bluuebunny/crossref_metadata_2025_split huggingface dataset:
Aneurysmal cyclic circumferential stretch affects the expression of endothelial genes involved in vascular homeostasis
Authors: Diagbouga MR , Morel S , Lemeille S , Kwak BR 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.

Background
Intracranial aneurysm (IA) rupture is a common cause of hemorrhagic stroke. IAs are present in 3 to 5% of the general population and the annual rupture rate of an IA has been estimated to 1%. Once formed, an IA can rupture immediately, grow before rupturing, or remain stable. Predicting the evolution of an unruptured IA is very difficult. Indeed, the biology of the IA wall is complex and the precise biomechanical processes leading to aneurysm wall rupture are not yet known. While the contribution of wall shear stress in IA disease has been demonstrated in several studies, the role of cyclic circumferential stretch (CCS) remains poorly understood.

Purpose
The levels of CCS in saccular IAs are unknown but are generally considered very low or even absent. The goal of our study was to investigate the transcriptional response of endothelial cells (ECs) to absence of CCS and the potential impact on the endothelium function.

Methods
Human umbilical veins ECs (HUVECs) from 6 different donors were exposed to physiological CCS levels for cerebral arteries (6% stretch) or to aneurysmal CCS (0%) for 48h using the Flexcell strain unit FX-5000T, and unbiased transcriptomics were performed. The expression of selected genes was confirmed by qPCR.

Results
As a validation of our methodology, we verified the induction of eNOS phosphorylation after exposure of the ECs to physiological CCS. Stretched EC samples display a high similarity between each other while EC samples under aneurysmal CCS were spread-out on a multidimensional scaling plot. The differential gene expression of ECs exposed to aneurysmal or physiological CCS revealed 51 up-regulated genes and 49 down-regulated genes in absence of CCS. The gene set enrichment analysis identified 70 up-regulated and 38 down-regulated pathways in HUVECs exposed to aneurysmal CCS. Up-regulated pathways were involved in oxidative stress, angiogenic and inflammatory pathways, and the down-regulated pathways in proliferation and extracellular matrix-receptor interactions. We confirmed by qPCR the down-regulation by aneurysmal CCS of Gja4 and Gja5, genes coding for the gap junction proteins Connexin37 (Cx37) and Connexin40 (Cx40), respectively. Connexins (Cxs) are crucial proteins for endothelial homeostasis, vascular function, endothelial cell cycle regulation and inflammation, and their role in the context of IA disease will be further studied.

Conclusion
Our results indicate that physiological CCS prevents large deviation in ECs and regulate the expression of genes essential for maintaining vascular homeostasis. The down-regulation of endothelial Cx37 and Cx40 under aneurysmal CSS may contribute to the endothelial dysfunction associated with IA progression. Further investigation may help to better define the role of Cxs in aneurysmal disease and to understand the significance of absence of CCS in IA disease progression.
Mitochondrial division inhibitor 1 (mdivi-1) as a potential pharmacological strategy to prevent vascular restenosis
Authors: Crespo-Avilan G , Hernandez-Resendiz S , Ramachandra C , Liehn E , Preissner K , Hausenloy D 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Singapore National Medical Research Council

Background
New treatments are needed to prevent post-angioplasty and stent restenosis in coronary artery disease (CAD) and peripheral arterial disease (PAD) patients. Accumulated findings have shown that mitochondrial dysfunction is involved in the pathophysiology mechanisms underlying several cardiovascular conditions, however, in the scenario of vascular remodelling after injury, the role of mitochondrial dynamics is not completely understood.

Purpose
In this study, we investigate the effect of mdivi-1 on neointimal hyperplasia and plaque development in a wire-induced endothelial injury animal model.­

Methods
Using ApoE-/- mice on a background of hyperlipidaemia, we investigated weather in vivo administration of mdivi-1 (1.2mg/kg/d) could reduce atherosclerotic plaque volume, plaque complexity and inflammation in a carotid-wire injury model of neointimal hyperplasia. Also, THP-1 monocytes (T-M) and THP-1-derived Macrophages (T-DM) were stimulated in vitro with LPS + IFN-gamma to induce M1 polarization and then subjected to gene expression and protein analysis in the presence or absence of mdivi-1 (50µM). Transmigration and 3D chemotaxis assays were performed in T-M cells and Human peripheral blood monocytes, to evaluate the effect of mdivi-1 on cell migration in response to M-CSF and MCP-1. Finally, we investigated the effects of Mdivi-1 on mitochondrial respiration using Seahorse assay in T-M and T-DM.

