A novel synthetised sulphonylhydrazone coumarin (E)-4-methyl-N'-(1-(3-oxo-3H-benzo[f]chromen-2- yl)ethylidene)benzenesulphonohydrazide protect against isoproterenol-induced myocardial infarction in rats by attenuating oxidative damage, biological changes and electrocardiogram.

Coumarins and their derivatives are becoming a potential source for new drug discovery due to their vast array of biological activities. The present study was designed to investigate the cardioprotective effects of a newly synthesised coumarin, symbolised as 5,6-PhSHC, against cardiac remodelling process in isoproterenol (ISO) induced myocardial infarction (MI) in male Wistar rats by evaluating haematological, biochemical and cardiac biomarkers. Rats were pre/co-treated with 5,6-PhSHC or clopidogrel (150 µg/kg body weight) daily for a period of 7 days and then MI was induced by injecting ISO (85 mg/kg body weight), at an interval of 24 hours for 2 consecutive days, on the sixth and seventh days. The in vivo exploration indicated that the injection of 5,6-PhSHC improved the electrocardiographic (ECG) pattern and prevented severe heart damage by reducing leakage of the cardiac injury markers, such as troponin-T (cTn-T), lactate dehydrogenase (LDH), and creatine kinase-MB. The cellular architecture of cardiac sections, altered in the myocardium of infracted rats, was reversed by 5,6-PhSHC treatment. Results showed that injection of 5,6-PhSHC elicited significant cardioprotective effects by prevention of myocardium cell necrosis and inflammatory cells infiltration, along with marked decrease in plasma levels of fibrinogen. In addition, the total cholesterol, triglyceride, LDL-c, and HDL profiles underwent remarkable beneficial changes. It was also interesting to note that 5,6-PhSHC enhanced the antioxidative defence mechanisms by increasing myocardial glutathione (GSH) level, superoxide dismutase (SOD), and catalase (CAT) activities, together with reducing the levels of thiobarbituric-acid-reactive substances (TBARS), when compared with ISO-induced rats. Taken together, these findings suggested a beneficial role for 5,6-PhSHC against ISO-induced MI in rats. Furthermore, in silico analysis showed that 5,6-PhSHC possess high computational affinities (E-value >-9.0 kcal/mol) against cyclooxygenase-2 (PDB-ID: 1CX2), vitamin K epoxide reductase (PDB-ID: 3KP9), glycoprotein-IIb/IIIa (PDB-ID: 2VDM) and catalase (PDB-ID: 1DGF). Therefore, the present study provided promising data that the newly synthesised coumarin can be useful in the design and synthesis of novel drug against myocardial infarction.

Oleuropein Protects Against Cerebral Ischemia Injury in Rats: Molecular Docking, Biochemical and Histological Findings.

This study was designed to evaluate the underlying protective mechanisms of oleuropein involved in alleviating brain damage in a rat model of ischemic stroke. Male Wistar rats were divided into four groups; Control, stroke (MCAO), MCAO + clopidogrel (Clop) and MCAO + oleuropein (Ole). Results showed that the MCAO group evidenced significant brain edema (+ 9%) as well as increases of plasma cardiac markers such as lactate deshydrogenase (LDH), creatine kinase (CK-MB), fibrinogen and Trop-T by 11 %, 43%, 168 and 590%, respectively, as compared to the control group. Moreover, infarcted rats exhibited remarkable elevated levels of angiotensin converting enzyme (ACE), both in plasma and brain tissue, with astrocyte swelling and necrotic neurons in the infarct zone, hyponatremia, and increased rate of thiobarbituric acid-reactive substances (TBARS) by 89% associated with decreases in the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (Cat) by 51%, 44 and 42%, respectively, compared to normal control rats. However, MCAO rats treated with oleuropein underwent mitigation of cerebral edema, correction of hyponatremia, remarkable decrease of plasma fibrinogen and cardiac dysfunctional enzymes, inhibition of ACE activity and improvement of oxidative stress status in brain tissue. Furthermore, in silico analysis showed considerable inhibitions of ACE, protein disulfide isomerase (PDI) and TGF-ß1, an indicative of potent anti-embolic properties. Overall, oleuropein offers a neuroprotective effect against ischemic stroke through its antioxidative and antithrombotic activities.

