Ramez Chahine

p-Hydroxybenzoate esters metabolism in MCF7 breast cancer cells

Parabens are among the most frequently used preservatives to inhibit microbial growth and extend the shelf life of a range of consumer products. The objective of the present study was to gain insight into the metabolism of parabens in breast cancer cells (MCF7) since they have demonstrated estrogenic activity towards these cells and have been detected in breast cancer tissues. The toxicity of parabens to MCF7 cells was determined using MTT assays. Hydrolysis of methyl-, butyl and benzyl-paraben to p-hydroxybenzoic acid was analyzed in cultured MCF7 cells and in cellular homogenates. Glucuronidation and sulfoconjugation were studied in MCF7 homogenates, and parabens were analyzed by HPLC. Methyl-paraben was shown to be far less toxic than butyl and benzyl-paraben. Parabens were completely stable in MCF7 homogenates whereas p-nitrophenyl acetate, a substrate type, underwent hydrolysis. MCF7 cell homogenates did not express glucuronidation and sulfoconjugation activities toward parabens. The higher stability of parabens may explain their accumulation in breast cancer tissue as previously reported in the literature.

Determination of antioxidant activity of saffron taken from the flower of Crocus sativus grown in Lebanon

Since oxidative stress has been implicated in most common cause of death, especially in case of cancer and cardiovascular disease, natural substances and spices that show antioxidant effects merit a closer examination. Saffron is the yellow natural spice derived from the flower of Crocus sativus and used as a coloring agent in many foods worldwide. In this study, we determined the total polyphenols content in the Lebanese saffron and the antioxidant effects of different extracts from this saffron in vitro using electrolysis of physiological solution for generation of free radicals (FR) in the presence of colorimetric indicator N,N-di-ethyl-P-phenylenedialanine; the absorbance was measured spectrophotometrically at 515 nm. Histophathological studies allowed us to observe the damages caused by FR in the isolated organs of hamsters (kidney, liver, lungs, and heart) and on the other hand the protection that saffron provided to these vital organs. By using assay kits, we evaluated the levels of lipid peroxidation and superoxide dismutase activity, the important free radical scavenging enzyme. The results showed that both boiled and soaked saffron at 0.45 mg/ml are highly effective against FR generated by electrolysis and against the damages caused to the organs tested as observed by light microscopy. Moreover, saffron significantly (p < 0.05) decreased lipid peroxidation and increased superoxide dismutase activity in all tissues used as compared to control. We concluded that Lebanese saffron strongly protects vital organs against oxidative stress.

Protective effect of saffron extract against doxorubicin cardiotoxicity in isolated rabbit heart

Abstract Context: Anticancer treatments such as anthracyclines are effective; however, they induce cardiotoxicity by releasing radical oxygen species (ROS). Saffron (Crocus sativus; Iridaceae) is a widely used spice with antioxidant properties and numerous health benefits that may provide cardioprotection. Objective: To assess the effect of saffron against acute myocardium damage by anthracyclines compared with electrolysis as a free radical generating system. Materials and methods: According to the Langendorff method, we used the model of an isolated rabbit heart perfused in retrograde. In one set of experiments, ROS was generated by electrolysis of the perfused heart solution (3 mA for 30 min) in the presence and absence of saffron extracts at the optimal dose (10 μg/ml). In another set, we perfused the heart with anthracycline, i.e. 30 μM doxorubicin (Doxo) in the presence and absence of 10 μg/ml saffron extracts. We evaluated cardiodynamics, as well as biochemical and pathological parameters, to emphasize the effectiveness of the treatment with saffron extract using the optimal dose of catalase (150 IU) as a positive control. Results: ROS generated, respectively, by electrolysis and by Doxo significantly (p < 0.05) affects cardiovascular function; it decreased ventricular pressure (45.02 and 40.41%), heart rate (36.31 and 22.39%) and coronary flow (50.98 and 36.67%). Increased lipid peroxidation of the myocardium was also observed (118.22 and 56.58%), while superoxide dismutase activity decreased (48.33 and 38.70%). The myocardial architecture was altered and the intercellular spaces increased. Conclusion: Saffron perfused during electrolysis helps trap ROS and significantly improves myocardial function; however, saffron was less effective against Doxo, thus suggesting that mechanisms other than oxidative stress underlie Doxo cardiotoxicity.

