Endothelial Nitric Oxide Synthesis Research Articles

Effect of Kaempferol on Modulation of Vascular Contractility Mainly through PKC and CPI-17 Inactivation.

In this study, we investigated the efficacy of kaempferol (a flavonoid found in plants and plant-derived foods such as kale, beans, tea, spinach and broccoli) on vascular contractibility and aimed to clarify the detailed mechanism underlying the relaxation. Isometric contractions of divested muscles were stored and linked with western blot analysis which was carried out to estimate the phosphorylation of myosin phosphatase targeting subunit 1 (MYPT1) and phosphorylation-dependent inhibitory protein for myosin phosphatase (CPI-17) and to estimate the effect of kaempferol on the RhoA/ROCK/CPI-17 pathway. Kaempferol conspicuously impeded phorbol ester-, fluoride- and a thromboxane mimetic-derived contractions regardless of endothelial nitric oxide synthesis, indicating its direct effect on smooth muscles. It also conspicuously impeded the fluoride-derived elevation in phospho-MYPT1 rather than phospho-CPI-17 levels and phorbol 12,13-dibutyrate-derived increase in phospho-CPI-17 and phospho-ERK1/2 levels, suggesting the depression of PKC and MEK activities and subsequent phosphorylation of CPI-17 and ERK1/2. Taken together, these outcomes suggest that kaempferol-derived relaxation incorporates myosin phosphatase retrieval and calcium desensitization, which appear to be modulated by CPI-17 dephosphorylation mainly through PKC inactivation.

Gestational Hypoxia Impaired Endothelial Nitric Oxide Synthesis Via miR-155-5p/NADPH Oxidase/Reactive Oxygen Species Axis in Male Offspring Vessels.

Nitric oxide (NO) is the most important vasodilator secreted by vascular endothelial cells, and its abnormal synthesis is involved in the development of cardiovascular disease. The prenatal period is a critical time for development and largely determines lifelong vascular health in offspring. Given the high incidence and severity of gestational hypoxia in mid-late pregnancy, it is urgent to further explore whether it affects the long-term synthesis of NO in offspring vascular endothelial cells. Pregnant Sprague-Dawley rats were housed in a normoxic or hypoxic (10.5% O2) chamber from gestation days 10 to 20. The thoracic aortas of fetal and adult male offspring were isolated for experiments. Gestational hypoxia significantly reduces the NO-dependent vasodilation mediated by acetylcholine in both the fetal and adult offspring thoracic aorta rings. Meanwhile, acetylcholine-induced NO synthesis is impaired in vascular endothelial cells from hypoxic offspring thoracic aortas. We demonstrate that gestational hypoxic offspring exhibit a reduced endothelial NO synthesis capacity, primarily due to increased expression of NADPH oxidase 2 and enhanced reactive oxygen species. Additionally, gestational hypoxic offspring show elevated levels of miR-155-5p in vascular endothelial cells, which is associated with increased expression of NADPH oxidase 2 and reactive oxygen species generation, as well as impaired NO synthesis. The present study is the first to demonstrate that gestational hypoxia impairs endothelial NO synthesis via the miR-155-5p/NADPH oxidase 2/reactive oxygen species axis in offspring vessels. These novel findings indicate that the detrimental effects of gestational hypoxia on fetal vascular function can persist into adulthood, providing new insights into the development of vascular diseases.

(011) Using Pre-Operative Low Intensity Vestibular Shockwave Therapy Reduces Opioid Use Post-Vestibulectomy