Results
In vivo treatment with mdivi-1 reduced neointimal hyperplasia by 37% when compared to control, and was associated with a significant decrease of vascular smooth muscle cells and macrophages cell numbers, as well as reduction of the pro-inflammatory mediators TNF-α and ICAM-1 in the plaque. In vitro M1 polarization of T-M and T-DM induced both up-regulation and production of pro-inflammatory mediators and mdivi-1 significantly suppressed this effects. In addition, monocyte chemotaxis response to M-CSF and MCP-1 was significantly reduced in the presence of mdivi-1. Lastly, mdivi-1 reduced oxygen consumption rate in T-M and T-DM, and prevented M1-cell polarization.

Conclusions
We show that mdivi-1 significantly reduces vessel wall thickness, neointimal hyperplasia, monocyte recruitment, and inflammation after vascular endothelial injury, positioning Mdivi-1 as a potential pharmacological strategy to reduce restenosis following angioplasty and stenting in CAD and PAD patients.
Comparative secretome analysis of obese perivascular adipose tissue reveals impaired adipose-neuronal crosstalk
Authors: Duregotti E , Reumiller C , Mayr U , Hasman M , Schmidt L , Burnap S A , Theofilatos K , Barallobre-Barreiro J , Viviano A , Jahangiri M , Mayr M 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Foundation. Main funding source(s): BHF Programme Grant (RG/16/14/32397)
VASCage-Research Center on Vascular Ageing and Stroke (No. 868624)

Background
While canonical adipose tissue (AT) depots have been extensively characterised in health and disease, comparatively little is known about the pathological changes affecting the perivascular AT’s (PVAT) physiology during obesity.

Purpose
The aim of this study was to study the impact of obesity on the secretory activity of the murine PVAT.

Methods
We exploited proteomics to profile the secretome of peri-aortic and canonical AT depots in wild-type (wt) and obese (ob/ob) mice. In parallel, fat tissues were processed for biochemical and histological analysis and mechanistical experiments were performed in vitro on primary neuronal cultures.

Results
Proteomics on ATs conditioned media from wt mice revealed that each fat depot displays a unique secretory profile. The enrichment of neuronal cell-adhesion molecules detected in PVAT secretomes reflected a higher content of intra-parenchymal sympathetic projections compared to non-perivascular ATs. A significant decrease of the same neuronal proteins in PVAT conditioned media from ob/ob mice was found to be associated with a substantial reduction of the perivascular sympathetic innervation. Intriguingly, a similar decrease of sympathetic markers was detected in the epicardial AT from obese patients. Mechanistically, the conditioned media from ob/ob AT explants was found to exert a deleterious effect on the axons of primary sympathetic neurons in vitro, indicating that this neuropathy is due to local alterations of the PVAT secretome that detrimentally impact on the embedded sympathetic neurites. Among proteins significantly down-regulated in the secretomes of ob/ob PVAT samples, neuronal growth regulator 1 (Negr1) was found to promote axonal elongation and branching on sympathetic neurons in vitro. Administration of recombinant Negr1 also partially restored the neurotrophic effect of ob/ob AT secretomes on sympathetic axons both in vitro and in vivo.