Cardiopreventive capacity of a novel (E)-N'-(1-(7-methoxy-2-oxo-2H-chromen-3-yl) ethylidene)-4-methylbenzenesulfonohydrazide against isoproterenol-induced myocardial infarction by moderating biochemical, oxidative stress, and histological parameters.

This study is carried out to assess the cardiopreventive effect of (E)-N'-(1-(7-methoxy-2-oxo-2H-chromen-3-yl) ethylidene)-4-methylbenzenesulfonohydrazide or SHC, a novel synthesized coumarin, against myocardial infarction induced by isoproterenol (ISO). The SHC compound was identified and characterized by spectral methods (infrared, 1 H NMR [nuclear magnetic resonance], 13 C NMR, Nuclear Overhauser Effect Spectroscopy, and high-resolution mass spectroscopy). Male Wistar rats were divided into four groups: Control, ISO (rats were injected subcutaneously by 85 mg/kg body weight [BW] of isoproterenol at Days 6 and 7 of the experience), ISO + SHC (150 µg/kg BW, orally for 7 days) and ISO + acenocoumarol (150 µg/kg BW, orally for 7 days). Results showed that ISO induced a remarkable alteration of electrocardiogram (ECG) pattern and increases of plasma cardiac troponin T, creatine kinase-MB, total cholesterol, triglycerides, low-density lipoprotein-cholesterol, lactate dehydrogenase, aspartate transaminase, and malondialdehyde. In addition, ISO reduced the high-density lipoprotein-cholesterol content and the activities of superoxide dismutase and glutathione peroxidase, with the induction of myocardial necrosis. However, SHC administration revealed a significant decrease in cardiac dysfunction markers, restored normal ECG pattern, as well as improving lipids parameters. Moreover, SHC treatment remarkably alleviated the cardiac oxidative stress and the myocardial remodeling process. Overall, the SHC offers good protection from acute myocardial infarction through the antioxidant capacity.

Anti-embolic and anti-oxidative effects of a novel (E)-4-amino-N'-(1-(7-hydroxy-2-oxo-2H-chromen-3-yl) ethylidene) benzohydrazide against isoproterenol and vitamin-K induced ischemic stroke.

This study aimed to evaluate the cerebroprotective potential of a novel synthetic coumarin, (E)-4-amino-N'-(1-(7-hydroxy-2-oxo-2H-chromen-3-yl)ethylidene) benzohydrazide noted (HC) against a pharmaceutically induced ischemic stroke in experimental male Wistar rats. Animals were randomly allocated into four groups: control, Stroke, Stroke + Ace (acenocoumarol) and Stroke + HC-treated group for 7 days. Our results showed that stroke group evidenced atrial flutter, significant cardiac hypertrophy (+23%) and increase in plasma level of troponin-T, with disturbance in plasma ionic levels and rise in fibrinogen rate and oxidative damages in heart and brain. Moreover, the histological findings revealed myocardium necrosis, cardiac cavity thrombi and brain injury as compared to normal rats. However, HC-treatment significantly prevents the embolic process, improves cerebral damages and mitigates the oxidative stress markers in stroke rats. Overall, HC is endowed with a thrombolytic potential against MI and stroke in such severe conditions through an anti-vit K (AVK) mechanism.

Potential Hepatoprotective Activity of Super Critical Carbon Dioxide Olive Leaf Extracts against CCl4-Induced Liver Damage.

Virgin olive oil has demonstrated its effective activity against oxidative stress. However, data on the bioactive effect of olive leaves or their major constituents on the liver are scarce. The present research work was conducted to evaluate the hepatoprotective effects of supercritical carbon dioxide (SC-CO2) extracts from fresh and dried olive leaves on hepatotoxicity caused by carbon tetrachloride (CCl4) in rat models. For this purpose, healthy albino rats of 180-250 g weight were used. The assessment of biochemical markers was carried out on blood and liver tissue. Then, a histopathological study was carried out on liver tissue. The obtained results showed that fresh and dried olive leaf extracts ameliorate the perturbed biochemical parameters caused by CCl4 treatment. Furthermore, the results registered for the histopathological study are in accordance with the biochemical parameters and the protective capacity of SC-CO2 extracts against DNA damage, indicating that olive leaf extracts helped to improve liver fibrosis caused by CCl4 treatment.

(E)-N'-(1-(3-oxo-3H-benzo[f]chromen-2-yl)ethylidene)benzohydrazide protecting rat heart tissues from isoproterenol toxicity: Evidence from in vitro and in vivo tests.