Abstract Introduction Surgeons performing complete vestibulectomy for neuro-proliferative vestibulodynia need to consider two competing post-operative interests: accomplishing acute pain relief in the early postoperative period; reducing risks of persistent post-operative opioid use. Data support that measures to decrease overall post-operative opioid use will decrease future opioid-related adverse events including opioid abuse and addiction. Sexual medicine providers are thus motivated to identify strategies such as pre-operative low intensity shockwave therapy (LISWT) to decrease opioid use in these vestibulectomy patients. To date, only one study has been published assessing the effect of pre-operative LISWT on surgical outcome. Dumfarth at al reported in a prospective randomized study that pre-operative LISWT significantly improved lower extremity wound healing compared to controls in patients undergoing vein harvesting for coronary artery bypass graft surgery. While Dumfarth et al did not directly assess post-operative pain, pre-operative LISWT was associated with statistically significantly increased satisfactory healing, reduced disturbance of healing and reduced minor skin infections, all consistent with less post-operative pain. It was hypothesized that pre-operative LISWT improved wound healing via enhanced blood supply and angiogenesis through expression of vascular endothelial growth factor (VEGF) and nitric oxide synthesis, and via anti-inflammatory action downregulating necrosis factor B activation thereby lowering inflammatory cytokines. Objectives We wished to see if pre-operative LISWT would reduce opioid use in women undergoing complete vestibulectomy. The objective of this study was to compare the use of opioids following vestibulectomy when LISWT with Urogold 100™ MTS was applied just prior to surgery versus no shockwave. LISWT is a non-invasive, non-pharmacologic, non-surgical treatment strategy with low morbidity that is FDA-cleared for pain amelioration in the US as non-significant risk in humans, Urogold 100™MTS provides unfocused electrohydraulic shockwaves with a unique parabolic reflector. Methods Patients scheduled for surgery were required to present to the hospital 4 days before surgery for a COVID-19 test. On that day and each subsequent day until surgery, based on the patient’s availability, the patient presented to the office for approximately 2400 shocks to the vestibule. This is a descriptive study of the patients’ post-operative experiences with pain. Results To date 4 women have undergone LISWT just prior to surgery. At the time of surgery Exparel was delivered to the site, allowing them 24-36 hours of anesthesia to the area so they could travel home easily. Patients were prescribed Percocet to be take every 4 hours as needed. Post-op patients routinely use this opioid 4-6 times a day for multiple days, causing constipation among the many side effects. The women who underwent LISWT immediately prior to surgery have experienced decreased levels of pain post-operatively such that they are using ½ tab of Percocet at bedtime or twice a day, thus decreasing opioid use with all of its potential side effects. Conclusions Performing LISWT using Urogold 100™MTS may reduce the need for opioid medications after total vestibulectomy with vaginal advancement flap as it may mediate pain after the surgical procedure. Disclosure No.

Nitric oxide metabolites in heart failure patients with reduced or preserved ejection fraction and effects of different exercise modes

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): German Centre for Cardiovascular Research (DZHK). Background Alterations of endothelial function and nitric oxide (NO) release are involved in pathophysiology and clinical symptoms of heart failure (HF). Regular exercise training improves vascular and endothelial function and is closely linked to mechanisms of NO pathway by enhancing endothelial NO synthesis. However, it is not well investigated how exercise training affects metabolites of the NO pathway and whether different exercise modes have different effects on NO bioavailability in HF. Purpose The aims of this analysis were to 1) investigate differences in serum levels of the NO metabolites L-arginine, homoarginine, asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) between patients with HF with reduced (HFrEF) or preserved ejection fraction (HFpEF) and 2) to investigate effects of high-intensity interval training (HIIT) and moderate continuous training (MCT) on these parameters. Methods Patients with HF from two clinical exercise intervention trials were included in this analysis. In both trials, patients were randomized (1:1:1) either to HIIT, MCT or guideline control (CG). Clinical data and blood samples were collected at baseline, after three months of supervised exercise training and after 12 months follow-up. NO metabolites were analysed from EDTA-plasma using liquid chromatography-tandem mass spectrometry (LC–MS/MS). Results Data from 366 patients with HF, including 206 HFrEF (median [1. and 3. quartile]: 61 [53;70] years, 18.9% females) and 160 HFpEF patients (71 [66;76] years, 65.6% females), was available for analysis. Patients with HFrEF or HFpEF showed no significant differences in NYHA classification (NYHA III: 30.6% vs. 26.2%, p=0.43) or presence of diabetes mellitus (23.8% vs. 26.2%, p=0.50), while HFpEF had a higher prevalence of hypertension (86.9% vs. 38.5%, p<0.001) but a smaller number of current smoker than HFrEF (4.4% vs. 18.0%, p<0.001). PeakVO2 (18.4 [14.5;22.6] ml/min/kg vs. 17.0 [14.3;20.4] ml/min/kg, p=0.03) and HDL-cholesterol (56 [46;65] mg/dl vs. 42 [36;54] mg/dl, p<0.001) were higher in HFpEF than in HFrEF. At baseline, homoarginine and ADMA levels were higher in HFrEF than in HFpEF, but no differences in L-arginine or SDMA could be observed (Table 1). Changes in metabolite levels from baseline to 3 and 12 months were not significantly different between groups (HIIT, MCT, CG) in either HF phenotype (all p>0.05). In all exercise groups, median adherence to training sessions was at 85% or above with no significant differences between groups (p>0.05). Conclusion At baseline, patients with HFrEF had significantly higher homoarginine and ADMA levels compared to patients with HFpEF. However, in both HF phenotypes neither MCT nor HIIT induced significant changes in serum levels of NO metabolites when compared to patients randomized to CG.

Specific dilation pattern in placental circulation and the NO/sGC role in preeclampsia placental vessels.

