Autophagy-related genes analysis reveals potential biomarkers for prediction of the impaired walking capacity of peripheral arterial disease.

BACKGROUND: The role of autophagy and autophagy-related genes in peripheral arterial disease (PAD) remains unknown and may be of diagnostic and prognostic value. The aim of this study is to investigate the relationship between autophagy and PAD, and identify potential diagnostic or prognostic biomarkers for medical practice. METHODS: Differentially expressed autophagy-related genes in PAD were explored from GSE57691 and validated in our WalkByLab registry participants by quantitative real-time polymerase chain reaction (qRT-PCR). The level of autophagy in peripheral blood mononuclear cells (PBMCs) of WalkByLab participants was assessed by analyzing autophagic marker proteins (beclin-1, P62, LC3B). Single sample gene set enrichment analysis (ssGSEA) was used to evaluate the immune microenvironment within the artery wall of PAD patients and healthy persons. Chemokine antibody array and enzyme-linked immunosorbent assay were used to assess the chemokines in participants' plasma. Treadmill testing with Gardner protocol was used to evaluate participants' walking capacity. Pain-free walking distance, maximum walking distance, and walking time were recorded. Finally, a nomogram model based on logistic regression was built to predict impaired walking performance. RESULTS: A total of 20 relevant autophagy-related genes were identified, and these genes were confirmed to be expressed at low levels in our PAD participants. Western blotting demonstrated that the expression of autophagic marker proteins beclin-1 and LC3BII were significantly reduced in PAD patients' PBMCs. ssGSEA revealed that most of the autophagy-related genes were strongly correlated with immune function, with the largest number of associated genes showing interaction between cytokine-and-cytokine receptors (CCR). In this context, the chemokines growth-related oncogene (GRO) and neutrophil activating protein2 (NAP2) are highly expressed in the plasma of WalkByLab PAD patients and were significantly negatively correlated with the walking distance assessed by Gardner treadmill testing. Finally, the plasma NAP2 level (AUC: 0.743) and derived nomogram model (AUC: 0.860) has a strong predictive potential to identify a poor walking capacity. CONCLUSIONS: Overall, these data highlight both the important role of autophagy and autophagy-related genes in PAD and link them to vascular inflammation (expression of chemokines). In particular, chemokine NAP2 emerged as a novel biomarker that can be used to predict the impaired walking capacity in PAD patients.

Usability of Fantom Encore® scaffold in non-complex bifurcations-Analysis in bench models.

OBJECTIVE: Present bench study aimed to evaluate whether technical characteristics of Fantom Encore® bioresorbable scaffold (BRS) allow to perform proximal optimization/side branch dilation/proximal optimization (POT-SB-POT) technique, as an adequate solution for bifurcation percutaneous coronary intervention. METHODS: Two Fantom Encore® BRS platforms (small with 3.0 mm nominal diameter, n = 7; and large with 3.5 mm nominal diameter, n = 7) were evaluated in bench models, which were designed according to Finet-law and fitted to nominal scaffold diameter in the distal main branch (MB) and fitted to indicated maximal expansion capacity in the proximal main branch (MB). Results were evaluated by (a) fluoroscopy, (b) optical coherence tomography (OCT) and (c) micro-computed tomography (µCT). RESULTS: All procedures were performed according to the protocol. Careful review of the fluoroscopic loops by an independent operator did not reveal any strut fracture or major deformation. By OCT the overall rate of perfectly apposed struts in the bifurcation area was 15 ± 6% after SB opening, that increased significantly but remained low with 22 ± 9% after final POT (p = .001). Compared to SB ballooning alone, significant benefit of final POT was found in rate of perfect apposition in the proximal MB (15 ± 12% vs. 26 ± 15%, respectively; p = .017) and at the abostial side of polygon of confluence (7 ± 9% vs. 16 ± 13%, respectively; p = .005). µCT analysis revealed a single strut fracture in one case with the small platform, while four cases showed single or multiple strut fractures with the large platform. CONCLUSION: The mechanical characteristics of the device are not suitable for use of Conventional techniques for bifurcation PCI such as POT-SB-POT. The use of Fantom Encore® BRS for bifurcation PCI with relevant SB should not be encouraged.

Living life with cerebral palsy? A description of the social safety nets for individuals with cerebral palsy in the Nordic countries.