The current study was aimed to assess the protective effect of a new molecule (E)-N'-(1-(3-oxo-3H-benzo[f]chromen-2-yl)ethylidene)benzohydrazide, denoted 1c, against cardiac remodeling process in isoproterenol (Isop) induced myocardial infarction (MI) in rats. Male Wistar rats were randomly divided into four groups, control, Isop (85 mg/kg body weight was injected subcutaneously into rats at an interval of 24 h for 2 days (6th and 7th day) to induce MI and pretreated animals with acenocoumarol (Ace) (150 µg/kg bw) and 1c (150 µg/kg bw) by oral administration during 7 days and injected with isoproterenol (Isop + Ace) and (Isop + 1c) groups. Results in vitro showed that 1c is endowed with potent inhibition of angiotensin-converting enzyme (ACE) with an IC50 39.12 µg/ml. The in vivo exploration evidenced alteration in the ECG pattern, notable cardiac hypertrophy and increase in plasma level of fibrinogen, troponin-T, CK-MB and LDH, AST and ALT by 171%, 300%, 50%, 64% and 75% respectively with histological myocardium necrosis and cells inflammatory infiltration. However, pre-treatment with 1c improved the ECG pattern reduced significantly the cardiac dysfunction markers and ameliorated the thrombolytic process by decreasing fibrinogen level as compared to untreated infracted rats. Overall, (E)-N'-(1-(3-oxo-3H-benzo[f]chromen-2-yl)ethylidene)benzohydrazide 1c could be used as anticoagulant agent to prevent thrombosis in acute myocardial infarction.

Zygophyllum album leaves extract prevented hepatic fibrosis in rats, by reducing liver injury and suppressing oxidative stress, inflammation, apoptosis and the TGF-ß1/Smads signaling pathways. Exploring of bioactive compounds using HPLC-DAD-ESI-QTOF-MS/MS.

Zygophyllum album is traditionally used against many illnesses, such as liver disease. The present study investigated the bioactive compounds in methanol extract of Z. album (MEZA) using HPLC-DAD-ESI-QTOF-MS/MS and explored its possible antioxidative, anti-inflammatory, anti-apoptotic, and hepatoprotective effect. Twelve phenolic compounds were identified; isorhamnetin-3-O-rutinoside being the main one was the main composite (144.6 mg/100 g dm). Results showed that MEZA reduced significantly the biochemical markers (AST, ALT, LDH and ALP), and the hepatic oxidative stress indicators (MDA, PC, SOD, CAT, and GPx) in deltamethrin (DLM)-treated rats. Moreover, MEZA limited the inflammatory responses through downregulation of NF-κB gene, which suppressed the production of proinflammatory cytokines (TNF-α, IL-1ß, IL-6). Furthermore, Z. album reduced DLM-induced apoptosis by attenuating caspase 3 and p53 mRNA activation. MEZA treatment also alleviated upregulation of α-SMA, type I collagen, and TGF-ß1 mRNA in the liver. The possible antifibrotic effect of MEZA was clearly demonstrated by the histopathology examination, using Masson's Trichrome and Sirius Red stainings. Therefore, the current study suggested that the bioactive compounds of Z. album possessed antifibrotic effect against DLM-induced hepatic fibrosis, by protecting liver tissue, and inhibiting oxidative stress, inflammation, apoptosis and the TGF-ß1/Smads signaling pathways.

(E)-N'-(1-(7-Hydroxy-2-Oxo-2H-Chromen-3-Yl) Ethylidene) Benzohydrazide, a Novel Synthesized Coumarin, Ameliorates Isoproterenol-Induced Myocardial Infarction in Rats through Attenuating Oxidative Stress, Inflammation, and Apoptosis.