Endothelial functions in controlling blood flow in placental circulation are still unclear. The present study compares vascular dilations between placental circulation and other vessels, as well as between normal and preeclampsia placental vessels. Placental, umbilical, and other vessels (cerebral and mesenteric arteries) were collected from humans, sheep, and rats. Vasodilation was tested by JZ101 and DMT. Q-PCR, Western blot, and Elisa were used for molecular experiments. Endothelium-dependent/derived vasodilators, including acetylcholine, bradykinin, prostacyclin, and histamine, mediated no or minimal dilation in placental circulation, which was different from that in other vessels in sheep and rats. There were lower mRNA expressions of muscarinic receptors, histamine receptors, bradykinin receptor 2, endothelial nitric oxide synthesis (eNOS), and less nitric oxide (NO) in human umbilical vessels when compared with placental vessels. Exogenous NO donors (sodium nitroprusside, SNP) and soluble guanylate cyclase (sGC) activators (Bay41-2272) decreased the baseline of vessel tone in placental circulation in humans, sheep, and rats, but not in other arteries. The sGC inhibitor ODQ suppressed the reduced baseline caused by the SNP. The decreased baseline by SNP or Bay41-2272 was higher in placental vessels than in umbilical vessels, suggesting that the role of NO/sGC is more important in the placenta. NO concentrations in preeclampsia placental vessels were lower than those in control, while no significant change was found in umbilical plasma between the two groups. eNOS expression was similar between normal and preeclampsia placental vessels, but phosphorylated eNOS levels were significantly lower in preeclampsia. Following serotonin, SNP or Bay41-2272-mediated dilations were weaker in preeclampsia placental vessels. The decreased amplitude of SNP- or Bay41-2272 at baseline was smaller in preeclampsia. The decreased amplitudes of ODQ + SNP were comparable between the two groups. Despite higher beta sGC expression, sGC activity in the preeclampsia placenta was lower. This study demonstrated that receptor-mediated endothelium-dependent dilation in placental circulation was significantly weaker than other vessels in various species. The results, showed firstly, that exogenous NO played a role in regulating the baseline tone of placental circulation via sGC. Lower NO production and decreased NO/sGC could be one of the reasons for preeclampsia. The findings contribute to understanding specific features of placental circulation and provide information about preeclampsia in placental vessels.

Vascular Endothelial Dysfunctions with Hyperlipoproteinemia

Background: Cardiovascular diseases and, primarily coronary heart disease, remain among the leading causes of disability and mortality worldwide. Аim: To study the relationship between vascular endothelial dysfunction and hyperlipoproteinemia in experimental atherosclerosis. Materials and Methods: The experiments were carried out on 28 Chinchilla rabbits with an average weight of 2.5-3.0 kg. The action of drugs was studied in dynamics: the initial 3-month condition and after one month of drug administration. The results obtained were compared with those of the control and intact groups. Results: Long-term administration of cholesterol is accompanied by the development of hypertriglyceridemia, the severity of which depends on the duration of the experiment. Conclusion: An important role in the development of endothelial dysfunction in hypercholesterolemia is played by a decrease in the synthesis of endothelial nitric oxide and an increase in its active radicals, causing modification of low-density lipoproteins and their deposition in the vascular endothelium.

Aldosterone Synthase Inhibitors and Dietary Interventions: A Combined Novel Approach for Prevention and Treatment of Cardiovascular Disease.

Systemic hypertension (HTN) is the hallmark of cardiovascular disease and the forerunner of heart failure. These associations have been established over decades of research on essential HTN. Advancements in the treatment of patients diagnosed with HTN, consisting of alpha- or beta-adrenergic receptor blockers, calcium channel blockers, angiotensin-converting enzyme inhibitors, thiazide, or aldosterone receptor blockers known as anti-mineralocorticoids, in the presence or absence of low sodium salt diets, often fail to control blood pressure adequately to prevent morbidity and mortality. Low sodium diets have had limited success in controlling HTN because low sodium intake is associated with renin-angiotensin-aldosterone system upregulation. Therefore, upregulating aldosterone secretion, sodium, and water retention which, in turn, moves the blood pressure back toward the range of HTN dictated by the baroreceptor reset value, as a compensatory mechanism, especially in resistant HTN. These impediments to blood pressure control in HTN may have been effectively circumvented by the advent of a new class of drugs known as aldosterone synthase inhibitors, represented by baxdrostat.The mechanism of action of baxdrostat as an aldosterone synthase inhibitor demonstrates the inextricable linkage between sodium and blood pressure regulation. Theoretically, combining a low sodium diet with the activity of this aldosterone synthesis inhibitor should alleviate the adverse effect of renin-angiotensin-aldosterone system upregulation. Aldosterone synthesis inhibition should also decrease the oxidative stress and endothelial dysfunction associated with HTN, causing more endothelial nitric oxide synthesis, release, and vasorelaxation. To the best of our knowledge, this is the first systematic review to summarize evidence-based articles relevant to the use of a novel drug (aldosterone synthase inhibitor) in the treatment of HTN and cardiovascular disease. Making the current database of relevant information on baxdrostat and other aldosterone synthase inhibitors readily available will, no doubt, aid physicians and other medical practitioners in their decision-making about employing aldosterone synthase inhibitors in the treatment of patients.