Saffron extracts alleviate cardiomyocytes injury induced by doxorubicin and ischemia-reperfusion in vitro

Abstract Doxorubicin (DOX), a highly active chemotherapeutic drug, faces limitations in clinical application due to severe cardiotoxic effects (mainly through increased oxidative stress). Therefore, its effect is exacerbated in subjects with ischemic heart disease. We have recently reported that saffron extract (SAF), a natural compound mainly consisting of safranal and corcins, exerts a protective effect against DOX oxidative cytotoxicity in isolated rabbit hearts. Here, we aimed to investigate whether SAF exerts cardioprotection against combined ischemia-reperfusion (I/R) and DOX toxicity in H9c2 cardiomyocytes. H9c2 were subjected to simulated I/R, with or without DOX treatment at reperfusion, in the presence or absence of SAF prior to ischemia or at reperfusion. We evaluated the effects of these treatments by MTT, LDH and western blot analysis. Apoptosis was assessed by Hoechst 33258 staining, tetramethyl rhodamine methyl ester fluorescence and caspase activity. The results showed that I/R and DOX significantly decreased cardiomyocytes viability, inhibited reperfusion injury salvage kinase cardioprotective pathway, reduced contractile proteins (α-Actinine, Troponine C and MLC), increased caspase-3 expression and induced loss of mitochondrial membrane potential. These effects were remarkably inhibited by treatment with SAF (10 μg/mL) at reperfusion. SAF activated AKT/P70S6K and ERK1/2, restored contractile proteins expression, inhibited mitochondrial permeability transition pore and decreased caspase-3 activity. In conclusion, our findings indicate that SAF treatment exerted cardioprotection against I/R and DOX toxicity by reducing oxidative stress (LDH assay). Thereby, SAF offers a potential novel antioxidant therapeutic strategy to counteract I/R and DOX cardiotoxicity, paving the way for future clinical trials.

Prevalence of stroke symptoms among stroke‐free residents: first national data from Lebanon

Background Stroke symptoms are common among people without a history of stroke or transient ischemic attack. Reported stroke symptoms may represent stroke episodes that failed to reach the threshold for clinical diagnosis. Aims This study aimed to assess in the Lebanese population the prevalence of self-reported stroke symptoms in a stroke-and transient ischemic attack-free population, and the association of these symptoms with major risk factors for stroke. Methods We carried out a cross-sectional study using a multistage cluster sample across Lebanon. We interviewed residents aged 40 years and more. Stroke symptoms were assessed using the Questionnaire for Verifying Stroke-Free Status. Results We included 1515 individuals (mean age was 57.2 ± 12.4 years, 783 women, 51.7%). Among 1460 participants stroke- and transient ischemic attack-free, 175 had experienced at least one stroke symptom (12.1%, 95% CI 9.9%-14.3%). Arterial hypertension (adjOR 4.37, 95% CI 2.68-7.12), history of heart disease (adjOR 3.34, 95% CI 2.00-5.56), current waterpipe smoking (adjOR 3.88, 95% CI 2.33-6.48), current and former cigarette smoking (adjOR 1.84, 95% CI 1.18-2.87 and adjOR 2.01, 95% CI 1.13-3.5, respectively), psychological distress (adjOR 1.04, 95% CI 1.02-1.05), the Mediterranean diet score (adjOR 0.87, 95% CI 0.76-0.99), and regular physical activity (adjOR 0.45, 95% CI 0.26-0.77) were independently associated with stroke symptoms. Conclusions This is the first study conducted in the Middle East, assessing self-reported stroke symptoms among stroke-free residents. Our study showed that almost one in eight residents without a history of stroke or transient ischemic attack has had stroke symptoms. Major vascular risk factors are associated with these symptoms, thus allowing for prevention strategies.

Cardioprotective effect of saffron extracts against acute doxorubicin toxicity in isolated rabbit hearts submitted to ischemia-reperfusion injury.

Abstract Doxorubicin (DOX) is an anthracycline antibiotic routinely used as a chemotherapeutic agent for the treatment of solid tumours. However, DOX possesses an acute and cumulative cardiotoxicity due to free radical production. The present study was designed to investigate the possible protective effects of saffron (Crocus sativus) extracts against DOX-induced acute cardiotoxicity in isolated rabbit hearts submitted to 30 min global ischemia followed by 40 min reperfusion. DOX was delivered during reperfusion, without or with saffron given 5 min before ischemia or at reperfusion. Cardiodynamic, biochemical, and histopathological parameters were determined. In addition, to determine the expression of the AKT/mTOR/4EBP1 pathway, the levels of p38 MAPK and cardiac troponin T in heart homogenates were visualized by Western blotting. DOX administration during 40 min of reperfusion increased ischemic tissue damage, but did not act synergistically. Administration of saffron extracts during the first minutes of reperfusion significantly reduced oxidative myocardial damage, but was less effective when given before ischemia. Subsequent Western blot analysis revealed that saffron administration preserved cardiac troponin T proteins, inhibited the p38 MAPK pathway, and activated the AKT/mTOR/4EBP1 pathway in reperfusion- and DOX-treated rabbit hearts. In conclusion, saffron extracts, acting through antioxidant and antiapoptotic mechanisms, exhibited a protective effect against DOX-induced cardiotoxicity under ischemic condition.