The present study was directed to investigate the effect of precotreatment with (E)-N'-(1-(7-hydroxy-2-oxo-2H-chromen-3-yl) ethylidene) benzohydrazide (7-hyd.HC), a novel potent synthesized coumarin, on isoproterenol- (ISO-) induced myocardial infarction (MI) in rats. The hydrazone compound was characterized by IR, 1D, and 2D NMR analyses. Experimental induction of MI in rats was established by ISO (85 mg/kg/day, s.c) for two consecutive days (6th and 7th days). 7-hyd.HC or sintrom was given for 7 days prior and simultaneous to ISO injection. 7-hyd.HC offered a cardiopreventive effect by preventing heart injury marker leakage (LDH, ALT, AST, CK-MB, and cTn-I) from cardiomyocytes and normalizing cardiac function and ECG pattern, as well as improving lipid profile (TC, TG, LDL-C, and HDL-C), which were altered by ISO administration. Moreover, 7-hyd.HC precotreatment significantly mitigated the oxidative stress biomarkers, as evidenced by the decrease of lipid peroxidation and the increased level of the myocardial GSH level together with the SOD, GSH-Px, and catalase activities. 7-hyd.HC inhibited the cardiac apoptosis by upregulating the expression of Bcl-2 and downregulating the expression of Bax and caspase-3 genes. In addition, 7-hyd.HC reduced the elevated fibrinogen rate and better prevented the myocardial necrosis and improved the interstitial edema and neutrophil infiltration than sintrom. Overall, 7-hyd.HC ameliorated the severity of ISO-induced myocardial infarction through improving the oxidative status, attenuating apoptosis, and reducing fibrinogen production. The 7-hyd.HC actions could be mediated by its antioxidant, antiapoptotic, and anti-inflammatory capacities.

Zygophyllum album saponins prevent atherogenic effect induced by deltamethrin via attenuating arterial accumulation of native and oxidized LDL in rats.

The current study aimed to examine, for the first time, the relationship between exposure to deltamethrin (DLM) and atherogenic lipid profile disorders in adult Wistar rats, as well as, to verify the mechanism of the beneficial role of Zygophyllum album leaves extracts (ZALE). The experimental study was assessed using DLM (4 mg/kg b.w) either alone or co administered with ZALE (400 mg/kg b.w) orally for 90 days in rats. RP-HPLC-DAD-ESI-QTOF-MS was used to identify the bioactive metabolites present in ZALE. Plasmatic and aortic total cholesterol (TC), LDL-cholesterol (LDL-C), native LDL (LDL-apo B-100) and oxidized LDL (ox-LDL) were evaluated using auto-analyzer and a sandwich ELISA, respectively. The protein expressions of LDLR (native LDL receptor) and CD36 (Scavenger receptor class B) were evaluated in aorta or liver with a Western blot. The pathology has been confirmed with lipid stain (Oil Red O). Phytochemicals analysis revealed the presence of fifteen saponins in ZALE. Rats intoxicated with DLM revealed a significant increase in plasmatic and aortic lipid profile (TC, LDL-C, LDL-apo B-100 and ox-LDL), as well as, the concentration of the plasmatic cytokines include TNF-α, IL-2 and IL-6, compared to control. Hepatic native LDL and aortic CD36 receptor expression were increased in DLM treated group, however aortic LDL-R does not present any modification, when compared to control. The detected disturbances in lipid parameters were supported by Oil Red O applied. Due to their antioxidant activity, the bioactive compounds in ZALE as powerful agents able to prevent the pro-atherogenic effect observed in DLM-treated animals. These metabolites modulated most of inflammatory markers, prevented accumulation of lipid and lipoprotein biomarkers, regulated the major receptor regulators of hepatic cholesterol metabolism, as well as normalize lipid distribution in liver and aorta tissue.

Toxic effect of alpha cypermethrin, an environmental pollutant, on myocardial tissue in male wistar rats.

α-Cypermethrin (CYP) is a pyrethroid insecticide-like environmental pollutant, widely found in the environment. New research links exposure to high levels of CYP to health damage; however, little is known about the effect of CYP on cardiovascular disease. The purpose of the present study was to evaluate, for the first time, biochemical and cardiovascular changes in male rats resulting from subchronic CYP exposure. The animals were divided into three groups: group 1 served as the control, group 2 (CYP1) received 4 mg/kg of CYP by gavage, and group 3 (CYP2) received 8 mg/kg of CYP by gavage, for 8 weeks each. Results showed that both CYP1 and CYP2 markedly increased plasma concentrations of cardiac markers (LDH, CK-MB, and troponin-T). Moreover, compared to the control group, CYP treatment elevated cardiac oxidative stress, as shown by increased MDA level and decreased activity of SOD, CAT, and GSH-Px. In addition, CYP2 caused a significant increase of 42% the concentration of total cholesterol and more than 75% in triglycerides compared to the control group. Furthermore, DNA fragmentation and collagen deposition were both amplified owing to CYP toxicity. This harmful effect was confirmed by a histological study using H-E and Sirius Red staining. Overall, our results clearly proved the cardiotoxicity caused by α-cypermethrin.