The Relationship Between Coronary Collateral Circulation and Serum Adropin Levels.

Objective Coronary collateral circulation (CCC) are vascular structures that limit the infarct area, protect left ventricular function, and reduce the frequency of arrhythmia and mortality during myocardial ischemia and infarction. In this study, we examined the relationship between the development of CCC and serum adropin levels, which has been shown in previous studies to regulate endothelial functions and increase endothelial nitric oxide synthesis,in patients with acute myocardial infarction. Methods This study included 41 patients with insufficient CCC and 43 patients with well-developed CCC who were hospitalized for acute myocardial infarction and underwent coronary angiography. The Cohen-Rentrop classification was used to grade the CCC. The patients were divided into two groups according toRentrop grades: those with a 0-1 stage were considered as insufficientand those withgrades of 2-3 were considered as well-developed CCC. We took blood samples to measure the adropin levels within the first 24 hours of hospitalization. Results The mean age was 59.1±11.9 yearsand62 (73.8%)were male. The right coronary artery was the most frequently target vessel (n: 51, 60.7%), and the majority of the patients presented with ST-segment elevation myocardial infarction (STEMI) (n:58, 69%). The median interval between the severe chest pain and the intervention was significantly higher in patients with well-developed CCC (p=0.042). The serum adropin levels in patients with insufficient CCC were significantly lower than in those with well-developed CCC(196.3 [131.5 - 837.0] pg/mL vs. 235.5 [171.9 - 1124.2] pg/mL, p<0.001). Logistic regression analysis revealed that the circumflex artery as the target vessel, NSTEMI (non-STEMI) as the type of myocardial infarction, and serum adropin level were the independent risk factors for the prediction of poor coronary collateral vessel formation (p<0.05). Conclusions In this study, we found that in patients with acute myocardial infarction, those with well-developed CCC had higheradropin levels.

Tissue Derivation and Biological Sex Uniquely Mediate Endothelial Cell Protein Expression, Redox Status, and Nitric Oxide Synthesis.

Human endothelial cells are routinely utilized in cardiovascular research to provide a translational foundation for understanding how the vascular endothelium functions in vivo. However, little attention has been given to whether there are sex specific responses in vitro. Similarly, it is unclear whether endothelial cells derived from distinct tissues behave in a homogenous manner. Herein, we demonstrate that marked sex differences exist within, and between, commonly utilized human primary endothelial cells from healthy donors, with respect to redox status, nitric oxide synthesis, and associated proteins that can mediate their expression. Further, we demonstrate that endothelial cells respond uniquely to inflammatory insult in a sex- and tissue origin-dependent manner. Our findings suggest sex and tissue derivation may need to be considered when studying endothelial cells in vitro as cells derived from distinct tissue and sexes may not behave interchangeably.

The Effect of Luteolin on the Modulation of Vascular Contractility via ROCK and CPI-17 Inactivation.

In this investigation, we made a study of the efficacy of luteolin (a flavonoid found in plants such as vegetables, herbs and fruits) on vascular contractibility and to elucidate the mechanism underlying the relaxation. Isometric contractions of denuded muscles were stored and combined with western blot analysis which was conducted to assess the phosphorylation of myosin phosphatase targeting subunit 1 (MYPT1) and phosphorylation-dependent inhibitory protein for myosin phosphatase (CPI-17) and to examine the effect of luteolin on the RhoA/ROCK/CPI-17 pathway. Luteolin significantly alleviated phorbol ester-, fluoride- and thromboxane mimetic-elicited contractions regardless of endothelial nitric oxide synthesis, implying its direct effect on smooth muscle. It also significantly alleviated the fluoride-elicited elevation in pCPI-17 and pMYPT1 levels and phorbol 12,13-dibutyrate-elicited increase in pERK1/2 level, suggesting depression of ROCK and PKC/MEK activity and ensuing phosphorylation of MYPT1, CPI-17 and ERK1/2. Taken together, these results suggest that luteolin-elicited relaxation includes myosin phosphatase reactivation and calcium desensitization, which seems to be arbitrated by CPI-17 dephosphorylation via ROCK/PKC inhibition.

