, once these genes were significantly suppressed in the presence of the inhibitor, as well as transcripts from Ki67 and CDK2 genes. Finally, our data still shows a coupling between PI3K and the epigenetic landscape of shear-stressed cells, once wortmannin promotes a significant suppression of ten-11 translocation 1 (TET1), TET2, and TET3 genes, evidencing that PI3K signaling is a necessary upstream pathway to modulate TET-related genes. In this study we determined the major mechanotransduction pathway by which blood flow driven shear stress activates PI3K which plays a pivotal role on guaranteeing endothelial cell phenotype and vascular homeostasis, opening novel perspectives to understand the molecular basis of pathophysiological disorders related with the vascular system.

Patrón de flujo en "4D Flow" y su vínculo con la aortopatía bicúspide / 4D Flow pattern and its role in bicuspid aortic valve aortopathy

ABSTRACT: Bicuspid aortic valve (BAV) disease is generally associated with thoracic aortic dilatation (TAD). Related factors include; genetical, morphological (valvular phenotype) and most recently, hemodynamic profiles associated with flow pattern and wall shear stress. Cardiac magnetic resonance 4D Flow (4DF) can give an integral evaluation of these later flow variables. Remarkable, different spectrums of flow and vortex direction exist in BAV that are related to the site of TAD (proximal or distal). Therefore, we present a 57 years old patient with BAV (Sievers 0) with anteroposterior leaflets distribution in which 4DF depicted an anteriorly and righthand oriented jet that correlated with the zone of grater AD; also, vortex rotation was counterclockwise, corresponding to the most frequent vortex type in BAV. In conclusion, 4DF is a powerful and ground-breaking tool that enhances our knowledge of BAV related aortopathy.

Characterization of the Metabolic Response of Streptomyces clavuligerus to Shear Stress in Stirred Tanks and Single-Use 2D Rocking Motion Bioreactors for Clavulanic Acid Production.

Streptomyces clavuligerus is a gram-positive filamentous bacterium notable for producing clavulanic acid (CA), an inhibitor of ß-lactamase enzymes, which confers resistance to bacteria against several antibiotics. Here we present a comparative analysis of the morphological and metabolic response of S. clavuligerus linked to the CA production under low and high shear stress conditions in a 2D rocking-motion single-use bioreactor (CELL-tainer ®) and stirred tank bioreactor (STR), respectively. The CELL-tainer® guarantees high turbulence and enhanced volumetric mass transfer at low shear stress, which (in contrast to bubble columns) allows the investigation of the impact of shear stress without oxygen limitation. The results indicate that high shear forces do not compromise the viability of S. clavuligerus cells; even higher specific growth rate, biomass, and specific CA production rate were observed in the STR. Under low shear forces in the CELL-tainer® the mycelial diameter increased considerably (average diameter 2.27 in CELL-tainer® vs. 1.44 µm in STR). This suggests that CA production may be affected by a lower surface-to-volume ratio which would lead to lower diffusion and transport of nutrients, oxygen, and product. The present study shows that there is a strong correlation between macromorphology and CA production, which should be an important aspect to consider in industrial production of CA.

Integrated proteomics and metabolomics analysis reveals differential lipid metabolism in human umbilical vein endothelial cells under high and low shear stress.

Atherosclerotic plaque development is closely associated with the hemodynamic forces applied to endothelial cells (ECs). Among these, shear stress (SS) plays a key role in disease development since changes in flow intensity and direction could stimulate an atheroprone or atheroprotective phenotype. ECs under low or oscillatory SS (LSS) show upregulation of inflammatory, adhesion, and cellular permeability molecules. On the contrary, cells under high or laminar SS (HSS) increase their expression of protective and anti-inflammatory factors. The mechanism behind SS regulation of an atheroprotective phenotype is not completely elucidated. Here we used proteomics and metabolomics to better understand the changes in endothelial cells (human umbilical vein endothelial cells) under in vitro LSS and HSS that promote an atheroprone or atheroprotective profile and how these modifications can be connected to atherosclerosis development. Our data showed that lipid metabolism, in special cholesterol metabolism, was downregulated in cells under LSS. The low-density lipoprotein receptor (LDLR) showed significant alterations both at the quantitative expression level as well as regarding posttranslational modifications. Under LSS, LDLR was seen at lower concentrations and with a different glycosylation profile. Finally, modulating LDLR with atorvastatin led to the recapitulation of a HSS metabolic phenotype in EC under LSS. Altogether, our data suggest that there is significant modulation of lipid metabolism in endothelial cells under different SS intensities and that this could contribute to the atheroprone phenotype of LSS. Statin treatment was able to partially recover the protective profile of these cells.

Cell-surface HSP70 associates with thrombomodulin in endothelial cells.

