Vincenzo Arcoraci

17β-oestradiol reduces cardiac leukocyte accumulation in myocardial ischaemia reperfusion injury in rat

Abstract We investigated whether oestrogens modulate the phenomenon of leukocyte accumulation during ischaemia and reperfusion of the myocardium. Anaesthetized rats were subjected to total occlusion (1 h) of the left main coronary artery followed by 1 h reperfusion. Sham myocardial ischaemia–reperfusion rats (Sham) were used as controls. Myocardial necrosis, myocardial myeloperoxidase activity, serum creatinine phosphokinase activity, serum and macrophages tumour necrosis factor (TNF-α) and the myocardial staining of intercellular adhesion molecule-1 (ICAM-1) were evaluated. Myocardial ischaemia plus reperfusion in untreated rats produced marked myocardial necrosis, increased serum creatinine phosphokinase activity (348±38 U/ml) and cardiac myeloperoxidase activity, a marker of polymorphonuclear leukocyte accumulation, both in the area at risk and in the necrotic area (MPO; 9±1.1 U×10−3/g tissue and 8.2±1 U×10−3/g tissue, respectively), and induced a marked increase in the macrophage (156±14 U/ml at the end of reperfusion) and serum (344±12 U/ml, at the end of reperfusion) levels of TNF-α. Finally, myocardial ischaemia–reperfusion injury increased ICAM-1 staining in the myocardium. Administration of 17β-oestradiol (5, 10 and 20 μg/kg, i.m. 5 min after induction of myocardial ischaemia–reperfusion injury), lowered myocardial necrosis and myeloperoxidase activity in the area at risk and in the necrotic area, reduced serum and macrophages TNF-α (20±3 U/ml and 9±3 U/ml, respectively) and decreased serum creatinine phosphokinase activity (67±3 U/ml). Oestrogen treatment also blunted the increased staining of ICAM-1 in the injured myocardium. Finally, 17β-oestradiol added in vitro to peritoneal macrophages collected from untreated rats subjected to myocardial ischaemia–reperfusion injury, significantly reduced TNF-α production. Our results suggest that 17β-oestradiol, by inhibiting TNF-α production, limits the deleterious ICAM-1-mediated binding of leukocytes to injured mycardium and protects against myocardial ischaemia–reperfusion injury.

Food deprivation increases brain nitric oxide synthase and depresses brain serotonin levels in rats

We studied nitric oxide (NO) synthase activity and serotonin content in the diencephalon of 24 hr food deprived rats. NO synthase activity was significantly increased whereas serotonin levels together with those of tryptophan and 5-hydroxyindoleacetic acid (5-HIAA) were reduced in food deprived rats when compared to control rats. NG-Nitro-L-arginine (L-NO Arg), an inhibitor of NO synthase, was used as a tool to study the role of NO in food deprivation. Twenty-four hr food deprived male Sprague-Dawley rats were intraperitoneally (i.p.) administered L-NO Arg (12.5, 25 and 50 mg/kg) before food presentation. Control rats received a NaCl (0.9%) solution. Food consumption was monitored 1 and 2 hr after food presentation. L-NO Arg administration produced a dose-dependent reduction in food intake. Pretreatment with metergoline (2 mg/kg) but not with ritanserin (1 mg/kg) antagonized the anorectic effect of L-NO Arg. Moreover, in the diencephalon L-NO Arg significantly reduced NO synthase activity whereas it increased serotonin levels. Our data indicate that NO might have a physiological role in the regulation of food intake and suggest that brain NO may modulate the central serotoninergic system.

Central serotoninergic system involvement in the anorexia induced by NG-nitro-L-arginine, an inhibitor of nitric oxide synthase.