HPLC-DAD-ESI-QTOF-MS/MS profiling of Zygophyllum album roots extract and assessment of its cardioprotective effect against deltamethrin-induced myocardial injuries in rat, by suppression of oxidative stress-related inflammation and apoptosis via NF-κB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Zygophyllum album is widely used to treat many cardiovascular diseases (CVDs) and as anti-inflammatory plant. AIM OF THE STUDY: This study aimed to investigate the mechanism of the potential protective effects of Zygophyllum album roots extract (ZARE) against myocardial damage and fibrosis induced by a chronic exposure to deltamethrin (DLM) in rats. MATERIALS AND METHODS: Bioactive compounds present in ZARE were analyzed by HPLC-DAD-ESI-QTOF-MS/MS. In vivo, DLM (4 mg/kg body weight), ZARE (400 mg/kg body weight) and DLM with ZARE were administered to rats orally for 60 days. Biochemical markers (LDH, ALT, CK, CK-MB and cTn-I) were assessed in the plasma by an auto-analyzer. Pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6) were evaluated by a sandwich ELISA. NF-κB was quantified at mRNA levels by real time PCR. Heart tissue was used to determine cardiac oxidative stress markers (MDA, PC, SOD, CAT, and GPx). Masson's Trichrome (MT) and Sirius Red (SR) stainings were used for explored fibrosis statues. RESULTS: Phytochemical analysis using HPLC-DAD-ESI-QTOF-MS/MS revealed the presence of twenty six molecules including phenolic compounds and saponins. ZARE significantly improved the heart injury markers (LDH, ALT, CK, CK-MB and cTn-I), lipid peroxidation (MDA), protein oxidation (PC), antioxidant capacity (SOD, CAT, and GPx), and DNA structure, which were altered by DLM exposure. Moreover, ZARE cotreatment reduced the expressions of NF-κB, decreased plasmatic pro-inflammatory cytokines concentration (TNF-α, IL-1ß and IL-6), and suppressed the myocardial collagen deposition, as observed by Sirius Red and Masson's Trichrome staining. CONCLUSION: ZARE ameliorated the severity of DLM-induced myocardial injuries through improving the oxidative status and reducing profibrotic cytokines production. The ZARE actions could be mediated by downregulation of NF-κB mRNA.

Bifenthrin exerts proatherogenic effects via arterial accumulation of native and oxidized LDL in rats: the beneficial role of vitamin E and selenium.

The purpose of this study was to investigate, for the first time, the effects of Bifenthrin (Bif) chronic exposure on plasmatic and aortic lipid parameters disturbance and their pro-atherogenic possibility in Wistar rats. The ameliorative role of vitamin E (Vit E) and selenium (Se) were also targeted. Thus, rats were treated by gastric gavage with combination of Vit E (100 mg/kg/bw) and Se (0.25 mg/kg/bw) in alone and co-treated groups for 90 days. Apart from control and Vit E-Se groups, all the groups were subjected to Bif (3 mg/kg, via gavage) toxicity. Results showed that Bif increased markedly plasmatic and aortic total cholesterol, LDL-cholesterol, native LDL-apoB-100, and oxidized-LDL, compared to the control. Moreover, Bif treatment significantly increased the plasmatic levels of the pro-inflammatory cytokines TNF-α, IL-2, and IL-6. In addition, the densitometric quantification of protein bands showed that the amount of hepatic native LDL-receptor protein decreased significantly in the intoxicated rats compared to the control group. The expression of arterial LDL receptors (LDLRs) and scavenger receptors (CD36) was amplified owing to Bif toxicity. This harmful effect was confirmed by histological study using Oil-Red-O staining. Owing to their antioxidant capacities, Vit E and Se have maintained all the changes in plasma and aorta lipids and prevented the pro-atherogenic effect observed in Bif-treated animals.

Cardioprotective effects of (E)-4-hydroxy-N'-(1-(3-oxo-3H-benzo[f]chromen-2-yl)ethylidene)benzohydrazide: a newly synthesized coumarin hydrazone against isoproterenol-induced myocardial infarction in a rat model.