AIMS: This report reviews major laws, acts and regulations of social benefits and services for individuals with disabilities, focusing on cerebral palsy in the five Nordic countries. It summarizes the available benefits and services and the re-application process and provides comparative analyses among the countries. METHODS: Published reports, articles and relevant government and municipal websites were reviewed for each respective country and used to compile an overview and comparison between the countries. RESULTS: In the Nordic countries, there are a number of laws and regulations in place to support individuals with cerebral palsy and their families. In addition, there are numerous social benefits available for which individuals with disabilities can apply. Although there are national differences, the similarities across the five countries regarding laws, social benefits offered for individuals with cerebral palsy and the application processes are clear. However, the application processes seem cumbersome and, at times, redundant. Physicians and other healthcare specialists repeatedly need to write 'medical certificates' describing the diagnosis and its consequences for a disability that is chronic and lifelong. CONCLUSIONS: Participation in society for individuals with cerebral palsy disabilities can be enabled by social benefits. By extension, social benefits may indirectly have implications for public health in individuals with disabilities. Although the lives of individuals with cerebral palsy - as with others - can improve in certain areas, the need for social benefits will generally increase, not decrease, over time. Although it is clearly important to have checks and balances that prevent system misuse, it might be worthwhile from a cost-benefit perspective to investigate whether the current systems could be improved to better manage time and resources and avoid emotional distress by streamlining the application process.

Effect of ACEI and ARB treatment on nitric oxide-dependent endothelial function.

Background: Angiotensin-converting-enzyme inhibitors (ACEI) and angiotensin II receptor blockers (ARB) are widely used as a first-line therapy for the treatment of cardiovascular disease. Here, ACEI modulate the bradykinin receptor (BDKRB1 and BDKRB2) system and NO-dependent endothelial function, thus determining cardiovascular health and regenerative arteriogenesis. The current study aims at evaluating nitric oxide-dependent endothelial function, and gene expression of bradykinin receptors in peripheral blood mononuclear cells (PBMC) from patients with ACEI or ARB treatment. Patients and methods: The WalkByLab has been established to screen cardiovascular patients for peripheral artery disease and coronary artery disease. In total 177 patients from WalkByLab with heterogenous disease and risk status were randomly selected, divided according to their medication history into the following groups: 1. ACEI group, 2. ARB group or 3. non-ACE/ARB group. Total plasma nitrite/nitrate (NO) levels were measured, endothelial function was evaluated by assessing flow meditated dilation (FMD). PBMC were isolated from peripheral whole blood, and gene expression (qRT-PCR) of bradykinin receptors and angiotensin converting enzyme were assessed. Results: Plasma total NO concentration in the ACEI group (24.66±16.28, µmol/l) was increased as compared to the ARB group (18.57±11.58, µmol/l, P=0.0046) and non-ACE/ARB group (16.83±8.64, µmol/l, P=0.0127) in patients between 40 to 90 years of age. However, FMD values (%) in the ACEI group (7.07±2.40, %) were similar as compared to the ARB (6.35±2.13, %) and non-ACE/ARB group (6.51±2.15, %), but significantly negatively correlated with age. Interestingly, BDKRB1 mRNA level was significantly higher and BDKRB2 mRNA level lower in the ACEI group (BDKRB1 3.88-fold±1.05, BDKRB2 0.22-fold±0.04) as compared to the non-ACE/ARB group (BDKRB1 1.00-fold±0.39, P<0.0001, BDKRB2 1.00-fold±0.45, P=0.0136). Conclusions: ACEI treatment enhances total nitrite/nitrate concentration, furthermore, upregulates BDKRB1 in PBMC, but downregulates BDKRB2 mRNA expression. FMD is a strong determinant of vascular aging and is sensitive to underlying heterogenous cardiovascular diseases.

Long-term individual shear rate therapy counterpulsation enhances plasma nitrite release in patients with PAD.

BACKGROUND: Individual shear rate therapy (ISRT) has been designed as a novel non-invasive treatment option for peripheral artery disease (PAD) patients and has been shown to improve endothelial function and walking distance. The aim of this study was to elucidate the impact of ISRT on the level of nitric oxide in patient blood plasma and the expression of related molecular markers in peripheral blood mononuclear cells (PBMCs). Molecular diagnostic tests were performed for two ISRT trials. PATIENTS AND METHODS: In ISRT-1 26 healthy subjects underwent one session of treadmill training and one session of ISRT respectively in a cross-over design. In ISRT-2 14 PAD patients with a stable intermittent claudication underwent a 30 hours long-term treatment. Plasma nitrite release as well as the mRNA expression of NOS2 and key regulators of the kallikrein-kinin system were measured in PBMCs at different time points. RESULTS: Short-term ISRT revealed significantly decreased NOS2 expression in PBMCs of healthy volunteers and PAD patients. Long-term ISRT, in turn, demonstrated a significant plasma nitrite increase in PAD patients. CONCLUSIONS: We verified that long-term ISRT stimulates the vascular system and exerts a comparable effect to physical exercise in regards to NO release, which coincide with recent findings regarding an improvement of endothelial function. However, further studies are necessary to investigate the role for circulating leukocytes.
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Adaptation of external counterpulsation based on individual shear rate therapy improves endothelial function and claudication distance in peripheral artery disease.