Conclusions
Obesity-related alterations in the secretome of PVAT severely affect the homeostasis of the perivascular environment, leading to a loss of perivascular sympathetic innervation. A novel neurotrophic role is unveiled for Negr1, whose locus has been associated with human obesity.
MicroRNAs in circulating microvesicles and plasma as biomarkers that complement the clinical diagnosis of diabetic dyslipidemia and its complications
Authors: Nemecz M , Stefan DS , Constantin A , Procopciuc A , Comarita K , Tanko G , Georgescu A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Research funded by grants of the Romanian Ministry of Education and Research, CNCS-UEFISCDI, project no. PN-III-P1-1.2-PCCDI-2017–0527 (Contract no. 83PCCDI/2018), project no. PN-III-P1-1.1-TE-2019–0811, within PNCDI III” (Contract no. TE 97/2020)

Background
Dislypidemia is a major risk factor for cardiovascular diseases as well as for diabetes where often leads to severe microvascular complications. Reportedly, microRNAs (miRNAs) associated with microvesicles (MVs) regulate cellular mechanisms in the progression of diabetic dyslipidemia. We hypothesised that detection of the changes in plasma MV components could be used as early biomarkers to prevent the irreversible effects of diabetic dyslipidemia. Purpose: of this study was to search whether in diabetic dyslipidemia with/without microvascular complications there are modifications of the circulating MVs and plasma miRNAs and whether these changes could be predictive of the disease evolution.

Methods
Plasma/sera was collected from diabetic patients with dyslipidemia with/without microvascular complications and the main parameters and the inflammatory markers were assessed. Results showed that, compared to either diabetes or dyslipidemia alone, in the diabetic dyslipidemic patients, significant increases in plasma glucose, cholesterol, LDL-cholesterol, triglyceride concentrations, systolic blood pressure were present. In addition, high levels of circulating IL-6 and MVs-derived from endothelial cells (CD144+), platelets (CD41+), macrophages (CD14+), and leucocytes (CD18+) were detected. Significant changes were found in plasma miRNAs and MV-associated miRNAs: 122,-132,-143,-200b,-212, decreased miR-34a,-155,-218 expressions in MVs, and reduced miR-21,-34a,-132,-143 expressions in plasma. Notably, when microvascular complications were associated to diabetic dyslipidemia, we found significant changes such as increased expression of miR-21,-34a,-122,-126,-143,-218,-223 in MV and plasma miR-143,-132 and decreased miR-200b,-212 in MVs and plasma miR-223.

Conclusion
In patients with diabetic dyslipidemia and, in particular, with diabetic dyslipidemia associated with microvascular complications, a significant modification in the plasma MV content and miRNAs occur suggesting that they could turn into biomarkers and potential diagnostic tool for diabetic dyslipidemia and its complications.
Alterations in coronary resistance artery network geometry in diabetes and the role of Tenascin C
Authors: Aykac I 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.
Background and Aims: Geometrical alterations in the coronary resistance artery network and potential involvement of the Tenascin C (TNC) protein were examined in diabetic mice.

Methods
Diabetes was induced by streptozotocin injections (n=7-11 animals in each group) in Wild type (A/J) and Tenascin C KO (TNC KO) mice, and 16-18 weeks later the heart was removed and the whole subsurface network of the left coronary artery was microprepared (down to branches of 40 µm outer diameter), in situ pressure-perfused and studied using video-microscopy. Outer and inner diameters, wall thicknesses and bifurcation angles were measured on whole network pictures reconstructed into collages at 1.7 µm pixel resolutions.

Results
Diabetic networks featured abnormal morphological alterations including trifurcations, sharp bends of larger branches, and branches directed in the retrograde direction (p&lt;0.001 by the χ2 test). Networks of TNC KO mice tended to form early divisions producing parallel running larger branches (p&lt;0.001 by the χ2 probe). Diabetic networks were substantially more abundant in 100-180 µm components, appearing in 2-5 mm flow distance from orifice. This was accompanied by thickening of the wall of larger arterioles (&gt;220 µm) and thinning of the wall of a population of smaller (100-140 µm) arterioles (p&lt;0.001). Interestingly, diabetes model by STZ-injection did not induce further geometrical changes in TNC KO mice. Blood flow should cover larger distances in diabetic networks.

Conclusion
In diabetic mice, a combined network remodelling of the coronary vasculature was observed with hypertrophic and hypotrophic remodelling and vasculogenesis at well defined, specific positions of the network. TNC plays an important role in the formation of network geometry, and TNC knockout induces parallel fragmentation preventing diabetes-induced abnormal vascular morphologies
Transition for transduction: how to make cardiac endothelial cells more permissive to AAV
Authors: Volf N , Ring N , Colliva A , Vuerich R , Braga L , Zentilin L , Zacchigna S 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.