A Theoretical Model of the Mechanism by which Dietary Lipid Variance and Increased Oxysterols Allow Increased Formation of Soluble Endoglin in Preeclampsia

Endoglin (TGF‐β receptor III), an accessory receptor necessary for transforming growth factor‐β (TGF‐β) signaling, is essential during angiogenesis. Cleavage of endoglin by matrix metalloproteinase‐14 (MMP‐14) creates a soluble receptor (sEng) which is elevated in the sera of people with preeclampsia. Similarly to sflts which bind VEGF, sEng binds TGF‐β and impairs functions ranging from endothelial nitric oxide synthesis, vasodilation, and modulation of vascular permeability, to regulation of immune homeostasis, trophoblastic invasion and differentiation.Cleavage of endoglin to form sEng is upregulated in the presence of oxysterols. Initially, oxysterols were believed to upregulate MMP‐14 and down regulate its inhibiter TIMP3. However, in placental explants, expression MMP14 and TIMP3 mRNA were not altered by oxysterols and pravastatin did not decrease oxysterol‐induced upregulation of sEng. Membrane raft domains negatively regulate TGF‐β signaling, and aspirin, which disperses membrane rafts, induces TGF‐β signaling.This perspective considers the known changes in dietary lipids over the last 100 years, the possible effects of these changes on membrane raft formation, oxidative stress, and oxysterol formation, as well as the known effects of oxysterols on membrane thickness and fluidity and proposes an alternate mechanism by which oxysterols might increase sEng formation.

Pre-pperative low intensity vestibular shockwave therapy (LiSWT) decreases opioid use post-vestibulectomy: A pilot study

ABSTRACT Introduction Surgeons performing complete vestibulectomy with vaginal advancement flap reconstruction for suspected neuroproliferative vestibulodynia (NPV) take into consideration the need for acute pain relief in the early postoperative period as well as risks for persistent post-operative opioid use. Post-operative acute pain has traditionally been managed with narcotics which are effective but are highly addictive and can cause unwanted side effects including nausea/vomiting, impaired mental function, and severe constipation/obstipation. We serendipitously observed that one patient who tried to avoid surgery by undergoing numerous vestibular low intensity shockwave therapy (LiWST) treatments used very few narcotics post vestibulectomy. LiWST is FDA-cleared for pain amelioration. It has been hypothesized that LiSWT downregulates inflammatory cytokines, promotes enhancement of blood flow and angiogenesis through expression of vascular endothelial growth factor and nitric oxide synthesis. Objective We performed a pilot study using pre-operative LiSWT to determine whether or not it might decrease post-operative opioid use (oxycodone/acetaminophen tablets; 5mg/325mg) for acute pain management in patients undergoing complete vestibulectomy with vaginal advancement flap for suspected NPV. Method We studied the post-op opioid use of women who underwent complete vestibulectomy at our facility during the period January 1, 2019 through June 30, 2021. We assessed opioid use based on the patient's recall of the total oxycodone/acetaminophen tablets required in the first 30 days post-vestibulectomy. When possible, patients confirmed the use by counting tablets left in the pill container. Women scheduled to undergo vestibulectomy for suspected NPV were placed in the LiSWT study group if they agreed to undergo vestibular LiSWT treatment daily for the three business days immediately pre-operatively. Using the Urogold 100™MTS, LiSWT involved delivery of 1800 shocks to the vestibule per session, energy flux density 0.05-0.1 mJ/mm2, 3 Hz, membrane pressure 3. We compared the post-op opioid use by women in the LiSWT study group to the use by women who did not undergo pre-op LiSWT. Results Forty-five women (mean age 28 years; range 17 - 57) underwent complete vestibulectomy for suspected NPV during the 30 month period. Twenty women (mean age 27 +/- 9 years) agreed to undergo pre-op LiSWT and post-op opioid use was obtained in all. Twenty-five women (mean age 29 +/- 11 years) did not undergo pre-op LiSWT and opioid use was obtained in 19/25 (76%). Mean post-operative opioid use was 47 +/- 13 (range 25 - 90) tablets in those vestibulectomy patients who did not undergo pre-op vestibular LiSWT; opioid side effects included constipation, nausea, fainting, drowsiness, fatigue, and blurred vision. Mean post-op opioid use was 22 +/- 10 (range 1 - 26) tablets in the 20 vestibulectomy patients who did undergo pre-op vestibular LISWT sessions; opioid side effects were restricted to constipation, sleepiness, and nausea. Conclusion In this pilot study, pre-operative low intensity shockwave therapy decreased post-operative opioid use after complete vestibulectomy with vaginal advancement flap. While encouraging, more research is needed. Disclosure Work supported by industry: no.