Heat shock protein-70 (HSP70) is crucial for proteostasis and displays cell-protective effects. Meanwhile, enhanced levels of cell-surface (cs) and secreted HSP70 paradoxically associate with pathologic cardiovascular conditions. However, mechanisms regulating csHSP70 pool are unknown. We hypothesized that total and csHSP70 expressions are modulated by hemodynamic forces, major contributors to endothelial pathophysiology. We also investigated whether thrombomodulin, a crucial thromboresistance cell-surface protein, is a csHSP70 target. We used proteomic/western analysis, confocal microscopy, and cs-biotinylation to analyze the pattern and specific characteristics of intracellular and csHSP70. HSP70 interaction with thrombomodulin was investigated by confocal colocalization, en face immunofluorescence, proximity assay, and immunoprecipitation. Thrombomodulin activity was assessed by measured protein C activation two-step assay. Our results show that csHSP70 pool in endothelial cells (EC) exhibits a peculiar cluster-like pattern and undergoes enhanced expression by physiological arterial-level laminar shear stress. Conversely, total and csHSP70 expressions were diminished under low shear stress, a known proatherogenic hemodynamic pattern. Furthermore, total HSP70 levels were decreased in aortic arch (associated with proatherogenic turbulent flow) compared with thoracic aorta (associated with atheroprotective laminar flow). Importantly, csHSP70 co-localized with thrombomodulin in cultured EC and aorta endothelium; proximity ligation assays and immunoprecipitation confirmed their physical interaction in EC. Remarkably, immunoneutralization of csHSP70 enhanced thrombomodulin activity in EC and aorta ex vivo. Overall, proatherogenic hemodynamic forces promote reduced total HSP70 expression, which might implicate in disturbed proteostasis; meanwhile, the associated decrease in cs-HSP70 pool associates with thromboresistance signaling. Cell-surface HSP70 (csHSP70) expression regulation and csHSP70 targets in vascular cells are unknown. We showed that HSP70 levels are shear stress-modulated and decreased under proatherogenic conditions. Remarkably, csHSP70 binds thrombomodulin and inhibits its activity in endothelial cells. This mechanism can potentially explain some deleterious effects previously associated with high extracellular HSP70 levels, as csHSP70 potentially could restrict thromboresistance and support thrombosis/inflammation in stress situations.

c-Src kinase contributes on endothelial cells mechanotransduction in a heat shock protein 70-dependent turnover manner.

Shear stress changes are associated with a repertory of signaling cascade modulating vascular phenotype. As shear stress-related tensional forces might be associated with pathophysiological susceptibility, a more comprehensive molecular map needs to be addressed. Thus, we subjected human umbilical vein endothelial cells (HUVECs) to a circuit of different tensional forces in vitro considering the following three groups: (a) physiological blood flow shear stress condition (named Normo), (b) a hypertensive blood flow shear stress (named Hyper), and (c) these hyper-stressed cells were returned to Normo condition (named Return). The samples were properly collected to allow different methodologies analysis. Our data showed a pivotal involvement of c-Src on driving the mechanotransduction cascade by modulating signaling related with adhesion, survival (PI3K/Akt) and proliferative phenotype. Moreover, c-Src seems to develop important role during extracellular matrix remodeling. Additionally, proteomic analysis showed strong involvement of heat shock protein 70 (HSP70) in the hypertensive-stressed cells; it being significantly decreased in return phenotype. This result prompted us to investigate 20S proteasome as an intracellular proteolytic alternative route to promote the turnover of those proteins. Surprisingly, our data reveled significant overexpression of sets of proteasome subunit α-type (PSMA) and ß-type (PSMB) genes. In conjunction, our data showed c-Src as a pivotal protein to drive mechanotransduction in endothelial cells in a HSP70-dependent turnover scenario. Because shear patterns is associated with pathophysiological changes, such as atherosclerosis and hypertension, these results paved new road to understand the molecular mechanism on driving mechanotransduction in endothelial cells and, if drugable, these targets must be considered within pharmacological treatment optimization.

Modelo matemático simplificado para determinar a capacidade erosiva de um escoamento lamoso em regime laminar pulsante / A simplified mathematical model to estimate the erosive capacity of a pulsating laminar muddy flow

RESUMO Neste artigo foi proposto um modelo matemático simplificado para inferir sobre a capacidade erosiva de um escoamento lamoso em regime laminar com presença de roll waves (efeito pulsante) na superfície livre. Na perspectiva de aplicação de resultados no contexto de desastres naturais (corridas de lama), foi determinada a evolução da velocidade média e da tensão de cisalhamento no fundo do canal. O modelo proposto é baseado nas equações de águas rasas e na reologia de Herschel-Bulkley, como representativa das lamas. A validação e a aplicação do modelo foram feitas a partir de dados experimentais de roll waves geradas em canal no laboratório confrontados com resultados de simulações numéricas. Os resultados obtidos mostram um incremento na capacidade erosiva, na presença de roll waves, da ordem de 12% em relação ao escoamento base, valor este em concordância com os 10% apontados por Ng e Mei (1994) para uma reologia mais simplificada.

ABSTRACT In this paper, we proposed and validated a simplified mathematical model to estimate the erosive capacity of a muddy laminar flow with roll waves (pulsating effect) on the free surface. From the perspective of applying results in the context of natural disasters (mudflows), the evolution of mean velocity and bottom shear stress in the channel were determined. The proposed model is based on shallow water equations and Herschel-Bulkley rheology as representative of the muds. The model was validated by comparison with results of numerical simulations and experimental data of roll waves generated in a channel. In-depth comparative analyses were made for inferring the erosive capacity of this sort of flow. The obtained results show an increase of erosive capacity (12%) due to the presence of roll waves in relation to the uniform flow's. Similar values (10%) were found by Ng and Mei (1994) for a more simplified rheology model.