Introduction
Despite major attempts aiming to revascularize an ischemic heart, prognosis of cardiac ischemia remains poor. A major obstacle for the induction of new blood vessels, referred as therapeutic angiogenesis, is inefficient gene transfer in cardiac endothelial cells (ECs). This poor permissiveness to genetic modification is a limiting step in studies of pathological conditions with inadequate, but also with increased vascularization. To study the function of a specific gene and its potential implications in angiogenesis, the most straightforward approach would be to either overexpress or silence it in ECs. We and others have tested multiple protocols for either transfection and transduction of primary ECs in vitro and in vivo, with no success. Vectors based on Adeno-associated virus (AAV) and Lentivirus (LV) stand as ideal tools for gene delivery, however they do not work in cardiac ECs.

Purpose
To enhance the permissiveness of EC, we optimized and performed high-throughput screening on adult cardiac ECs using a library of FDA approved drugs, to search for compounds increasing transduction by AAV6 and LV, with the final aim to identify the major determinants inhibiting genetic modification of ECs.

Methods
Cardiac ECs were isolated from 2 months old C57BL/6 mice and plated on 384-well plates. After 48 hours, the FDA drug library (&gt; 1500 compounds) was added together with AAV6-dsRed and LV-GFP. Cells were fixed with 4% PFA 72 hours post-transduction, and subsequently stained for the EC marker ERG. The levels of AAV6-dsRed, LV-GFP and ERG expression were assessed by automated, high-content microscopy.

Results
After two rounds of screening at different drug concentrations, we obtained a list of candidate compounds able to increase adult cardiac EC transduction by either AAV6 or LV over two folds. Among these, our best hit increasing AAV6-mediated transduction was Vatalanib, a tyrosine kinase inhibitor acting on VEGFRs, PDGFR, and c-Kit. Valatanib not only increased permissiveness to AAV6, but it also changed the morphology of the cells, which became elongated and similar to mesenchymal cells. This suggested ongoing endothelial mesenchymal transition (EndMT), which was verified by decreased CD31 and increased α-SMA expression. Interestingly, AAV mostly transduced cells with the lowest CD31 expression.

Conclusions
Our high-throughput screening identified Vatalanib as the most potent drug in increasing permissiveness of cardiac ECs to AAV transduction. Because the drug induced loss of CD31 and gain in α-SMA, we hypothesized that the reason for increased permissiveness was EndMT. This hypothesis is consistent with the known mechanism of action of Valatanib, which inhibits VEGFR2 and thus interferes with VEGF signalling, that in turn is essential to maintain EC identity.
Correction of myocardial changes on an animal experimental model with meldonim
Authors: Yarmolenko O , Bumeister V , Prykhodko O , Gordienko O , Demikhova N 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: None.
Prolonged hypoosmolar overhydration causes a volume overload necessitating cardioprotectors together with a complex of measures aimed at stabilizing the water and electrolyte balance. The content of water and electrolytes in the human body changes throughout life, so the body's response to the disorder of water and salt balance may be different depending on age. The fluid overload positively correlates with adverse outcomes in critically ill patients. One of the main tasks of modern cardiology is to find the means of leveling the influence of adverse factors on myocardium in different aged patients.
The aim of our study was the determination of morphological changes in the myocardium of old laboratory animals under the influence of severe hypoosmolar hyperhydration and attempt to correct the identified changes by means of meldonium.

Materials and methods
The experiment was conducted in accordance with the "European Convention of the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes" (Strasbourg, 1986). 18 rats at the age of 22 months were divided into 3 series: 2 experimental and 1 control (6 in each). Animals of the first experimental series were modeling severe hypoosmolar hyperhydration by introduction of 10 ml distilled water three times a day through a tube during 25 days. Animals ate boiled unsalted food and were injected a synthetic analogue of antidiuretic hormone "Minirin" (Ferring) with drinking water 2 times a day at a dose 0.01 mg. In addition, the rats of the second experimental series were injected with meldonium intraperitoneally at a dose of 50 mg/kg once a day throughout the entire experiment. The animals of control series received normal drinking water and food and were injected the "Mynyrin"(Ferring) twice a day at a dose of 0.01 mg, considering the potential effects of vasopressin on the cardiovascular system. We measured the diameter of cardiomyocytes (DCMC) of both ventricles, the relative amount of cardiomyocytes (RACMC), connective tissue (RACT), and vessels (RAV), stromal-parenchymal ratio in the ventricles by the formula (RACT+RAV)/RACMC.