The current study was carried out to evaluate the effect of pretreatment and co-treatment with a newly synthesized coumarin hydrazone, (E)-4-hydroxy-N'-(1-(3-oxo-3H-benzo[f]chromen-2-yl)ethylidene)benzohydrazide (hereinafter EK6), against isoproterenol-induced myocardial infarction in rats. Changes in biochemistry, cardiac biomarkers, electrocardiography, and histopathology after treatment with EK6 or acenocoumarol (Sintrom) were studied. Animals were randomly divided into 4 groups: vehicle control (C), isoproterenol + Sintrom (ISO + Sin), isoproterenol + EK6 (ISO + EK6), and isoproterenol (ISO). Myocardial infarction was induced by subcutaneous ISO administration at a dose of 85 mg·kg-1·day-1 with a drug-free interval of 24 h on days 6 and 7. Treatment with ISO led to significant elevation (p < 0.05) in serum levels of cardiac injury biomarkers, namely cardiac troponin-T, lactate dehydrogenase, creatine kinase-MB, alanine aminotransferase, and aspartate aminotransferase compared with levels in the vehicle control. A change in the lipid profile was also observed as a significant increase in total cholesterol and triglycerides. Furthermore, ISO caused significant alterations in the electrocardiogram pattern, including significant ST-segment elevation, significant decreased R wave amplitude, and significant increase in heart rate (16%) as well as marked changes in the histopathology of the heart tissue. Pretreatment and co-treatment with newly synthesized coumarin hydrazone restored all ISO-induced biochemical, lipid, cardiac, and histopathological changes in rats with myocardial infarction.

Contribution of adiponectin polymorphisms to the risk of coronary artery disease in a North-African Tunisian population.

BACKGROUND: Adiponectin, an adipocyte-derived protein, is known to play a key role in the processes leading to atherosclerosis and coronary artery disease (CAD) through its anti-atherogenic, anti-inflammatory, antioxidative, and anti-apoptotic properties. In the current study, we have studied the association of two single nucleotide polymorphisms (SNPs) +45 T>G (rs2241766) and +276 G>T (rs1501299) of the adiponectin gene with coronary artery disease (CAD) on an Arab/North-African population from Tunisia. METHODS: Subjects comprised 277 patients with angiographically demonstrated CAD and 269 age- and gender-matched control subjects. The adiponectin genotypes were performed by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP). The contribution of adiponectin variants to CAD was analyzed by haplotype and regression analysis. RESULTS: Adiponectin +45T>G and +276G>T genotypic and allelic distributions did not show a significant difference between cases and controls. Similarly, no association with CAD was observed for the haplotype analysis. Assuming dominant model of transmission for both polymorphisms and after adjustment of a number of traditional risk factors for CAD, logistic regression analysis showed an association of SNP +45 T>G with increased risk of developing CAD [adjusted OR (95% CI) = 2.59 (1.17-5.70); P = .01]. However, SNP + 276 G>T is associated with decreased risk of developing CAD [adjusted OR (95% CI) = 0.47 (0.22-0.97); P = .04]. CONCLUSION: There is no allelic or genotypic association of +45 T>G and +276 G>T of the adiponectin gene with CAD in the Tunisian population.

Persistent plasticizers and bisphenol in the cheese of Tunisian markets induced biochemical and histopathological alterations in male BALB/c mice.

Clinical evidences, experimental models, and epidemiology of many studies suggest that phthalate-based plasticizers, aliphatic ester, and bisphenol A (BPA) have major risks for humans by targeting different organs and body systems. The current study has been designed firstly to analyze three categories of cheese with and without their exposure to the sun and packed in packages with an inner surface plastic-covered film in order to identify the dibutyl phthalate (DBP); benzyl butyl phthalate (BBP); bis(2-ethylhexyl) phthalate (DEHP); diisodecyl phthalate (DiDP); diisononyl phthalate (DiNP); and 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) by GC-MS/MS, except for the bisphenol A, which is by UPLC-MS/MS, and secondly to assess the toxicity of the identified chemical molecules and cheese samples on the liver and kidney of mice. Our results showed that the cheese contains high quantities of DBP and DEHP with the concentrations up to 0.46 and 2.339 mg/kg, respectively. Other types of cheese, such as rolled and triangular cheeses, contain little quantities of the all substances at concentrations below the standard limits. In vivo, the obtained data clearly demonstrated that the acute administration of DBP, DEHP, and the tested cheese significantly induced liver and kidney injuries in mice manifested by a rise in plasma alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, lactate dehydrogenase, urea, creatinine, and uric acid when compared with control animals. In addition, the histopathological study confirmed the perturbation of biochemical parameters and showed that the hepatic and renal structures were altered. Indeed, the hepatotoxic and nephrotoxic effects are more pronounced when cheese was exposed to the sun.