The effects of NG-nitro-L-arginine, an inhibitor of brain nitric oxide (NO) synthase, on central serotoninergic system were studied in male obese Zucker rats and in their lean age-matched controls (FA/?; FA/FA), both groups aged 14 weeks. Acute injection of NG-nitro-L-arginine (50 mg/kg i.p.) or repeated administration of NG-nitro-L-arginine (50 mg/kg i.p. daily, for 7 days) reduced food intake and body weight in obese rats. Acute administration of NG-nitro-L-arginine reduced food intake in lean rats. However, lean rats showed tolerance to the NG-nitro-L-arginine effects after repeated administration. NG-Nitro-L-arginine administration significantly increased serotonin metabolism in the cortex, diencephalon and medulla-pons of obese Zucker rats after either acute or repeated administration of NG-nitro-L-arginine. In contrast, NG-nitro-L-arginine increased serotonin metabolism in lean rats only after acute administration, and the appearance of tolerance to NG-nitro-L-arginine anorectic effects paralleled the failure of NG-nitro-L-arginine to increase serotonin metabolism. The present data extend our previous findings indicating that NG-nitro-L-arginine possesses anorectic activity in obese Zucker rats, and clearly suggest that the central serotoninergic system mediates the anorexia induced by inhibitors of brain NO synthase.

Genistein in the Metabolic Syndrome: Results of a Randomized Clinical Trial

CONTEXT This study was performed to evaluate the effects of genistein on metabolic and cardiovascular risk factors in Caucasian postmenopausal subjects with metabolic syndrome (MetS). OBJECTIVE Our objective was to assess the effects of genistein on surrogate endpoints associated with diabetes and cardiovascular disease. DESIGN AND SETTING This was a randomized, double-blind, placebo-controlled trial at 3 university medical centers in Italy. PATIENTS Patients included 120 postmenopausal women with MetS according to modified Third Report of the National Cholesterol Education Program (NCEP), Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) criteria. INTERVENTION After a 4-week stabilization period, postmenopausal women with MetS were randomly assigned to receive placebo (n = 60) or 54 mg genistein daily (n = 60) for 1 year. MAIN OUTCOME MEASURES The primary outcome was homeostasis model assessment for insulin resistance (HOMA-IR) at 1 year. Secondary outcomes were fasting glucose, fasting insulin, total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides, visfatin, adiponectin, and homocysteine levels. Data on adverse events were also recorded. RESULTS At 1 year in genistein recipients, fasting glucose, fasting insulin, and HOMA-IR (mean from 4.5 to 2.7; P < .001) decreased and were unchanged in placebo recipients. Genistein statistically increased HDL-C (mean from 46.4 to 56.8 mg/dL) and adiponectin and decreased total cholesterol, LDL-C (mean from 108.8 to 78.7 mg/dL), triglycerides, visfatin, and homocysteine (mean from 14.3 to 11.7 μmol/L) blood levels. Systolic and diastolic blood pressure was also reduced in genistein recipients. Genistein recipients neither experienced more side adverse effects than placebo nor discontinued the study. CONCLUSION One year of treatment with genistein improves surrogate endpoints associated with risk for diabetes and cardiovascular disease in postmenopausal women with MetS.

The Effect of PDRN, an Adenosine Receptor A2A Agonist, on the Healing of Chronic Diabetic Foot Ulcers: Results of a Clinical Trial

