Wild Vitex agnus-castus L.: Phytochemical Characterization, Acute Toxicity, and Bioactive Properties.

Wild Vitex agnus-castus (VAC) is a Mediterranean plant that is rich in bioactive metabolites. This study aimed to validate, for the first time, the beneficial use of VAC fruits and fruit decoctions (VFDs) through in vitro and in vivo trials. Forty-one volatile components were detected in VAC fruits, with 1,8-cineole (30.3%) comprising the majority. The antioxidant activity of VFD was measured by using different in vitro methods (EC50 of 0.16 mg/mL by ß-carotene bleaching inhibition assay) and by measuring the DNA protection power. Using the disc diffusion assay, the antimicrobial activity of VFD was evaluated, and it exhibited a noticeable anticandidal activity. VFD did not cause any toxicity or mortality in rats treated with doses > 200 mg/kg. Using the acetic acid writhing test, the antinociceptive activity of VFD was measured. Our results showed that VFD at 200 mg/kg exhibited a higher analgesic activity (81.68%) than acetylsalicylic acid used as a positive control (74.35%). Its gastroprotective ability was assessed by HCl/ethanol-induced gastric lesions, which were remarkably inhibited (84.62%) by intraperitoneal administration of VFD. This work helps to validate the popular use of VAC to treat nociceptive, inflammatory, and gastric disorders and encourages researchers to further investigate the identification of pharmacological compounds from this species.

Effect of Flavoring with Rosemary, Lemon and Orange on the Quality, Composition and Biological Properties of Olive Oil: Comparative Study of Extraction Processes.

The goal of this work was to investigate the impact of the flavoring of some aromatic plants/spices, including rosemary (R), lemon (L) and orange (O) at the concentration of 5% and 35% (w/w) added by 2 methods (conventional maceration and direct flavoring), on quality attributes, chemical changes and oxidative stability of extra virgin olive oil (EVOO). Six flavored oils were obtained (EVOO + O, O + O, EVOO + R, O + R, EVOO + L and O + L). The physicochemical parameters (water content, refractive index, acidity and peroxide value, extinction coefficient, fatty acids, volatile aroma profiles, Rancimat test, phenols and pigments composition) of the flavored oils were investigated. Based on the results obtained, it was observed that flavoring with a conventional process provided increased oxidative stability to the flavored oils, especially with rosemary (19.38 ± 0.26 h), compared to that of unflavored oil. The volatile profiles of the different flavored oils revealed the presence of 34 compounds with the dominance of Limonene. The fatty acid composition showed an abundance of mono-unsaturated fatty acids followed by poly-unsaturated ones. Moreover, a high antioxidant activity, a significant peripheral analgesic effect (77.7% of writhing inhibition) and an interesting gastroprotective action (96.59% of ulcer inhibition) have been observed for the rosemary-flavored oil. Indeed, the flavored olive oils of this study could be used as new functional foods, leading to new customers and further markets.

Chemical characterization and nutritional quality investigations of healthy extra virgin olive oil flavored with chili pepper.

The production of extra virgin olive oil (EVOO) flavored with diverse spices, herbs, fruits, and vegetables or natural aromas is believed to provide advantageous properties considering either the high nutritional value or biological activity in addition to the flavoring and industrial aspects. The biological activities including antioxidant and antimicrobial properties of Tunisian EVOO obtained from "Chemlali" variety and mixed with chili pepper were investigated. Molecular analyses, including the detection of twelve olive-infecting viruses and Pseudomonas savastanoi pv savastanoi, were performed to ensure that the samples were obtained from healthy olive trees and EVOO quality was not affected. Quality parameters like free acidity, peroxide number, oxidative stability, and specific absorption at K232 nm and K270 nm were also investigated and no significant variation was revealed. The content of minor compounds such as chlorophylls, carotenoids, and total phenols showed minor changes. However, the profiles of the volatile compounds showed remarkable differences, which appeared to be the main factor for the observed variability in consumer acceptance. The results showed for the first time high quantities of polyphenols and ortho-diphenols. Four colorimetric methods were used for the determination of the antioxidant activity, namely DPPH, ABTS, FRAP, and ß-carotene test. Compared to the control, a higher level of antioxidant activity was observed for the flavored EVOO. Furthermore, significant results were obtained in the antimicrobial tests. The quality parameters of the mixture showed no alteration compared to the control. Finally, all the measurements and the chemical characterization gave a scientific basis for food technology innovation of new food products.

