PFKFB3 gene deletion in endothelial cells inhibits intraplaque angiogenesis and lesion formation in a murine model of venous bypass grafting

Vein grafting is a frequently used surgical intervention for cardiac revascularization. However, vein grafts display regions with intraplaque (IP) angiogenesis, which promotes atherogenesis and formation of unstable plaques. Graft neovessels are mainly composed of endothelial cells (ECs) that largely depend on glycolysis for migration and proliferation. In the present study, we aimed to investigate whether loss of the glycolytic flux enzyme phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3) in ECs inhibits IP angiogenesis and as such prevents unstable plaque formation. To this end, apolipoprotein E deficient (ApoE(-/-)) mice were backcrossed to a previously generated PFKFB3(fl/fl) Cdh5(iCre) mouse strain. Animals were injected with either corn oil (ApoE(-/-)PFKFB3(fl/fl)) or tamoxifen (ApoE(-/-)PFKFB3(ECKO)), and were fed a western-type diet for 4 weeks prior to vein grafting. Hereafter, mice received a western diet for an additional 28 days and were then sacrificed for graft assessment. Size and thickness of vein graft lesions decreased by 35 and 32%, respectively, in ApoE(-/-)PFKFB3(ECKO) mice compared to controls, while stenosis diminished by 23%. Moreover, vein graft lesions in ApoE(-/-)PFKFB3(ECKO) mice showed a significant reduction in macrophage infiltration (29%), number of neovessels (62%), and hemorrhages (86%). EC-specific PFKFB3 deletion did not show obvious adverse effects or changes in general metabolism. Interestingly, RT-PCR showed an increased M2 macrophage signature in vein grafts from ApoE(-/-)PFKFB3(ECKO) mice. Altogether, EC-specific PFKFB3 gene deletion leads to a significant reduction in lesion size, IP angiogenesis, and hemorrhagic complications in vein grafts. This study demonstrates that inhibition of endothelial glycolysis is a promising therapeutic strategy to slow down plaque progression.Vascular Surger

INSPIRE: A European training network to foster research and training in cardiovascular safety pharmacology

Safety pharmacology is an essential part of drug development aiming to identify, evaluate and investigate undesirable pharmacodynamic properties of a drug primarily prior to clinical trials. In particular, cardiovascular adverse drug reactions (ADR) have halted many drug development programs. Safety pharmacology has successfully implemented a screening strategy to detect cardiovascular liabilities, but there is room for further refinement. In this setting, we present the INSPIRE project, a European Training Network in safety pharmacology for Early Stage Researchers (ESRs), funded by the European Commission's H2020-MSCA-ITN programme. INSPIRE has recruited 15 ESR fellows that will conduct an individual PhD-research project for a period of 36 months. INSPIRE aims to be complementary to ongoing research initiatives. With this as a goal, an inventory of collaborative research initiatives in safety pharmacology was created and the ESR projects have been designed to be complementary to this roadmap. Overall, INSPIRE aims to improve cardiovascular safety evaluation, either by investigating technological innovations or by adding mechanistic insight in emerging safety concerns, as observed in the field of cardio-oncology. Finally, in addition to its hands-on research pillar, INSPIRE will organize a number of summer schools and workshops that will be open to the wider community as well. In summary, INSPIRE aims to foster both research and training in safety pharmacology and hopes to inspire the future generation of safety scientists

Evaluation of drug related problems in Belgium: focus on corticosteroids

Background: Pharmacists have an important role to play in detecting and resolving drug-related problems (DRPs). Most studies have shown that DRPs have a negative impact on clinical results and quality of life as well as on health care costs. Corticosteroids are often implicated in DRPs, in particular because of their side effects, incorrect use of the inhalation device or lack of adherence to the prescribed regimen.Purpose: The purpose of this work is to identify causes of DRPs and interventions performed by pharmacists on corticosteroid-related problems and to distinguish between problems related to inhaled and general corticosteroids.Methods: During 5 days of their internship, 530 final year students of pharmaceutical sciences in six Belgian universities collected DRPs encountered in community pharmacies, as well as related interventions performed by pharmacists. The DRPs’ electronic registration was done through an adapted tool based on the classification of Pharmaceutical Care Network Europe (PCNE - v 6.2). This tool was validated by pharmacists and allowed to measure the frequency and nature of DRPs.Findings: Pharmacists detected 16 733 DRPs in total. 555 DRPs (3.3%) related to corticosteroids, of which 115 were inhaled corticosteroids. The most common causes of corticosteroid-related problems (55%) were administrative and logistical factors and fraud. More than a half of the technical causes were incomplete prescriptions. Concerning clinical causes, 28% related to drug/device-use problems for inhaled corticosteroids, with 88% related to incorrect use of the inhalation device. For general corticosteroids, the most common clinical causes were drug choice (37%), including medication interaction (58%) and inappropriate medication (contraindications, side effects: 14%). Pharmacists’ intervention was similar for inhaled and general corticosteroids. Pharmacists intervened with the patient orally in 38% of total interventions, and in writing in more than 14% of interventions. Pharmacists did not react in 14% (inhaled corticosteroids) and 16% (general corticosteroids) of corticosteroid-related problems. These non-interventions covered, for example, interactions and incomplete prescriptions.Conclusion: Several corticosteroid-related problems were detected and solved. However, pharmacists barely intervened for non-observance and drug interactions. The introduction of a structured interview between the patient and the pharmacist would enable the patient to be educated and informed about his disease and treatment. Therefore, pharmacists’ training is essential to performing these interviews. More randomized studies should be done in community pharmacies to evaluate the impact of these interviews on patients and on therapeutic adherence in real time.info:eu-repo/semantics/publishe

Analysis by fast-atom bombardment tandem mass spectrometry of phosphatidylcholine isolated from heart mitochondrial fractions: Evidence of incorporation of monohydroxylated fatty acyl moieties

Phosphatidylcholine (PC) is one of the main phospholipids present in mitochondrial membranes. According to current knowledge, the predominant fatty acyl moieties in this phospholipid are 16, 18, 20, or 22 carbon atoms long with chains that contain only carbon and hydrogen atoms. We have conducted a detailed analysis of the fatty acid substituents of the phospholipids present in mitochondrial fractions by using fast-atom bombardment tandem mass spectrometry. Six monohydroxylated C-16 and C-18 fatty acyl moieties were found in PC extracted from mitochondrial fractions of rat heart. The structure of one of these monohydroxylated fatty acids has been elucidated and corresponded to 12-hydroxy 9-octadecenoic acid. indications that concern the structure of the five other monohydroxylated fatty acids are presented. These monohydroxylated fatty acyl groups are preferentially associated in the PC molecule with C-18 and C-20 fatty acyl moieties. We present arguments to suggest that the formation of these compounds is probably not due to a free-radical initiated mechanism. The potential implication of these monohydroxylated fatty acids in several physiological functions is suggested by the fact that free hydroxylated fatty acids that are identical or closely related to those found in the mitochondrial fractions possess various biological activities