CONTEXT Foot ulcer is the principal cause of hospitalization for patients with diabetes. Polydeoxyribonucleotide (PDRN), an adenosine A2A receptor agonist, improves wound healing in diabetic mice. OBJECTIVE The aim of this study was to evaluate the effect of PDRN on chronic ulcer healing in patients with diabetes. DESIGN AND SETTING This randomized, double-blind, placebo-controlled trial, involved two medical centers in Italy. INTERVENTION Patients with diabetes showing hard-to-heal ulcers (Wagner grade 1 or 2) were randomly assigned to receive placebo (n = 106) or PDRN (n = 110). The treatments (PDRN and placebo) were performed for 8 weeks by intramuscular and perilesional route [corrected]. MAIN OUTCOME MEASURES The primary outcome was complete ulcer healing. Secondary outcomes were the days needed to complete wound closure and the reepithelialization of wound surface (as percentage of the original area). RESULTS After 8 weeks, 91 placebo and 101 PDRN subjects completed the study. Complete healing was achieved in 18.9% [95% confidence interval (CI) 11.4-26.3] of placebo and in 37.3% (95% CI 28.2-46.3) of PDRN-treated patients (P = .0027). After 8 weeks, PDRN increased the closure of foot ulcers in diabetic subjects (hazard ratio 2.20; 95% CI 1.29-3.75; P = .004). The median time to complete wound healing was 49 days for placebo (range 28-56 d) and 30 days for PDRN-treated subjects (range 14-56 d; P = .0027). The median epithelialized area of the ulcers (expressed as percentage) was 49.3% in the placebo and 82.2% in the PDRN group (P < .001). CONCLUSIONS PDRN facilitates the healing of Wagner 1 or 2 diabetic foot ulcers.

Effects of aglycone genistein in a rat experimental model of postmenopausal metabolic syndrome

Genistein aglycone, a soy derived isoflavone, has been demonstrated to be effective in reducing cardiovascular risk in postmenopausal women. We therefore investigated its effects in an experimental model of postmenopausal metabolic syndrome. Female spontaneously hypertensive obese rats (SHROB, n=40), a genetic model of syndrome X, and age-matched Wistar Kyoto (WKY, n=40) rats were used. A group of SHROB (n=20) and WKY (n=20) animals were ovariectomized (OVX). Four weeks after surgery all animals were randomized to receive either genistein (54 mg/human equivalent dose/day for 4 weeks), or vehicle. Body weight, food intake, systolic blood pressure (SBP), heart rate, plasma glucose, insulin resistance (HOMA-IR), total plasma cholesterol and triglycerides, and uterine weights were studied. Furthermore, we investigated acetylcholine- and sodium nitroprusside-induced relaxation of aortic rings as well as NG-L-arginine (L-NMA: 10-100 mM) induced vasoconstriction in phenylephrine-precontracted aortic segments. Liver expression of the peroxisome proliferator-activated receptor alpha (PPARA and gamma (PPARG was also assessed. OVX animals had a slight increase in SBP, body weight, insulin resistance, and plasma cholesterol. OVX-SHROB rats showed also impaired endothelial responses, blunted L-NMA induced contraction (L-NMA 100 mM, WKY=2.2+/-0.3 g/mg tissue; OVX-SHROB=1.1+/-0.4 g/mg tissue). Genistein treatment decreased SBP and plasma lipids, ameliorated endothelial dysfunction and insulin resistance, increased HDL cholesterol, and enhanced liver expression of PPARA and PPARG. Our data suggest that genistein is effective in ameliorating cardiovascular profiles in an experimental model of postmenopausal metabolic syndrome, attenuating the features of this disease. The effects of genistein are likely mediated by PPARA and PPARG receptors. This evidence would support the rationale for some pilot clinical trials using genistein in postmenopausal women affected by metabolic syndrome.

ROS-Mediated NLRP3 Inflammasome Activation in Brain, Heart, Kidney, and Testis Ischemia/Reperfusion Injury

Ischemia and reperfusion (I/R) causes a reduction in arterial blood supply to tissues, followed by the restoration of perfusion and consequent reoxygenation. The reestablishment of blood flow triggers further damage to the ischemic tissue through reactive oxygen species (ROS) accumulation, interference with cellular ion homeostasis, and inflammatory responses to cell death. In normal conditions, ROS mediate important beneficial responses. When their production is prolonged or elevated, harmful events are observed with peculiar cellular changes. In particular, during I/R, ROS stimulate tissue inflammation and induce NLRP3 inflammasome activation. The mechanisms underlying the activation of NLRP3 are several and not completely elucidated. It was recently shown that NLRP3 might sense directly the presence of ROS produced by normal or malfunctioning mitochondria or indirectly by other activators of NLRP3. Aim of the present review is to describe the current knowledge on the role of NLRP3 in some organs (brain, heart, kidney, and testis) after I/R injury, with particular regard to the role played by ROS in its activation. Furthermore, as no specific therapy for the prevention or treatment of the high mortality and morbidity associated with I/R is available, the state of the art of the development of novel therapeutic approaches is illustrated.