BACKGROUND: External counterpulsation therapy enhances blood flow and was shown to improve endothelial function and quality of life in coronary artery disease patients. However, high pressures of up to 300 mmHg may lead to malperfusion of the ischaemic limb. To improve the clinical outcome of patients with peripheral artery disease (PAD), we adjusted external counterpulsation and developed a novel non-invasive approach termed individual shear rate therapy (ISRT). PATIENTS AND METHODS: In the present study, 14 patients with a Fontaine stage IIb and femoral-popliteal PAD underwent 30 hours of ISRT over 5 weeks. For ISRT, individual treatment pressures that do not exceed 160 mmHg were assessed by Doppler flow parameters during counterpulsation (individual shear rate diagnosis) in order to enhance and maximise peripheral perfusion. The study aimed to enhance peripheral perfusion and evaluate the primary clinical endpoint endothelial function, as well as to perform preliminary analysis of the ankle brachial index (ABI) and walking distance. RESULTS: Doppler flow measurements in the lower limb (ankle) validated that maximum blood flow velocity during systole and acceleration doubled during ISRT. Study results demonstrated that long-term ISRT significantly increased flow-mediated dilation (FMD) in the brachial artery (0.13+/- 0.09 mm to 0.38+/- 0.05 mm; p < 0.05), while nitromediated dilation (0.36+/- 0.10 mm to 0.45+/- 0.08 mm) remained and common femoral artery FMD did not reach statistical significance (0.38+/- 0.08 mm to 0.67+/- 0.19 mm; p<0.05). Initial claudication distance considerably improved for all patients after 30 hours of ISRT (92.6 +/- 8.2 metres to 280+/- 101.3 metres, p<0.05), just like the absolute claudication distance, which showed a more than 2.5-fold increase (167.8+/- 18.1 metres to 446.7+/- 133.3 metres; p<0.05). The ABI did not improve (0.58+/- 0.03 to 0.65+/- 0.04). CONCLUSIONS: Our data demonstrate that long-term ISRT is a potential novel non-invasive treatment to improve endothelial function and absolute pain-free walking distance for PAD patients.

Morphological evidence of a potential arrhythmogenic substrate in the caudal and cranial vena cava in horses.

BACKGROUND: Three-dimensional electro-anatomical mapping, previously performed in horses with atrial arrhythmias, has demonstrated the medial region of the caudal vena cava (CaVC), 1-8 cm caudal to the fossa ovalis, as an anatomical predilection site for atrial tachycardia associated with areas of slow conduction and conduction block. Slow conduction has also been recorded in the cranial vena cava (CrVC). OBJECTIVES: To investigate the morphological characteristics of the myocardial sleeves (MS) in the CaVC and CrVC, in order to identify a potential substrate of right sided atrial arrhythmias. STUDY DESIGN: Cross sectional. METHODS: Post-mortem dissection of 37 hearts from adult warmblood horses without known cardiovascular disease. Macroscopic examination of the myocardial distribution, evaluated the MS area, length, width, and shape in the CaVC and the CrVC. At least 2 samples from each vena cava MS were histologically examined using Masson's trichrome staining. RESULTS: Myocardial sleeves into the medial CaVC and into the CrVC were observed in all horses and showed variations in distribution, shape, and size between horses. Their mean ± standard deviation length from the limbus into the CaVC reached 5.7 ± 1.0 cm (maximum 8.3 cm), and from the azygos vein into the CrVC 5.3 ± 1.6 cm (maximum 8.6 cm). Myocardium-free islands were observed in the CaVC and CrVC in 30% and 6% of horses, respectively. Histologically, MS showed a non-uniform myocardial fibre arrangement, with presence of fibroadipose tissue, features known to result in slow conduction and pro-arrhythmia. MAIN LIMITATIONS: Study only included horses without history of atrial arrhythmia. CONCLUSIONS: Myocardial sleeves are present in both CaVC and CrVC, showing anatomical variations between horses. Tissue characteristics known to favour re-entry were identified indicating that these venae cavae MS are a potential substrate for atrial tachyarrhythmias and a target for treatment by ablation.