Results
In myocardium of rats administered meldonium, only isolated stromal hemorrhage are noted, while in myocardial samples of the first experimental series there are numerous hemorrhages. DCMC of left ventricle and DCMC of right ventricle are greater than control at 13.18% (p&lt;0.0001) and 12.27% (p&lt;0.0001) respectively. RAV does not change. RACT of left ventricle and RACT of right ventricle increase respectively by 3.72% (p=0.0060) and 5.23% (p &lt;0.0001). RACMC of right ventricle is smaller than the control parameter by 0.63% (p = 0.0015).

Conclusion
In old animals, meldonium improves the vascular component of the myocardium. However, we have not found the positive effect of the drug on other studied parameters.
Vascular graft storage solution preserves endothelial function
Authors: Winkler B , Szabo PL , Dostal C , Arnold Z , Geisler D , Crailsheim I , Folkmann S , Grabenwoeger M , Podesser BK , Kiss A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Karl Landsteiner Institute Cardiovascular Research

Background
Saline is still the most widely used storage and flushing solution in cardiovascular procedures despite knowing evidence of its influence on the human endothelial cell function. Aim of this study was to assess the effect of DuraGraft©, an intraoperative graft treatment solution, on human saphenous vein segments, rat aortic segments and human umbilical vein endothelial cells (HUVECs) in comparison to saline.

Methods
Within 12 patients undergoing aortocoronary bypass surgery, saphenous vein graft segments were randomized to DuraGraft© (n=12/6) or saline (n=12/6) solution before intraoperative storage. These segments as well as rat aortic segments underwent assessment of vascular function in a multichamber isometric myograph system in comparison to Krebs-Henseleit solution (KHS), a physiologic organ buffer solution. Additionally, human umbilical vein endothelial cells (HUVECs) were used for cell viability tests.

Results
KCl-induced contraction showed a tendency toward increase when treated with DuraGraft© compared to normal saline preservation of human vein segments (24.73±16.22 vs. 15.59±9.53 N/m2, P

Conclusion
DuraGraft© demonstrated a favorable effect on graft relaxation and contraction indicating preservation of vascular endothelial function. Saline is clearly not only inferior to this specialized solution but may show additional harmful effects to viability.
Tip detection-antegrade dissection and reentry using intravascular ultrasound in chronic total occlusion intervention: first human case report
Authors: Suzuki Satoshi , Okamura Atsunori , Nagai Hiroyuki , Iwakura Katsuomi 
Published: June 2022

Abstract

Background
It has been considered impossible to perform antegrade dissection and reentry (ADR) by simply puncturing with a guidewire using the intravascular ultrasound (IVUS) observation without the support of the Stingray system.

Case summary
A 78-year-old man suffered from effort angina pectoris due to a chronic total occlusion (CTO) lesion in the left circumflex coronary artery. A retry percutaneous coronary intervention for the CTO lesion was attempted at our hospital. The J-CTO score was 3. The first guidewire entered the subintimal space. We moved onto Stingray-ADR using the angiographic observation, but the guidewire could not be led into the true lumen. AnteOwl WR-IVUS (AO-IVUS) observation revealed a distal true lumen in which the inner lumen was maintained 5 mm beyond the CTO exit. We decided to perform the tip detection-ADR using the AO-IVUS observation. The tip detection method allowed the tip of the penetration wire to puncture the wall between the subintima and the true lumen in an exactly vertical direction, resulting in the successful creation of a reentry point. The CTO lesion was dilated with one drug-eluting stent, and normal antegrade blood flow was achieved.

Discussion
Since the tip detection method enables accurate punctures, it may allow for ADR by simply puncturing using a guidewire. Due to this method being performed using the IVUS observation, it is likely more reliable than Stingray-ADR using the angiographic observation.