A Pilot Study of Pre-Operative Vestibular Low Intensity Shockwave Therapy to Decrease Opioid Use Post-Vestibulectomy

ABSTRACT Introduction Surgeons performing complete vestibulectomy with vaginal advancement flap reconstruction for suspected neuroproliferative vestibulodynia (NPV) take into consideration the need for acute pain relief in the early postoperative period as well as risks for persistent post-operative opioid use. Post-operative acute pain has traditionally been managed with narcotics which are effective but are highly addictive and can cause unwanted side effects including nausea/vomiting, impaired mental function, and severe constipation/obstipation. We serendipitously observed that one patient who tried to avoid surgery by undergoing numerous vestibular low intensity shockwave therapy (LiWST) treatments used very few narcotics post vestibulectomy. LiWST is FDA-cleared for pain amelioration. It has been hypothesized that LiSWT downregulates inflammatory cytokines, promotes enhancement of blood flow and angiogenesis through expression of vascular endothelial growth factor and nitric oxide synthesis. Objective We performed a pilot study using pre-operative LiSWT to determine whether or not it might decrease post-operative opioid use (oxycodone/acetaminophen tablets; 5mg/325mg) for acute pain management in patients undergoing complete vestibulectomy with vaginal advancement flap for suspected NPV. Methods We studied the post-op opioid use of women who underwent complete vestibulectomy at our facility during the period January 1, 2019 through June 30, 2021. We assessed opioid use based on the patient's recall of the total oxycodone/acetaminophen tablets required in the first 30 days post-vestibulectomy. When possible, patients confirmed the use by counting tablets left in the pill container. Women scheduled to undergo vestibulectomy for suspected NPV were placed in the LiSWT study group if they agreed to undergo vestibular LiSWT treatment daily for the three business days immediately pre-operatively. Using the Urogold 100™MTS, LiSWT involved delivery of 1800 shocks to the vestibule per session, energy flux density 0.05-0.1 mJ/mm2, 3 Hz, membrane pressure 3. We compared the post-op opioid use by women in the LiSWT study group to the use by women who did not undergo pre-op LiSWT. Results Forty-five women (mean age 28 years; range 17 - 57) underwent complete vestibulectomy for suspected NPV during the 30 month period. Twenty women (mean age 27 +/- 9 years) agreed to undergo pre-op LiSWT and post-op opioid use was obtained in all. Twenty-five women (mean age 29 +/- 11 years) did not undergo pre-op LiSWT and opioid use was obtained in 19/25 (76%). Mean post-operative opioid use was 47 +/- 13 (range 25 - 90) tablets in those vestibulectomy patients who did not undergo pre-op vestibular LiSWT; opioid side effects included constipation, nausea, fainting, drowsiness, fatigue, and blurred vision. Mean post-op opioid use was 22 +/- 10 (range 1 - 26) tablets in the 20 vestibulectomy patients who did undergo pre-op vestibular LISWT sessions; opioid side effects were restricted to constipation, sleepiness, and nausea. Conclusions In this pilot study, pre-operative low intensity shockwave therapy decreased post-operative opioid use after complete vestibulectomy with vaginal advancement flap. While encouraging, more research is needed. Disclosure No

Surface Coating of Polyurethane Films with Gelatin, Aspirin and Heparin to Increase the Hemocompatibility of Artificial Vascular Grafts.

Purpose: A hemocompatible substrate can offer a wonderful facility for nitric oxide (NO) production by vascular endothelial cells in reaction to the inflammation following injuries. NO inhibits platelet aggregation this is especially critical in small-diameter vessels. Methods: The substrate films were made of polyurethane (PU) in a casting process and after plasma treatments, their surface was chemically decorated with polyethylene glycol (PEG) 2000, gelatin, gelatin-aspirin, gelatin-heparin and gelatin-aspirin-heparin. The concentrations of these ingredients were optimized in order to achieve the biocompatible values and the resulting modifications were characterized by water contact angle and Fourier transform infra-red (FTIR) assays. The values of NO production and platelet adhesion were then examined. Results: The water contact angle of the modified surface was reduced to 26±4∘ and the newly developed hydrophilic chemical groups were confirmed by FTIR. The respective concentrations of 0.05 mg/ml and 100 mg/mL were found to be the IC50 values for aspirin and heparin. However, after the surface modification with aspirin, the bioactivity of the substrate increased in compared to the other experimental groups. In addition, there was a synergistic effect between these reagents for NO synthesis. While, heparin inhibited platelet adhesion more than aspirin. Conclusion: Because of the highly hydrophilic nature of heparin, this reagent was hydrolyzed faster than aspirin and therefore its influence on platelet aggregation and cell growth was greater. Taken together, the results give the biocompatible concentrations of both biomolecules that are required for endothelial cell proliferation, NO synthesis and platelet adhesion.