CONTEXTE: La modélisation électro-anatomique tridimensionnelle, réalisée auparavant chez des chevaux souffrant d'arythmies atriales, a démontré que la région médiale de la veine cave caudale (CaVC), 1-8 cm caudalement à la fossae ovalis, représente un site anatomique de prédilection pour la tachycardie auriculaire associée à des zones de conduction ralentie et des blocs de conduction. Une conduction ralentie a aussi été enregistrée dans la veine cave crâniale (CrVC). OBJECTIFS: Investiguer les caractéristiques morphologiques des manchons myocardiques (Myocardial sleeve;MS) dans les CAVC et CrVC, afin d'identifier un substrat potentiel d'arythmies atriales du côté droit du cœur. TYPE D'ÉTUDE: Étude transversale. MÉTHODES: Dissection post-mortem de 37 cœurs de chevaux à sang chaud adultes sans historique de maladie cardiovasculaire. Lors de l'examen macroscopique de la distribution myocardique, la surface des MS, leur longueur, largeur et forme ont été évaluées dans les CAVC et CrVC. Les MS ont été examinées microscopiquement dans au moins 2 échantillons de chaque veine cave, en utilisant la coloration de Masson Trichrome. RÉSULTATS: Les MS à l'aspect médial de la CaVC et à l'intérieur de la CrVC ont été inspectées chez tous les chevaux et ont montré des variations de distribution, forme et grandeur entre chevaux. Leur moyenne de longueur ± déviation standard du limbe de la fossae ovalis dans la CaVC était de 5.7 ± 1.0 cm (maximum 8.3 cm) et de la veine azygos dans la CrVC, de 5.3 ± 1.6 cm (maximum 8.6 cm). Des Îlots dénudés de myocarde ont été observé dans la CaVC et CrVC dans 30% et 6% des chevaux respectivement. À l'histologie, les MS ont montré des fibres myocardiques organisées de façon non-uniforme, avec présence de tissue fibroadipeux, caractéristiques connues pour entraîner une conduction lente et favoriser l'arythmie. LIMITES PRINCIPALES: Étude incluant seulement des chevaux sans historique d'arythmie atriale. CONCLUSION: Les MS sont présentes dans les CaVC et CrVC, démontrant les variations anatomiques entre chevaux. Des caractéristiques tissulaires connues pour favoriser une conduction ralentie ont été identifiés indiquant que les MS représentent un substrat potentiel des tachyarythmies atriales et une cible pour un traitement par ablation.

Effect of physiological and pharmacological stress on heart rate, blood pressure, and echocardiographic measurements in healthy Warmblood horses.

BACKGROUND: Echocardiographic measurements are important prognostic indicators but might be influenced by heart rate and blood pressure. This is particularly important when comparing repeated examinations. HYPOTHESIS: To determine the effect of physiological stress at mildly increased heart rates and pharmacological challenge using IV administration of N-butylscopolammonium bromide and metamizol sodium on heart rate, blood pressure, and echocardiographic measurements. ANIMALS: Twenty healthy Warmblood horses. METHODS: Randomized crossover study. Horses were examined echocardiographically by 2-dimensional, M-mode, pulsed wave (PW) Doppler, and PW tissue Doppler imaging with simultaneous ECG recording and noninvasive blood pressure measurements during rest, physiological stress, and pharmacological challenge. Cardiac dimensions and functions were measured by a blinded observer. Data were analyzed using repeated-measures analysis of variance. RESULTS: Mean heart rate and arterial blood pressure were significantly higher during physiological stress (46 ± 2 bpm, 93 ± 16 mm Hg) and pharmacological challenge (62 ± 13 bpm, 107 ± 17 mm Hg) compared with rest (34 ± 3 bpm, 86 ± 12 mm Hg; P < .05). Compared with rest, physiological stress resulted in increased left atrial fractional area change (34.3 ± 7.5 vs 27.3 ± 5.1%; P = .01) and left ventricular late diastolic radial wall motion velocity (13 ± 3 vs 10 ± 2 cm/s; P = .01) but had no significant effect on most other echocardiographic variables. Compared with rest, pharmacological challenge led to significantly decreased left atrial and diastolic ventricular dimensions (left ventricular internal diameter: 10.3 ± 0.9 vs 10.7 ± 0.8 cm; P = .01), increased aortic and pulmonary diameters, and ventricular wall thickness. CONCLUSIONS AND CLINICAL IMPORTANCE: Physiological stress at mildly increased heart rates significantly enhanced atrial pump function. Larger heart rate and blood pressure increases during pharmacological challenge resulted in altered cardiac dimensions. This should be taken into account when evaluating echocardiographic measurements at increased heart rates.

Feasibility of transthoracic echocardiographic guidance for multicatheter electrophysiological mapping studies in horses.

BACKGROUND: Improved characterization of arrhythmias is based on minimally invasive catheterizations. However, these catheterizations have been poorly explored in horses because apart from 3-dimensional (3D) mapping systems, continuous guidance of the catheter's position with adequate detail is difficult using current imaging modalities. HYPOTHESIS: Position multiple electrophysiology catheters simultaneously at predetermined strategical positions in the heart using transthoracic echocardiographic guidance. ANIMALS: Eight adult healthy horses. METHODS: Observational study. Two electrophysiological studies were performed: 1 procedure with catheters positioned in the right heart in the standing sedated horse and 1 procedure under general anesthesia with catheters positioned in the left heart. Except for the coronary sinus catheter, each catheter positioning was simultaneously guided by right-parasternal transthoracic echocardiography and 3D electro-anatomical mapping. RESULTS: For each catheter position, a central imaging plane was taken as the starting point, after which the imaging probe was shifted, rotated, and angulated to visualize the catheter over its entire length, including its distal electrode. Catheter positionings in the right heart and left ventricle were successfully guided in the majority of the horses whereas catheter positionings in the left atrium, and especially the pulmonary veins, were challenging to guide echocardiographically. CONCLUSIONS AND CLINICAL IMPORTANCE: Ultrasound guidance of catheters to specific positions useful for electrophysiological mapping was feasible in the right heart and left ventricle but challenging for the left atrium. This approach creates a perspective for minimally invasive arrhythmia diagnosis without the need for a 3D mapping system. Left parasternal views and intracardiac echocardiography might provide better guidance for left atrial positions.