Effect of oleuropein on oxidative stress, inflammation and apoptosis induced by ischemia-reperfusion injury in rat kidney.

AIMS: This study aimed to evaluate the effect of oleuropein (OLE), the main phenolic compound present in olive leaves, on kidney ischemia-reperfusion injury (IRI) and to explore the underlying protective mechanism. MAIN METHODS: Rat kidneys were subjected to 60 min of bilateral warm ischemia followed by 120 min of reperfusion. OLE was administered orally 48 h, 24 h and 30 min prior to ischemia at doses of 10, 50 and 100 mg/kg body weight. The creatinine, urea, uric acid concentrations and lactate dehydrogenase (LDH) activity in plasma were evaluated. Oxidative stress and inflammation parameters were also assessed. Renal expression of AMP-activated protein kinase (p-AMPK), endothelial nitric oxide synthase (eNOS), mitogen-activated protein kinases (MAPK), inflammatory proteins and apoptotic proteins were evaluated using Western blot. KEY FINDINGS: Our results showed that OLE at 50 mg/kg reduced kidney IRI as revealed by a significant decrease of plasmatic creatinine, urea, uric acid concentrations and LDH activity. In parallel, OLE up-regulated antioxidant capacities. Moreover, OLE diminished the level of CRP and the expression of cyclooxygenase 2 (COX-2). Finally, OLE enhanced AMPK phosphorylation as well as eNOS expression whereas MAPK, and cleaved caspase-3 implicated in cellular apoptosis were attenuated in the ischemic kidneys. SIGNIFICANCE: In conclusion, this study shows that OLE could be used as therapeutic agent to reduce IRI through its anti-oxidative, anti-inflammatory and anti-apoptotic properties.

Nutritional properties, identification of phenolic compounds, and enzyme inhibitory activities of Feijoa sellowiana leaves.

The nutritional values, the identification of individual phenolic compounds as well as the functional properties (in vitro acetylcholinesterase (AChE) and pancreatic lipase inhibition) of Tunisian Feijoa sellowiana (O.Berg) leaves were determined by high-performance liquid chromatography coupled with diode array and mass spectrometry detector. Feijoa Sellowiana  leaves Extract (FSLE) were evaluated in the present work. Macronutrient components (proteins, carbohydrates, dietary fiber, lipids, and ashes) of FSLE as well as their fatty acids and mineral content were determined. The individual phenolic composition was characterized by the presence of flavonols (mostly quercetin glycosides), flavan-3-ols (procyanidins and catechins), and phenolic acid derivatives. To understand the possible inhibition mechanism exerted by the extract components, enzyme kinetic studies using derived Lineweaver-Burk (LB) plots and Michaelis-Menten were performed. The obtained results evidenced that FSLE exhibited important AChE inhibitory effect, behaving as a noncompetitive inhibitor. Furthermore, the leaves' extract inhibited irreversibly the pancreatic lipase, with maximum inhibition (70%) achieved at 1 mg/ml. PRACTICAL APPLICATIONS: In many countries of the world, Feijoa Sellowiana leaves are used as an alternative to tea drinks and as a folkloric medicine. Moreover, it is well known that the phytochemical compounds are a practical alternative for the prevention of many chronic diseases such as Alzheimer's and obesity. For this reason, the present study aimed to evaluate the nutritional, individual phenolic compounds, and functional properties of Feijoa Sellowiana leaves to valorize them as a food and pharmaceutical bioactive human source.

Cytoskeleton involvement in lithium-induced SH-SY5Y neuritogenesis and the role of glycogen synthase kinase 3ß.