Hyperuricemia-induced endothelial insulin resistance: the nitric oxide connection.

Hyperuricemia, defined as elevated serum concentrations of uric acid (UA) above 416µmol L-1, is related to the development of cardiometabolic disorders, probably via induction of endothelial dysfunction. Hyperuricemia causes endothelial dysfunction via induction of cell apoptosis, oxidative stress, and inflammation; however, it's interfering with insulin signaling and decreased endothelial nitric oxide (NO) availability, resulting in the development of endothelial insulin resistance, which seems to be a major underlying mechanism for hyperuricemia-induced endothelial dysfunction. Here, we elaborate on how hyperuricemia induces endothelial insulin resistance through the disruption of insulin-stimulated endothelial NO synthesis. High UA concentrations decrease insulin-induced NO synthesis within the endothelial cells by interfering with insulin signaling at either the receptor or post-receptor levels (i.e., proximal and distal steps). At the proximal post-receptor level, UA impairs the function of the insulin receptor substrate (IRS) and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) in the insulin signaling pathway. At the distal level, high UA concentrations impair endothelial NO synthase (eNOS)-NO system by decreasing eNOS expression and activity as well as by direct inactivation of NO. Clinically, UA-induced endothelial insulin resistance is translated into impaired endothelial function, impaired NO-dependent vasodilation, and the development of systemic insulin resistance. UA-lowering drugs may improve endothelial function in subjects with hyperuricemia.

The Effect of Galangin on the Regulation of Vascular Contractility via the Holoenzyme Reactivation Suppressing ROCK/CPI-17 rather than PKC/CPI-17.

In this study, we investigated the influence of galangin on vascular contractibility and to determine the mechanism underlying the relaxation. Isometric contractions of denuded aortic muscles were recorded and combined with western blot analysis which was performed to measure the phosphorylation of phosphorylation-dependent inhibitory protein of myosin phosphatase (CPI-17) and myosin phosphatase targeting subunit 1 (MYPT1) and to evaluate the effect of galangin on the RhoA/ROCK/CPI-17 pathway. Galangin significantly inhibited phorbol ester-, fluoride- and thromboxane mimetic-induced vasoconstrictions regardless of endothelial nitric oxide synthesis, suggesting its direct effect on vascular smooth muscle. Galangin significantly inhibited the fluoride-dependent increase in pMYPT1 and pCPI-17 levels and phorbol 12,13-dibutyrate-dependent increase in pERK1/2 level, suggesting repression of ROCK and MEK activity and subsequent phosphorylation of MYPT1, CPI-17 and ERK1/2. Taken together, these results suggest that galangin-induced relaxation involves myosin phosphatase reactivation and calcium desensitization, which appears to be mediated by CPI-17 dephosphorylation via not PKC but ROCK inactivation.

Novel insights for the role of nitric oxide in placental vascular function during and beyond pregnancy.

More than 30 years have passed since endothelial nitric oxide synthesis was described using the umbilical artery and vein endothelium. That seminal report set the cornerstone for unveiling the molecular aspects of endothelial function. In parallel, the understanding of placental physiology has gained growing interest, due to its crucial role in intrauterine development, with considerable long-term health consequences. This review discusses the evidence for nitric oxide (NO) as a critical player of placental development and function, with a special focus on endothelial nitric oxide synthase (eNOS) vascular effects. Also, the regulation of eNOS-dependent vascular responses in normal pregnancy and pregnancy-related diseases and their impact on prenatal and postnatal vascular health are discussed. Recent and compelling evidence has reinforced that eNOS regulation results from a complex network of processes, with novel data concerning mechanisms such as mechano-sensing, epigenetic, posttranslational modifications, and the expression of NO- and l-arginine-related pathways. In this regard, most of these mechanisms are expressed in an arterial-venous-specific manner and reflect traits of the fetal systemic circulation. Several studies using umbilical endothelial cells are not aimed to understand placental function but general endothelial function, reinforcing the influence of the placenta on general knowledge in physiology.

Melissa officinalis L. as a Nutritional Strategy for Cardioprotection.