Development of an atrial transseptal puncture procedure in horses to access the left heart: An ultrasound-guided jugular vein and transhepatic approach.

BACKGROUND: Radiofrequency ablation has been successfully applied to treat right atrial arrhythmias in horses. Ablation of left-sided arrhythmias requires a retrograde transarterial approach which is complicated. In human medicine, the left atrium is accessed through transseptal puncture (TSP) of the fossa ovalis (FO) using a caudal approach via the femoral vein. OBJECTIVES: To develop a zero fluoroscopy TSP technique for horses using a jugular vein (cranial) and transhepatic (caudal) approach. STUDY DESIGN: In vivo experimental study. METHODS: Transseptal puncture was performed in 18 horses admitted for euthanasia and donated for scientific research under general anaesthesia: using a jugular vein approach (10 horses), a transhepatic approach (2 horses) or both (6 horses). Radiofrequency energy was applied on a guidewire to perforate the FO and allow sheath advancement under intracardiac and transthoracic echocardiographic guidance. Puncture lesions were inspected post-mortem. RESULTS: Transseptal puncture was successful in 17/18 horses, of which 15/16 jugular vein approaches and 5/8 transhepatic approaches. Failure was due to technical malfunction, inability to advance the guidewire toward the heart and inability to advance the sheath through the FO. Intracardiac echocardiography was essential to safely guide the puncture process. Atrial arrhythmias caused by the TSP occurred in 13/18 horses. Puncture lesions were found in the right atrium in the FO region, and left atrium ventral to pulmonary vein ostium III. MAIN LIMITATIONS: Because in several horses two approaches were tested consecutively, it cannot be excluded that the second TSP was performed at the previous puncture site. Due to the developmental nature of the study the approaches were not randomised and did not allow comparison. CONCLUSION: Transseptal puncture is feasible in horses using ultrasound guidance and allows for electrophysiological exploration of the left heart. Further studies are needed to evaluate post-operative follow-up.

Intracardiac ultrasound-guided transseptal puncture in horses: Outcome, follow-up, and perioperative anticoagulant treatment.

BACKGROUND: Cardiac catheterizations in horses are mainly performed in the right heart, as access to the left heart traditionally requires an arterial approach. Transseptal puncture (TSP) has been adapted for horses but data on follow-up and closure of the iatrogenic atrial septal defect (iASD) are lacking. HYPOTHESIS/OBJECTIVES: To perform TSP and assess postoperative complications and iASD closure over a minimum of 4 weeks. ANIMALS: Eleven healthy adult horses. METHODS: Transseptal puncture was performed under general anesthesia. Serum cardiac troponin I concentrations were measured before and after puncture. Weekly, iASD closure was monitored using transthoracic and intracardiac echocardiography. Relationship between activated clotting time and anti-factor Xa activity during postoperative enoxaparin treatment was assessed in vitro and in vivo. RESULTS: Transseptal puncture was successfully achieved in all horses within a median duration of 22 (range, 10-104) minutes. Balloon dilatation of the puncture site for sheath advancement was needed in 4 horses. Atrial arrhythmias occurred in 9/11 horses, including atrial premature depolarizations (N = 1), atrial tachycardia (N = 5), and fibrillation (N = 3). Serum cardiac troponin I concentrations increased after TSP, but remained under the reference value in 10/11 horses. Median time to iASD closure was 14 (1-35) days. Activated clotting time correlated with anti-factor Xa activity in vitro but not in vivo. CONCLUSIONS AND CLINICAL IMPORTANCE: Transseptal puncture was successfully performed in all horses. The technique was safe and spontaneous iASD closure occurred in all horses. Clinical application of TSP will allow characterization and treatment of left-sided arrhythmias in horses.

Impact of the Timing of Mechanical Circulatory Support on the Outcomes in Myocardial Infarction-Related Cardiogenic Shock: Subanalysis of the PREPARE CS Registry.

