BACKGROUND AND PURPOSE Cystine‐knot miniproteins are characterized by a similar molecular structure. Some cystine‐knot miniproteins display therapeutically useful biological activities, as ...antithrombotic agents or tumour growth inhibitors. A critical event in the progression of tumours is the formation of new blood vessels. The aim of this work was to test two tomato cystine‐knot miniproteins for their effects on endothelial cell proliferation and angiogenesis in vitro.
EXPERIMENTAL APPROACH Two tomato cystine‐knot miniproteins (TCMPs) were expressed and purified either as recombinant or as native proteins from tomato fruits. The Matrigel assay was used to investigate the effects of TCMPs on in vitro angiogenesis. Viability and proliferation of endothelial cells were tested. Extracellular signal‐regulated kinase (ERK)1/2 phosphorylation was assayed in either HUVEC or A431 epidermal growth factor receptor (EGFR)‐overexpressing cells treated with TCMPs. EGFR phosphorylation was tested in A431 cells.
KEY RESULTS Both recombinant and native TCMPs inhibited in vitro angiogenesis of HUVEC cells at concentrations of 15–100 nM. The anti‐angiogenic effect of TCMPs was associated with the inhibition of ERK phosphorylation. The two miniproteins did not alter the viability and proliferation of the endothelial cells.
CONCLUSIONS AND IMPLICATIONS The anti‐angiogenetic properties of TCMPs are of potential pharmacological interest because they are common and natural components of the human diet, they possess low toxicity, they are active at submicromolar concentrations, they share a common molecular structure that can be used as a molecular platform for the design of molecules with enhanced biological activity.
OBJECTIVE:Plasma concentration of epoxyeicosatrienoic acids (EETs) derived from cytochrome P450 (CYP)-dependent metabolism of arachidonic acid is increased in women with preeclampsia (PE) as compared ...to normal pregnancy (N), and is even higher in fetal plasma (Herse et al. Circulation 2012, Jiang et al. Am J Hypertens 2013). We hypothesized that differences in EET synthesis or metabolism in the feto-placental unit underlie the observed differences in circulating EETs.
DESIGN AND METHOD:To evaluate EETs generation as well the expression of the relavant CYP isoforms and of the metabolizing enzyme soluble epoxide hydrolase (sEH), biopsies of placenta were collected from 19 N and 10 PE at the time of surgical delivery. EETs were extracted from tissue homogenates and analyzed by LCMS.
RESULTS:Both cis- and trans- EETs were detected in the placenta in PE and N, with similar mean ratios. Concentration of total EETs was higher in the placenta in PE compared to N (2.37 ± 1.42 ng/mg vs 1.20 ± 0.72 ng/mg, Mean ± SD, P < 0.01), especially the 5,6-, 8,9- and 11,12-EETs, measured in a subgroup of tissue samples (N = 10, PE = 5), were elevated. By immunohistochemistry, CYP2C8 was not detectable, CYP4A11 showed weak positivity in the mesenchimal axis of some villi (up to 50%) and scattered signal in the others. Also CYP2J2 was detectable in mesenchimal elements of placentas (scattered in 10–40% of villi, up to 50%). sEH showed weak signal in 1–3 cells for each villous, with a regular pattern distribution. CYP2C8, CYP4A11 and CYP2J2 were not detectable in umbilical cord. Western blotting analysis of placenta homogenates revealed a higher expression of sEH in N with respect to PE (3.9 ± 0.9 vs 0.8 ± 0.4 sEH relative expression, P < 0.05).
CONCLUSIONS:In conclusion, along with the enzymes implicated in their biosynthesis, significant amounts of EETs were found in the placenta and the umbilical cord. Reduced expression of sEH in PE may contribute to increased EET in the placenta. Altered synthesis of EETs occurs in the placenta, reinforcing the hypothesis of their pathogenetic role in PE.