This review aimed to provide a summary on the traditional uses, phytochemistry, and pharmacological activities in the cardiovascular system and cardiotoxicity of Melissa officinalis (MO), with the special emphasis on the protective mechanisms in different cardiovascular pathologies. MO is a perennial aromatic herb commonly known as lemon balm, honey balm, or bee balm, which belongs to Lamiaceae family. Active components are mainly located in the leaves or essential oil and include volatile compounds, terpenoid (monoterpenes, sesquiterpenes, triterpenes), and polyphenolic compounds [rosmarinic acid (RA), caffeic acid, protocatechuic acid, quercitrin, rhamnocitrin, luteolin]. For centuries, MO has been traditionally used as a remedy for memory, cognition, anxiety, depression, and heart palpitations. Up until now, several beneficial cardiovascular effects of MO, in the form of extracts (aqueous, alcoholic, and hydroalcoholic), essential oil, and isolated compounds, have been confirmed in preclinical animal studies, such as antiarrhythmogenic, negative chronotropic and dromotropic, hypotensive, vasorelaxant, and infarct size–reducing effects. Nonetheless, MO effects on heart palpitations are the only ones confirmed in human subjects. The main mechanisms proposed for the cardiovascular effects of this plant are antioxidant free radical–scavenging properties of MO polyphenols, amelioration of oxidative stress, anti-inflammatory effects, activation of M2 and antagonism of β1 receptors in the heart, blockage of voltage-dependent Ca2+ channels, stimulation of endothelial nitric oxide synthesis, prevention of fibrotic changes, etc. Additionally, the main active ingredient of MO-RA, per se, has shown substantial cardiovascular effects. Because of the vastness of encouraging data from animal studies, this plant, as well as the main ingredient RA, should be considered and investigated further as a tool for cardioprotection and adjuvant therapy in patients suffering from cardiovascular diseases.

The effect of α-lipoic acid treatment on plasma asymmetric dimethylarginine, a biomarker of endothelial dysfunction in diabetic neuropathy

IntroductionDiabetic neuropathy may develop on a background of hyperglycaemia and is associated with increased oxidative stress. Elevated asymmetric dimethylarginine (ADMA) levels are linked to oxidative stress reducing the synthesis of nitric oxide (NO) by uncoupling NO synthase. Oxidative stress induces considerable changes in nerve conduction velocity in diabetic patients. There is strong evidence that α-lipoic acid (ALA) as an antioxidant may improve nerve conduction and relieve neuropathic symptoms. We aimed to investigate the relationship between endothelial dysfunction and NO synthesis in type 2 diabetic patients with peripheral neuropathy after ALA treatment.Material and methodsFifty-four type 2 diabetic patients with neuropathy were included in the study. Serum ADMA concentration, ICAM-1, VCAM-1, oxidised low-density lipoprotein (oxLDL), and TNF-α levels were determined with Enzyme-Linked Immunosorbent Assay (ELISA). Nitric oxide concentration was measured by Griess reaction. Peripheral sensory nerve function was assessed by current perception threshold (CPT) testing. Autonomic function was assessed by Ewing’s five standard cardiovascular reflex tests composite autonomic score (CAS).ResultsAsymmetric dimethylarginine levels were significantly decreased (0.62 ±0.11 vs. 0.53 ±0.11 µmol/l, p &lt; 0.001), as well as TNF-α concentrations (1.21 ±0.42 pg/ml vs. 1.05 ±0.5 pg/ml, p &lt; 0.05), while NO levels were significantly increased (16.78 ±11.1 vs. 21.58 ±8.84 µmol/l, p &lt; 0.05) after six-months of 600 mg/day ALA treatment. VCAM-1, ICAM-1, and oxLDL levels did not change significantly. The CPT and CAS significantly improved after ALA treatment. The improvement of CPT values was correlated positively with the change of ADMA levels (r = 0.58, p &lt; 0.001). The change in ADMA level was more pronounced in responder patients based on both CPT and CAS.ConclusionsOur results suggest that ALA supplementation improves endothelial function characterised by serum levels of ADMA and TNF-α in patients with diabetic neuropathy. Changes in serum ADMA levels may predict the clinical response to ALA treatment.

Salt sensitivity and hypertension.

Salt sensitivity refers to the physiological trait present in mammals, including humans, by which the blood pressure (BP) of some members of the population exhibits changes parallel to changes in salt intake. It is commoner in elderly, females, Afro-Americans, patients with chronic kidney disease (CKD) and insulin resistance. Increased salt intake promotes an expansion of extracellular fluid volume and increases cardiac output. Salt-sensitive individuals present an abnormal kidney reaction to salt intake; the kidneys retain most of the salt due to an abnormal over-reactivity of sympathetic nervous system and a blunted suppression of renin-angiotensin axis. Moreover, instead of peripheral vascular resistance falling, salt-sensitive subjects present increased vascular resistance due mainly to impaired nitric oxide synthesis in endothelium. Recent studies have shown that part of the dietary salt loading accumulates in skin. Hypertensive and patients with CKD seem to have more sodium in skin comparing to healthy ones. However, we still have not fully explained the link between skin sodium, BP and salt sensitivity. Finally, although salt sensitivity plays a meaningful role in BP pathophysiology, it cannot be used by the physician in everyday patient's care, mainly due to lack of a simple and practical diagnostic test.