(1) Background: Mechanical circulatory support (MCS) in myocardial infarction-associated cardiogenic shock is subject to debate. This analysis aims to elucidate the impact of MCS's timing on patient outcomes, based on data from the PREPARE CS registry. (2) Methods: The PREPARE CS prospective registry includes patients who experienced cardiogenic shock (SCAI classes C-E) and were subsequently referred for cardiac catheterization. Our present analysis included a subset of this registry, in whom MCS was used and who underwent coronary intervention due to myocardial infarction. Patients were categorized into an Upfront group and a Procedural group, depending on the timing of MCS's introduction in relation to their PCI. The endpoint was in-hospital mortality. (3) Results: In total, 71 patients were included. MCS was begun prior to PCI in 33 (46%) patients (Upfront), whereas 38 (54%) received MCS during or after the initiation of PCI (Procedural). The groups' baseline characteristics and hemodynamic parameters were comparable. The Upfront group had a higher utilization of the Impella® device compared to extracorporeal membrane oxygenation (67% vs. 33%), while the Procedural group exhibited a balanced use of both (50% vs. 50%). Most patients suffered from multi-vessel disease in both groups (82% vs. 84%, respectively; p = 0.99), and most patients required a complex PCI procedure; the latter was more prevalent in the Upfront group (94% vs. 71%, respectively; p = 0.02). Their rates of complete revascularization were comparable (52% vs. 34%, respectively; p = 0.16). Procedural CPR was significantly more frequent in the Procedural group (45% vs. 79%, p < 0.05); however, in-hospital mortality was similar (61% vs. 79%, respectively; p = 0.12). (4) Conclusions: The upfront implantation of MCS in myocardial infarction-associated CS did not provide an in-hospital survival benefit.

Tolerability safety and effectiveness of enhanced external counterpulsation versus individual shear rate therapy in patients with lower extremity atherosclerotic disease: A prospective pilot clinical trial.

AIMS: We aimed to investigate the tolerability, safety, and effectiveness of enhanced external counterpulsation therapy (EECP) versus individual shear rate therapy (ISRT) in patients with lower extremity atherosclerotic disease (LEAD). METHODS: Eighteen patients (age: 73.1 ± 6 years) underwent EECP and ISRT, each daily over five consecutive days in a cross-over design with a 1 week resting period in between the two regimens. A quality-of-life questionnaire was used to assess the therapy experience. Oxygen saturation (SO2 ), relative hemoglobin amount (rHb) and blood flow (Flow) in the capillary-venous-system (microcirculation) of the skin were monitored continuously during all therapy sessions using the micro-lightguide spectrophotometer, also known as oxygen to see (O2C). The effects of EECP and ISRT on the renal function and skeletal muscles were evaluated using serial blood and urine tests. RESULTS: EECP therapy had to be terminated early before the end of the 5th session in 10 patients (55.6%) because of discomfort. Four patients (22.2%) experienced signs of critical limb ischaemia under EECP. The total score of the quality-of-life questionnaire was significantly higher (= better tolerated) post-ISRT compared with EECP. Microcirculation monitoring revealed that ISRT significantly increased the SO2 , blood flow and rHb during the therapy. All three parameters remained significantly increased in the observation period after ISRT. The serum levels of creatin kinase and myoglobin increased significantly under EECP. CONCLUSIONS: ISRT significantly improves tolerability, safety, and effectiveness over EECP in patients with LEAD.

Three-dimensional electro-anatomical mapping and radiofrequency ablation as a novel treatment for atrioventricular accessory pathway in a horse: A case report.

We describe the diagnosis and treatment of an atrioventricular accessory pathway (AP) in a horse using 3-dimensional electro-anatomical mapping (3D EAM) and radiofrequency catheter ablation (RFCA). During routine evaluation of the horse, intermittent ventricular pre-excitation was identified on the ECG, characterized by a short PQ interval and abnormal QRS morphology. A right cranial location of the AP was suspected from the 12-lead ECG and vectorcardiography. After precise localization of the AP using 3D EAM, ablation was performed and AP conduction was eliminated. Immediately after recovery from anesthesia an occasional pre-excited complex still was observed, but a 24-hour ECG and an ECG during exercise 1 and 6 weeks after the procedure showed complete disappearance of pre-excitation. This case shows the feasibility of 3D EAM and RFCA to identify and treat an AP in horses.

Successful caudal vena cava and pulmonary vein isolation in healthy horses using 3D electro-anatomical mapping and a contact force-guided ablation system.

BACKGROUND: Recently, treatment of equine atrial tachycardia by three-dimensional electro-anatomical mapping (3D EAM) and radiofrequency catheter ablation (RFCA) has been described. Myocardial sleeves in the caudal vena cava and pulmonary veins are a potential trigger for initiation and perpetuation of atrial tachycardia and atrial fibrillation in the horse. Isolation of these myocardial sleeves by RFCA may be an effective treatment for these arrhythmias. OBJECTIVES: To describe the feasibility of 3D EAM and RFCA to isolate caudal vena cava and pulmonary veins in adult horses using 3D mapping and a contact force (CF)-guided ablation system. STUDY DESIGN: In vivo experiments. METHODS: 3D EAM and RFCA was performed in five horses without cardiovascular disease under general anaesthesia, using the CF-guided system CARTO®3. Point-by-point RFCA aimed for isolation of caudal vena cava and pulmonary veins. Radiofrequency energy was delivered in power-controlled mode with a target power of 45 W, CF between 10 and 15 g and 30 mL/min irrigation rate, until an ablation-index of 450-500 was reached. RESULTS: In the right atrium, myocardial sleeves of the caudal vena cava were isolated (n = 5). In the left atrium, isolation of ostium II (n = 3), ostium III (n = 1) and ostium I, II and III en bloc (n = 1) was performed. Successful isolation was confirmed by entrance and exit block. MAIN LIMITATIONS: Horses were euthanised at the end of the procedure, so long term effects such as potential reconnection of isolated veins could not be studied. CONCLUSIONS: This is the first description of 3D EAM and RFCA with CARTO®3 in horses, thereby showing the technical feasibility and successful caudal vena cava and pulmonary vein isolation. CF measurement allowed monitoring of catheter-tissue contact, resulting in efficient acute lesion creation as confirmed by entrance and exit block. This is a promising treatment for cardiac arrhythmias in horses.