Lithium modulates signals impacting on the cytoskeleton, a dynamic system contributing to neural plasticity at multiple levels. In this study, SH-SY5Y human neuronal cells were cultured in the absence (C) or in presence (Li) of a 0.5 mM Li2CO3 (i.e. 1 mM lithium ion) for 25-50 weeks. We investigated the effect of this treatment on (1) morphological changes of cells observed using Hemalun eosin staining assay, (2) cytoskeletal changes by indirect immunofluorescence (IIF) staining of microtubules (α-tubulin) and heavy neurofilaments subunits (NF-H) and by measuring the expression rate changes of genes coding for receptor for activated C kinase (RACK1), casein kinase2 (CK2) and thymosine beta-10 using cDNA arrays technology, (3) cell adhesion properties by IIF staining of ß-catenin protein. Besides, we have tried to understand the molecular mechanism of lithium action that triggers changes in cytoskeleton and neurites outgrowth. Thus, we examined the effect of this treatment on glycogen synthase kinase 3 (GSK3) expression and activity using western blotting of GSK3 and phosphorylated ß-catenin, a downstream GSK3 target protein. Our results showed that lithium treatment reduces axon length, increases axonal spreading, enhances neurites growth and neurites branching with an increase of growth cone size. Moreover, genes coding for CK2 and thymosine beta-10 were significantly up-regulated, however, that coding for RACK1 was down-regulated. The most interesting result in this work is that mechanism underlying lithium action was not related to the inhibition of GSK3 activity. In fact, neither expression rate nor activity of this protein was changed.

Hematotoxicity and genotoxicity of mercuric chloride following subchronic exposure through drinking water in male rats.

Erythrocytes are a convenient model to understand the subsequent oxidative deterioration of biological macromolecules in metal toxicities. The present study examined the variation of hematoxic and genotoxic parameters following subchronic exposure of mercuric chloride via drinking water and their possible association with oxidative stress. Male rats were exposed to 50 ppm (HG1) and 100 ppm (HG2) of mercuric chloride daily for 90 days. A significant dose-dependent decrease was observed in red blood cell count, hemoglobin, hematocrit, and mean cell hemoglobin concentration in treated groups (HG1 and HG2) compared with controls. A significant dose-dependent increase was observed in lipid peroxidation; therefore, a significant variation was found in the antioxidant enzyme activities, such as superoxide dismutase, catalase, and glutathione peroxidase. Interestingly, mercuric chloride treatment showed a significant dose-dependent increase in frequency of total chromosomal aberration and in percentage of aberrant bone marrow metaphase of treated groups (p < 0.01). The oxidative stress induced by mercury treatment may be the major cause for chromosomal aberration as free radicals lead to DNA damage. These data will be useful in screening the antioxidant activities of natural products, which may be specific to the bone marrow tissue.

Impairment of spermatogenesis in rats by mercuric chloride: involvement of low 17ß-estradiol level in induction of acute oxidative stress.

Mercuric chloride (HgCl(2)) has been shown to affect the male reproductive organs, and oxidative stress has been linked with hypospermatogenesis and with male infertility. However, the specific mode of impairment of spermatogenesis during HgCl(2) exposure has not yet been clarified fully. Because of the involvement of 17ß-estradiol (E2) in the male reproductive tract and its putative role on spermatogenesis, the present study aimed to investigate the possibility that HgCl(2)-induced oxidative stress-mediated modulation of the E2 level exerts adverse effects on testicular steroidogenic and gametogenic activities. HgCl(2) treatment at 50 and 100 ppm for 90 days by continuous oral administration in the drink water resulted in significant dose-dependent fashion decrease in serum and testicular E(2) levels and an increase in testicular testosterone levels in dose-dependent manner, without statistical alteration in serum testosterone level among HgCl(2) exposed groups compared to the control. Cauda epididymal sperm count and motility were decreased significantly (p < 0.01), in a dose-dependent manner, in the HgCl(2)-treated groups, and qualitative examination revealed inhibition of spermatogenesis and the preferential loss of maturing and elongated spermatids. The seminiferous tubules were dilated in treated animals. When compared to the control, increase in lipid peroxidation due to toxic effects of HgCl2 was accompanied by significant reduction (p < 0.01) in antioxidant enzymes activities, superoxide dismutase, catalase, and glutathione peroxidase of testes, implicating the presence of oxidative tissue damage. Furthermore, these tissue injuries caused functional impairment as evidenced with testicular elevated activity of lactate dehydrogenase. Unless oxidative stress can lead to cancer development, testis' tumor markers as beta human chorionic gonadotropin and alpha-fetoprotein levels have shown no significant differences in the HgCl(2)-exposed group compared with respect to the control. Large quantities of metal accumulated in the testis tissue are in agreement with the testis-activity failure verified in this tissue. These findings suggest that a decrease in E2 level after mercury exposure may render testis more susceptible to oxidative damage leading to its functional inactivation, thus providing new dimension to mechanisms underlying heavy metal-induced male infertility.