Plasticizers and bisphenol A, in packaged foods sold in the Tunisian markets: study of their acute in vivo toxicity and their environmental fate.

Today, processed and packaged foods are considered as among the major sources of human exposure to plasticizers and bisphenol which migrate from plastic packing. In the present study, a wide range of food products sold on the Tunisian market such as grain and grain products, milk and dairy products, fats and oil, drink, fish, and sweets have been analyzed firstly in order to identify the presence of phthalates and bisphenol. Then, the identified chemical molecules were studied for their environmental fate and tested in vivo for its toxicity in mice models. The food products analyzed using GC-MS/MS indicated the presence of the benzyl butyl phthalate (BBP), dibutyl phthalate (DBP), bis(2-ethylhexyl) phthalate (DEHP), diisodecyl phthalate (DiDP), diisononyl phthalate (DiNP), and 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINC) and which using UPLC-MS/MS demonstrated the presence of bisphenol A of all food products. However, compared to other phthalates, BBP was found at high concentrations in the puff pastry (123 mg/kg), milk (2.59 mg/kg), butter (1.5 mg/kg), yogurt (2.23 mg/kg), oil (6.94 mg/kg), water (0.57 mg/kg), candy 1 (2.35 mg/kg), candy 2 (0.81 mg/kg), orange juice (1.25 mg/kg), peach juice (1.26 mg/kg), fruit juices (0.4 mg/kg), and chocolate (0.884 mg/kg). The obtained data in vivo clearly showed that the acute administration of BBP caused hepatic and renal damage as demonstrated by an increase in biochemical parameters as well as the activities of plasma marker enzymes such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, blood urea nitrogen, glucose, urea, creatinine, and uric acid when compared to the control group. By the same occurrence, the histopathological study revealed that BBP strongly modified the structure of hepatic and renal tissues. In addition, the plasticizers and BBP will therefore discharge via wastewater treatment plants in aquatic system and could reach marine organisms such as fish. We have followed the fate of BBP in bream Sparus aurata. In fact, chemical analysis showed the contamination of wild S. aurata by BBP from Sousse Coast (1.5 mg/kg) and wild S. aurata from Monastir Coast (0.33 mg/kg).

Cytotoxic effects of seven Tunisian hospital wastewaters on the proliferation of human breast cancer cell line MDA-231: correlation with their chemical characterization.

Hospital wastewaters contain large amounts of pharmaceutical residues, which may eventually be discharged into the aquatic environment through wastewater treatment plants, raising the question of their impact on human and environmental health. This has prompted the launch of several monitoring studies into the most commonly administered compounds in urban wastewater. The aim of this study was, therefore, to explore the cytotoxic potential of wastewaters samples collected from seven hospitals in Tunisia. The physicochemical analyses showed a large fluctuation of certain parameters in the collected samples, such as chemical oxygen demand (ranged from 860 to 1720 mg L-1), biochemical oxygen demand (ranged from 385 to 747 mg L-1), total organic carbon (ranged from 256 to 562 g L-1), total suspended solids (ranged from 905 to 1450 mg L-1), conductivity (ranged from 3.31 to 7.14 µsm/cm), and turbidity (ranged from 100 to 480 NTU). The analysis using inductively coupled plasma mass spectrometry (ICP-MS) also showed that hospital wastewater contains high concentrations of Hg (ranged from 0.0024 to 0.019 mg L-1). This could be explained by the variation of the activity and the services in certain hospitals compared to others. All hospital wastewater samples induced the proliferation of human breast cancer cell line MDA-231, even at low concentrations (20 µL/assay). Moreover, the maximum induction reached at the concentration of 60 µL/assay in wastewater samples from hospitals located in Monastir, Sidi Bouzid, Mahdia, and Sfax with percentages of induction up to 42.33, 14, 7.61, and 5.42%, respectively. These observations could be due to the presence of endocrine disrupting compounds (EDCs) in these wastewaters. Given this, our results evidenced the potential risk of these hospital effluents to environmental and public health.