Leukocyte telomere length and mitochondrial DNA copy number associate with endothelial function in aging-related cardiovascular disease.

Background: We investigated the association between leukocyte telomere length, mitochondrial DNA copy number, and endothelial function in patients with aging-related cardiovascular disease (CVD). Methods: In total 430 patients with CVD and healthy persons were enrolled in the current study. Peripheral blood was drawn by routine venipuncture procedure. Plasma and peripheral blood mononuclear cells (PBMCs) were collected. Cell-free genomic DNA (cfDNA) and leukocytic genomic DNA (leuDNA) were extracted from plasma and PBMCs, respectively. Relative telomere length (TL) and mitochondrial DNA copy number (mtDNA-CN) were analyzed using quantitative polymerase chain reaction. Endothelial function was evaluated by measuring flow-mediated dilation (FMD). The correlation between TL of cfDNA (cf-TL), mtDNA-CN of cfDNA (cf-mtDNA), TL of leuDNA (leu-TL), mtDNA-CN of leuDNA (leu-mtDNA), age, and FMD were analyzed based on Spearman's rank correlation. The association between cf-TL, cf-mtDNA, leu-TL, leu-mtDNA, age, gender, and FMD were explored using multiple linear regression analysis. Results: cf-TL positively correlated with cf-mtDNA (r = 0.1834, P = 0.0273), and leu-TL positively correlated with leu-mtDNA (r = 0.1244, P = 0.0109). In addition, both leu-TL (r = 0.1489, P = 0.0022) and leu-mtDNA (r = 0.1929, P < 0.0001) positively correlated with FMD. In a multiple linear regression analysis model, both leu-TL (ß = 0.229, P = 0.002) and leu-mtDNA (ß = 0.198, P = 0.008) were positively associated with FMD. In contrast, age was inversely associated with FMD (ß = -0.426, P < 0.0001). Conclusion: TL positively correlates mtDNA-CN in both cfDNA and leuDNA. leu-TL and leu-mtDNA can be regarded as novel biomarkers of endothelial dysfunction.

Angiotensin receptor-neprilysin inhibitor improves coronary collateral perfusion.

Background: We investigated the pleiotropic effects of an angiotensin receptor-neprilysin inhibitor (ARNi) on collateral-dependent myocardial perfusion in a rat model of coronary arteriogenesis, and performed comprehensive analyses to uncover the underlying molecular mechanisms. Methods: A rat model of coronary arteriogenesis was established by implanting an inflatable occluder on the left anterior descending coronary artery followed by a 7-day repetitive occlusion procedure (ROP). Coronary collateral perfusion was measured by using a myocardial particle infusion technique. The putative ARNi-induced pro-arteriogenic effects were further investigated and compared with an angiotensin-converting enzyme inhibitor (ACEi). Expression of the membrane receptors and key enzymes in the natriuretic peptide system (NPS), renin-angiotensin-aldosterone system (RAAS) and kallikrein-kinin system (KKS) were analyzed by quantitative polymerase chain reaction (qPCR) and immunoblot assay, respectively. Protein levels of pro-arteriogenic cytokines were measured by enzyme-linked immunosorbent assay, and mitochondrial DNA copy number was assessed by qPCR due to their roles in arteriogenesis. Furthermore, murine heart endothelial cells (MHEC5-T) were treated with a neprilysin inhibitor (NEPi) alone, or in combination with bradykinin receptor antagonists. MHEC5-T proliferation was analyzed by colorimetric assay. Results: The in vivo study showed that ARNis markedly improved coronary collateral perfusion, regulated the gene expression of KKS, and increased the concentrations of relevant pro-arteriogenic cytokines. The in vitro study demonstrated that NEPis significantly promoted MHEC5-T proliferation, which was diminished by bradykinin receptor antagonists. Conclusion: ARNis improve coronary collateral perfusion and exert pro-arteriogenic effects via the bradykinin receptor signaling pathway.

Angiotensin-converting enzyme inhibitors stimulate cerebral arteriogenesis.