In adults, low levels of vitamin D are associated with hypertension. The aim of this study was to evaluate the relationship between 24-h blood pressure (BP) patterns and vitamin D levels in obese ...children. We recorded anthropometric parameters, took blood samples for 25-hydroxivitamin D measurements and monitored ambulatory BP (ABP) in 32 obese children (male/female: 21/11, age 7-16 years). Subjects in the lower tertiles had higher homeostasis model assessment of insulin resistance, nighttime systolic and diastolic ABP, nighttime systolic and diastolic ABP load, 24-h ABP index and nighttime systolic and diastolic ABP index than those in the higher tertile. Vitamin D correlated negatively with 24-h and nighttime systolic ABP, 24-h systolic ABP load, nighttime systolic and diastolic ABP load, 24-h systolic ABP index and nighttime systolic ABP index. The percentage of subjects with pathological 24-h systolic BP (SBP) load, nighttime SBP load, nighttime diastolic BP (DBP) load, nighttime SBP index and nighttime DBP index increased progressively as the vitamin deficiency categories increased (χ(2)=10.26, P<0.05; χ(2)=16.34, P<0.01; χ(2)=10.23, P<0.05; χ(2)=10.38 and χ(2)=10.06, P <0.01). Low levels of vitamin D in obese children were associated with a higher BP burden, especially at night.
Background: Thromboembolism is a relatively common complication of chronic heart failure (HF) and the place of antiplatelet therapy is uncertain. Objectives: We characterized the rate of thromboxane ...and prostacyclin biosynthesis in chronic HF of ischemic origin, with the aim of separating the influence of HF on platelet activation from that of the underlying ischemic heart disease (IHD). Patients and Methods: We compared urinary 11‐dehydro‐thromboxane (TX)B2, 2,3 dinor 6‐keto‐PGF1α, 8‐iso‐prostaglandin (PG)F2α, and plasma N‐terminal pro‐brain natriuretic peptide (NT‐pro‐BNP), asymmetric dimethylarginine (ADMA), and soluble CD40 ligand (sCD40L), in 84 patients with HF secondary to IHD, 61 patients with IHD without HF and 42 healthy subjects. Results: HF patients not on aspirin had significantly higher urinary 11‐dehydro‐TXB2 as compared with healthy subjects (P < 0.0001) and IHD patients not on aspirin (P = 0.028). They also showed significantly higher 8‐iso‐PGF2α (P = 0.018), NT‐pro‐BNP (P = 0.021) and ADMA (P < 0.0001) than IHD patients not on aspirin. HF patients on low‐dose aspirin had significantly lower 11‐dehydro‐TXB2 (P < 0.0001), sCD40L (P = 0.007) and 2,3‐dinor‐6‐keto‐PGF1α (P = 0.005) than HF patients not treated with aspirin. HF patients in NYHA classes III and IV had significantly higher urinary 11‐dehydro‐TXB2 than patients in classes I and II, independently of aspirin treatment (P < 0.05). On multiple linear regression analysis, higher NT‐pro‐BNP levels, lack of aspirin therapy and sCD40L, predicted 11‐dehydro‐TXB2 excretion rate in HF patients (R2 = 0.771). Conclusions: Persistent platelet activation characterizes HF patients. This phenomenon is related to disease severity and is largely suppressable by low‐dose aspirin. The homeostatic increase in prostacyclin biosynthesis is impaired, possibly contributing to enhanced thrombotic risk in this setting.