-174G>C interleukin-6 gene polymorphism in Tunisian patients with coronary artery disease.

BACKGROUND AND OBJECTIVES: A state of low-grade inflammation accompanies the pathogenesis of atherosclerotic events. Interleukin-6 (IL-6) is a pleotropic pro-inflammatory cytokine that modulates the development of acute coronary syndromes (ACSs), partly by destabilizing coronary atherosclerotic plaques. We have examined the contribution of the -174G>C IL-6 promoter variant on the risk of coronary artery disease (CAD) among Tunisians. PATIENTS AND METHODS: Study subjects included 418 CAD patients and 406 age- and sex-matched controls. IL-6 genotyping was done by PCR-restriction fragment length polymorphism. RESULTS: The frequency of the -174C allele (mutant) was lower in Tunisians than in Europeans, and the distribution of -174 G>C genotypes was similar between CAD patients and control subjects. Moreover, compared to GG genotype carriers, -174C allele carriage did not increase the CAD relative risk (odds ratio and 95% confidence interval=1.09 and 0.80-1.49), which remained nonsignificant after adjusting for traditional risk factors for CAD (age, smoking, hypertension, diabetes and obesity). CONCLUSION: The -174G>C IL-6 promoter variant is not associated with an increased risk of CAD among Tunisians.

Contribution of SELP and PSGL-1 genotypes and haplotypes to the presence of coronary heart disease in Tunisians.

P-selectin (SELP) and its counter-receptor, P-selectin glycoprotein ligand-1 (PSGL-1), play key role in the transient attachment of leukocytes to endothelial cells predisposing to coronary heart disease (CHD). In the current report, 293 angiographically proven CHD patients and 327 age, gender, and race-matched controls were included. Our aim was to evaluate the contribution to CHD of the following SNPs: C-2123G, G-1969A and T715P in SELP, Met62Ile and the VNTR variants in PSGL-1 gene in a North African population from Tunisia. While there were no significant differences in the distribution of SELP or PSGL-1 alleles or genotypes between patients and controls, a trend for a significant association of the C-2123G genotypes distribution with incident CHD was observed (P=0.06). Assuming an additive model of transmission, the risk was 74% higher among subjects carrying the GG genotypes in comparison to those carrying the CC genotype (OR=1.74 [1.01-2.98], P=0.04) and 80% higher in the recessive model (OR=1.80 [1.08-3.01], P=0.02). Haplotype analysis did not identify any specific SELP or PSGL-1 haplotypes to be associated with CHD. The present study demonstrated no evidence of association between individual SELP or PSGL-1 SNPs or haplotypes with incident CHD. However, this study replicates absence of association of the mostly studied SNP, T715P, previously reported in individuals with African origin.

Functional interleukin-10 promoter variants in coronary artery disease patients in Tunisia.

OBJECTIVES: The contribution of interleukin (IL)-10 promoter variants -1082G/A, -819C/T, and -592C/A to the risk of coronary artery disease (CAD) was investigated in 291 CAD patients and 291 age- and gender-matched control subjects. METHODS AND RESULTS: IL-10 genotyping was performed using PCR-allele-specific amplification (PCR-ASA). Regression analysis was employed in assessing the contribution of the IL-10 variants to the overall CAD risk. A higher frequency of the -592A allele (p = 0.004), but not the -1082A (p = 0.828) or -819T (p = 0.952) alleles, was seen in CAD patients. A higher frequency of -592C/A (p = 0.011), and a lower frequency of -592C/C (p = 0.015) genotypes was noted in patients compared to healthy controls. Regression analysis demonstrated an association of -592C/A [OR (95% CI) = 1.82 (1.02-3.23)] and -592A/A [OR (95% CI) = 3.33 (1.27-9.09)] genotypes with 1-artery disease. Haplotype analysis revealed that none of the eight possible IL-10 haplotypes was associated with CAD or with the severity of CAD, and was confirmed by multivariate regression analysis, after adjusting for a number of confounders (smoking, systolic and diastolic blood pressure, hypertension, diabetes, glucose, cholesterol, and triglycerides). CONCLUSIONS: Our results suggest that the -592C/A, more so than the -1082G/A or the -819C/T IL-10 promoter variant alleles, may be considered to be a risk factor for CAD in Tunisians.