Newer and older antiepileptic drug use in Southern Italy: A population-based study during the years 2003–2005

AIM To analyse the prescribing pattern of newer and older antiepileptic drugs (AEDs) during the years 2003-2005. METHODS From the Caserta-1 Local Health Service database, 93 general practitioners (GPs) were recruited. Among 127,389 individuals aged > or =15 years registered in the lists of these GPs, we selected patients who received at least one AED prescription during the study period. Use of newer and older AEDs was calculated as 1-year prevalence and incidence as well as defined daily dose (DDD) per 1000 inhabitants/day. Sub-analyses by gender, age and indication of use were performed. RESULTS Overall, prevalence and incidence of use remained stable for older AEDs, while it strongly increased for newer AEDs. In particular, 25% increase of incident treatments with newer AED have been reported from 2004 to 2005. The total volume of AED use remained stable during the study years, despite the proportion of newer AEDs slightly increased (from 24.6% in 2003 to 30.1% in 2005). The main indication of use was epileptic disorders for older AEDs (56% of users), and neuropathic pain for newer AEDs (69%). CONCLUSIONS Prevalence and incidence of use of newer AED strongly increased during the years 2003-2005 in a general practice of Southern Italy. Significant differences are shown in the prescribing pattern of newer and older medications: older AEDs are mainly used in the treatment of epileptic disorders, while newer compounds are preferred for conditions other than epilepsy, in particular neuropathic pain.

Prescribing pattern of drugs in the treatment of osteoarthritis in italian general practice: The effect of rofecoxib withdrawal

OBJECTIVE In October 2004, rofecoxib was removed from the world market because of an increased risk of myocardial infarction. The aim of the present study was to compare the trend of nonsteroidal antiinflammatory drug (NSAID) use and other analgesics in osteoarthritis (OA) treatment before and after rofecoxib withdrawal in Italian general practice. METHODS From the Caserta-1 Local Health Service database, 97 general practitioners were recruited. Prevalence and incidence of use of any study drug were calculated within 1 year before and after rofecoxib withdrawal. RESULTS One-year prevalence of nonselective and preferential NSAID use did not change after rofecoxib withdrawal, whereas coxib use fell from 4.4% (95% confidence interval [95% CI] 4.2-4.5%) in the period before rofecoxib withdrawal (period I) to 1.6% (95% CI 1.5-1.7%) in the period after withdrawal (period II). Weak opioids were used in no more than 0.4% (95% CI 0.3-0.5%) in period II, after their introduction to reimbursement in December 2004. Also, 1-year incidence of coxib decreased from 31.3 per 1,000 (95% CI 30.2-32.4%) in period I to 8.7 per 1,000 (95% CI 8.1-9.2%) in period II. The disappearance of rofecoxib was associated with replacement drugs such as newly marketed dexibuprofen and aceclofenac, whereas nimesulide use coincidentally decreased. CONCLUSION Rofecoxib withdrawal has markedly changed the prescribing pattern of drugs that are used in OA-related pain treatment, with a striking decrease of coxib use in Italian general practice. Education strategies addressed to health professionals should be planned to improve the management of pain treatment, particularly in degenerative joint diseases.

Genistein and endothelial function in postmenopausal women with metabolic syndrome.

Previous data have suggested that genistein could exert beneficial effects on endothelial function and on predictors of cardiovascular risk in healthy postmenopausal women. In a randomized clinical trial, we studied the effects of genistein on endothelial function in postmenopausal women with metabolic syndrome (MS).