Testicular toxicity in mercuric chloride treated rats: association with oxidative stress.

Mercury has been recognized as an industrial hazard that adversely affects male reproductive systems of humans and animals. However, less information is available concerning the underlying mechanism in the pathogenesis of male reproductive dysfunction. The present study investigated the possible involvement of oxidative stress to induce oxidative deterioration of testicular functions in adult rats. Wistar male rats (n=132) were continuously exposed to HgCl(2) at 0, 50 and 100 ppm during 90 days through oral administration in the drinking water. Mercury exposure for 90 days resulted in an increase in the absolute and relative wet weight of the testis and a decrease in the absolute and relative wet weight of the accessory sex glands, with respect to the matched control. Marked perturbation in testosterone serum level was also detected during the treatment for the treated groups. Cauda epididymal sperm count/motility decreased significantly in the mercury-treated group and qualitative examination of testicular sections revealed a fewer mature luminal spermatozoa in comparison to the control. When the mercury-treated males were mated with normal cyclic females, mercury exposure resulted in a decline of the reproductive performance of male rats. These effects were associated with a significant increase in mercury content of testes and blood in time-dependent and dose-dependent fashion, respectively. The HgCl(2) treatment was associated with oxidative stress. Evidence of induction of oxidative stress was obtained in terms of perturbations in antioxidant defense and a significant dose-dependent increase in the testicular lipid peroxidation as a consequence of pro-oxidant exposure. Taken together, the results suggest that an increase in free radical formation relative to loss of antioxidant defense system after mercury exposure may render testis more susceptible to oxidative damage leading to their functional inactivation.

Combined vitamins (C and E) and insulin improve oxidative stress and pancreatic and hepatic injury in alloxan diabetic rats.

The aim of the present study is to determine if a combination of vitamins (C and E) has any advantage over insulin therapy on lipid peroxidation, antioxidant activity, liver dysfunction parameters, and histological changes in the alloxan-induced diabetic rats. The enzymatic activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT) and the lipid peroxidation product, thiobarbituric acid-reacting substances (TBARS) were measured in liver and pancreas as indicators of antioxidation in these tissues. The liver dysfunction parameters: the activity of lactate dehydrogenase (LDH), gamma glutamyl transferase (GGT), phosphatase alkalines (PAL), aspartate and lactate transaminase (AST and ALT) were measured in serum. In diabetic rats, the TBARS contents of the liver and pancreatic tissues were found to have significantly increased as compared to non-diabetic rats (P < 0.001). The SOD, CAT, and GPX activities in the liver and pancreas in diabetic rats significantly decreased as compared to normal rats (P < 0.001). AST, ALT, LDH, GGT, and PAL activities increased in the diabetic rats (p > 0.05). In diabetic rats treated with insulin or with combined vitamins (C and E), an ameliorative effect was observed. This amelioration was more pronounced in the group of rats treated with combined vitamins (C and E).

Hyperglycaemia, stress oxidant, liver dysfunction and histological changes in diabetic male rat pancreas and liver: protective effect of 17 beta-estradiol.

Oxidative stress is thought to play a crucial role in the pathogenesis of chronic diabetic complications. We investigated the protective effects of 17 beta-estradiol (E2) on alloxan-induced stress oxidant, hepatic dysfunction and histological changes in male rats liver and pancreas. Our results showed that 17 beta-estradiol could attenuate the increase of blood glucose in plasma and normalise the hepatic glycogen level. In addition, E2 enhanced superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) (by 207, 52 and 72%, respectively, as compared to diabetic rats), reduced lipid peroxidation in the hepatic tissue (by 54%) and improved the liver dysfunction parameters by the significant decrease of gamma-glytamyl transferase (GGT), phosphatases alkalines (PAL), lactate deshydrogenase (LDH) and aspartate and lactate transaminases (AST and ALT) activities which increased in diabetic rats. Moreover, 17 beta-estradiol treatment in diabetic rats protects against alloxan-induced pancreatic beta-cells and hepatic cells damages.