AIM: Arteriogenesis constitutes the most efficient endogenous rescue mechanism in cases of cerebral ischaemia. The aim of this work was to investigate whether angiotensin-converting enzyme inhibitors (ACEi) stimulates, and angiotensin II receptor type 1 blockers (ARB) inhibits cerebral collateral growth by applying a three-vessel occlusion (3-VO) model in rat. METHODS: Cerebral collateral growth was measured post 3-VO (1) by assessing blood flow using the cerebrovascular reserve capacity (CVRC) technique, and (2) by assessing vessel diameters in the posterior cerebral artery (PCA) via the evaluation of latex angiographies. A stimulatory effect on arteriogenesis was investigated for ACEi administration ± bradykinin receptor 1 (B1R) and 2 (B2R) blockers, and an inhibitory effect was analysed for ARB administration. Results were validated by immunohistochemical analysis and mechanistic data were collected by human umbilical vein endothelial cell (HUVEC) viability or scratch assay and monocyte (THP-1) migration assay. RESULTS: An inhibitory effect of ARB on arteriogenesis could not be demonstrated. However, collateral growth measurements demonstrated a significantly increased CVRC and PCA diameters in the ACEi group. ACEi stimulates cell viability and migration, which could be partially reduced by additional administration of bradykinin receptor 1 inhibitor (B1Ri). ACEi inhibits the degradation of pro-arteriogenic bradykinin derivatives, but combined ACEi + B1Ri + B1Ri (BRB) treatment did not reverse the stimulatory effect. Yet, co-administration of ACEi + BRB enhances arteriogenesis and cell migration. CONCLUSION: We demonstrate a potent stimulatory effect of ACEi on cerebral arteriogenesis in rats, presumable via B1R. However, results imply a pleiotropic and compensatory effect of ACEi on bradykinin receptor-stimulated arteriogenesis.

Individual shear rate therapy (ISRT)-further development of external counterpulsation for decreasing blood pressure in patients with symptomatic coronary artery disease (CAD).

Individual shear rate therapy (ISRT) evolved from external counterpulsation with individual treatment pressures based on Doppler ultrasound measurements. In this study, we assessed the effect of ISRT on blood pressure (BP) in patients with coronary artery disease (CAD). Eighty-four patients with symptomatic CAD were included in the study. Forty-one patients were enrolled for 6 weeks, comprising 30 sessions of ISRT; 43 age- and sex-matched patients represented the control group. The 24-h BP was determined by measuring the pulse transit time before and after 6 weeks of ISRT or the time-matched control. Participants were divided into three groups according to the 24-h BP before treatment: BP1 < 130/80 mmHg (normotensive); BP2 ≥ 130-140/80 mmHg (moderate hypertensive); BP3 > 140/80 mmHg (hypertensive). After 30 sessions of ISRT, the 24-h BP decreased significantly, whereas no changes were observed in the controls. The BP-lowering effect correlated with the 24-h BP before therapy (systolic: r = -0.78; p < 0.001; diastolic: r = -0.76; p < 0.001). In BP1, the systolic BP decreased by 4.3 ± 6.4 mmHg (p = 0.011), and the diastolic BP decreased by 4.8 ± 11.0 mmHg (p = 0.032); in BP2, the systolic BP decreased by 13.3 ± 7.5 mmHg (p < 0.001), and the diastolic BP decreased by 5.0 ± 7.5 mmHg (p = 0.002); and in BP3, the systolic BP decreased by 22.9 ± 11.4 mmHg (p < 0.001), and the diastolic BP decreased by 9.1 ± 9.5 mmHg (p = 0.003). Our findings demonstrate that ISRT reduces BP in patients with CAD. The higher the initial BP the greater the lowering effect.

Supplemental Fibrinogen Restores Platelet Inhibitor-Induced Reduction in Thrombus Formation without Altering Platelet Function: An In Vitro Study.

BACKGROUND: For patients treated with dual antiplatelet therapy, standardized drug-specific 3-to-7 day cessation is recommended prior to major surgery to reach sufficient platelet function recovery. Here we investigated the hypothesis that supplemental fibrinogen might mitigate the inhibitory effects of antiplatelet therapy. METHODS AND RESULTS: To this end blood from healthy donors was treated in vitro with platelet inhibitors, and in vitro thrombus formation and platelet activation were assessed. Ticagrelor, acetylsalicylic acid, the combination of both, and tirofiban all markedly attenuated the formation of adherent thrombi, when whole blood was perfused through collagen-coated microchannels at physiological shear rates. Addition of fibrinogen restored in vitro thrombus formation in the presence of antiplatelet drugs and heparin. However, platelet activation, as investigated in assays of P-selectin expression and calcium flux, was not altered by fibrinogen supplementation. Most importantly, fibrinogen was able to restore in vitro thrombogenesis in patients on maintenance dual antiplatelet therapy after percutaneous coronary intervention. CONCLUSION: Thus, our in vitro data support the notion that supplementation of fibrinogen influences the perioperative hemostasis in patients undergoing surgery during antiplatelet therapy by promoting thrombogenesis without significantly interfering with platelet activation.