Objective. Oxidative stress may be one of the important complex pathogenetic mechanisms that lead to damage in scleroderma; free radicals may provoke endothelial injury, fibroblast proliferation and ...fragmentation of autoantigens favouring induction of autoantibodies. The present study investigates the oxidant status in scleroderma patients by measuring the urinary concentration of 8-isoprostaglandin-F2α, an F2-isoprostane, and a product of free radical-mediated peroxidation of arachidonic acid. Methods. Forty-three scleroderma patients (42 women and 1 man, mean age 54.1 yr, mean disease duration 9.0 yr) underwent clinical evaluation and instrumental investigations in order to assess skin, vascular, lung and heart involvement. Von Willebrand factor was evaluated as marker of vascular dysfunction in 36 out of the 43 cases. The urinary level of 8-isoprostaglandin-F2α was measured in all scleroderma patients and in the 43 age- and sex-matched healthy controls. Results. Urinary levels of 8-isoprostaglandin-F2α were higher in scleroderma patients than in the healthy control group (341.7 vs 147.6 pg/mg creatinine; P<0.001). Values of 8-isoprostaglandin-F2α were strongly correlated with the nailfold videocapillaroscopy pattern and lung involvement (P=0.002 and 0.003, respectively), showing increasing levels with the progression of pulmonary severity. Correlation between 8-isoprostaglandin-F2α level and von Willebrand factor narrowly failed to reach statistical significance (P=0.05). There was no correlation between 8-isoprostaglandin-F2α concentration and disease activity, vascular, skin and heart involvement, disease pattern or autoantibody profile. Conclusions. Our study further supports the role of oxidant stress in the pathogenesis of scleroderma, showing a strong correlation between a marker of free radical damage with both the severity of lung involvement and the videocapillaroscopic patterns.
Background To determine if the common Pro12Ala polymorphism (rs1801282) of the peroxisome proliferator‐activated receptor (PPARG) gene is associated with the metabolic syndrome (MetS) or with its ...individual components in middle‐aged Swedish individuals.
Methods MetS was defined according to the National Cholesterol Education Program/Adult Panel III (NCEP/ATP III), the International Diabetes Federation (IDF) and the European Group for the Study of Insulin Resistance (EGIR) criteria in a population‐based sample of nearly 5000 subjects participating in the Malmö Diet and Cancer‐cardiovascular arm.
Results Of the subjects included in the analysis, 21.8, 29.4 and 20.4% had MetS according to the NCEP/ATP III, IDF and EGIR (only in subjects without diabetes) definitions, respectively. The Pro12Ala was not associated with MetS or with its individual components. These results were similar when patients with diabetes were excluded. Hypertensive and obese ala‐carriers had lower fasting glucose and hypertensive ala‐carriers also had lower level triglycerides (P < 0.05).
Conclusions Our data do not support a major role for the Pro12Ala variant of the PPARG gene in MetS and its individual components. The modest difference in triglyceride and glucose levels, restricted to hypertensive and obese subjects in our cohort, suggests that the polymorphism has a minor effect on glucose and lipid metabolism, particularly in individuals at risk for gluco‐metabolic disturbances.
Current literature suggests a higher risk of pregnancy-related complications in patients with renal fibromuscular dysplasia (FMD). The aim of our study was to assess the nature and prevalence of ...pregnancy-related complications in patients subsequently diagnosed with FMD. A call for participation was sent to centers contributing to the European/International FMD Registry. Patients with at least 1 pregnancy were included. Data on pregnancy were collected through medical files and FMD characteristics through the European/International FMD Registry. Data from 534 pregnancies were obtained in 237 patients. Despite the fact that, in 96% of cases, FMD was not diagnosed before pregnancy, 40% of women (n=93) experienced pregnancy-related complications, mostly gestational hypertension (25%) and preterm birth (20%), while preeclampsia was reported in only 7.5%. Only 1 patient experienced arterial dissection and another patient an aneurysm rupture. When compared with patients without pregnancy-related complications, patients with complicated pregnancies were younger at FMD diagnosis (43 versus 51 years old; P<0.001) and had a lower prevalence of cerebrovascular FMD (30% versus 52%; P=0.003) but underwent more often renal revascularization (63% versus 40%, P<0.001). In conclusion, the prevalence of pregnancy-related complications such as gestational hypertension and preterm birth was high in patients with FMD, probably related to the severity of renal FMD. However, the prevalence of preeclampsia and arterial complications was low/moderate. These findings emphasize the need to screen hypertensive women for FMD to ensure revascularization before pregnancy if indicated and appropriate follow-up during pregnancy, without discouraging patients with FMD from considering pregnancy.
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