Predictors of Uncontrolled Blood Pressure in Treated Hypertensive Individuals: First Population‐Based Study in Lebanon

Arterial hypertension is a leading risk factor for cardiovascular disease and stroke. This study aimed to assess the predictors of uncontrolled systolic and diastolic blood pressure (BP) in Lebanon among treated hypertensive individuals. The authors included 562 participants 40 years and older. The potential predictors included sociodemographic characteristics, self‐reported health information, and medication adherence. Prevalence of uncontrolled systolic and diastolic BP reached 43.1% and 24.9%, respectively. Independent predictors of uncontrolled systolic BP were older age, male sex, and low and medium medication adherence level. Predictors of uncontrolled diastolic BP were younger age, obesity, and low medication adherence level. Married individuals and patients taking statins had better diastolic BP control. Uncontrolled BP is a major public health problem in Lebanon. The authors identified low adherence as a major modifiable risk factor for systolic and diastolic BP control and obesity as a major modifiable risk factor for diastolic BP control.

Nicotine metabolism in healthy smokers and patients with cardiovascular diseases

In this study, we measured the excretion rate of nicotine and its two major metabolites, cotinine and trans-3′-hydroxycotinine (THOC), in the urine of 25 healthy smokers and 15 smokers who underwent a coronary artery bypass surgery or coronary angioplasty. After 1 day of smoking cessation, urine samples were collected in the morning, before smoking two cigarettes, and then three times after smoking, approximately 4 h apart. The results show that (i) in healthy smokers, nicotine and its two major metabolites were present at high concentration in the first urine sample after smoking, (ii) in smokers with cardiovascular disease nicotine and cotinine were less excreted whereas THOC was more excreted, mainly in the second urine sample. We conclude that this shift in nicotine metabolism may contribute to smoking-induced cardiovascular disease. (Mol Cell Biochem xxx: 241–244, 2005)

Gamma-irradiated ceruloplasmin affords antifibrillatory protection against ischemia/reperfusion damage in the isolated rat heart

Purpose: Ceruloplasmin (CP), an important serum antioxidant, was previously found to reduce the incidence of ventricular fibrillation (VF) induced by ischemia and reperfusion in isolated rat hearts. The present study investigated whether CP sterilized by gamma-irradiation maintains its antiarrhythmic capacity and in vitro antioxidant properties. Materials and methods: Isolated rat hearts submitted to regional ischemia (15 min), were reperfused (10 min) with native CP or with CP irradiated at various doses (1–3 kGy) in the absence or presence of tyrosine (Tyr). Results: All untreated hearts showed VF at reperfusion, which were all irreversible ventricular fibrillation (IVF). No IVF were found in hearts treated with native CP or gamma-irradiated CP. Cardioprotection afforded by irradiated CP (with or without Tyr) was slightly higher than that obtained with native CP. No VF at all (100% prevention) was found in hearts treated with CP irradiated alone or in the presence of tyrosine at 3 kGy. Tyrosine and irradiated tyrosine had no cardiotoxic or protective effects on reperfusion-induced arrhythmias. The Oxygen Radical Absorbing Capacity (ORAC), measured in vitro with β-phycoerythrin (β-PE) fluorescent indicator, was slightly higher for gamma-irradiated CP in the presence of Tyr. Conclusions: Ceruloplasmin sterilized by gamma-irradiation maintains antioxidant and antiarrhythmic effects in the post-ischemia reperfused isolated rat heart.

Association between vitamin D deficiency and lipid and non-lipid markers of cardiovascular diseases in the middle east region

BackgroundPrevious studies have associated vitamin D deficiency with cardiovascular disease (CVD) markers. The underlying mechanism remains elusive. Lipid and non-lipid markers of CVD and their relationship to vitamin D deficiency have not been assessed simultaneously.ObjectiveTo measure the association between vitamin D deficiency and non-lipid markers of CVD after adjustment of lipid markers.MethodsThis cross-sectional study used the following biological data, which was routinely collected in a general hospital laboratory database between 2011 and 2016: 25OH vitamin D [25(OH)D], creatinine, CKD-EPI eGFR (eGFR), fasting blood glucose (FPG), glycated hemoglobin (HbA1c), uric acid, γ-glutamyl transferase (γGT), C-reactive protein (CRP), total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, and a surrogate for CVD. Crude odds ratios (ORs) and ORs adjusted for lipid profile, gender and age using separate logistic regression models were derived.ResultsA total of 8658 subjects were included. Half had 25(OH)D < 20 ng/mL. 25(OH)D was associated with increased odds of CRP, eGFR, increased uric acid, γGT, FPG, HbA1c, male gender, CV status, and abnormal lipid markers. After adjustment for lipid markers, age, and gender, vitamin D deficiency was associated with increased odds of CRP, eGFR, γGT, FPG, HbA1c, and the surrogate for CVD.ConclusionsIn this exploratory analysis, the first of its kind in the MENA region, vitamin D deficiency was associated with abnormal lipid markers, non-lipid markers of CVD, male gender, lower eGFR, and a surrogate variable for CVD. The association between vitamin D deficiency and non-lipid markers of CVD persisted after adjustment for lipid markers, age, and gender.