Perineural invasion in incisional biopsy is an indication for in-continuity resection in oral squamous cell carcinoma of the tongue and floor of the mouth.

PURPOSE: The histologic finding of perineural invasion is a poor prognostic factor in oral squamous cell carcinoma patients. The aim of this study is to detect factors that influence prognosis in patients operated for oral cancer with perineural invasion in their histopathology report and to propose safer options of treatment with respect to recurrence and survival rates. PATIENTS AND METHODS: We performed a prospective cohort study of all oral squamous cell carcinoma cases operated in our department from January 2018 to January 2022. Overall, 75 cases of patients operated for oral squamous cell carcinoma were included in the study. Of these cases, 14 with perineural invasion were found. We created two groups: one group of patients treated with in-continuity resection and a second group of patients treated with a discontinuous resection between the primary site and the neck. Type of operation was considered the predictor variable and recurrence, survival or death, the outcome variables. RESULTS: Oral cancer patients with known perineural invasion treated with in-continuity resection had a much better overall and disease-specific survival than patients treated with a discontinuous manner (p < .001). The difference in recurrence rate between these two groups of patients was found to be very significant in our study (p < .0005). CONCLUSION: It is therefore our suggestion that in oral squamous cell carcinoma of the tongue and floor of the mouth with perineural invasion detected in the incisional biopsy, an extended in-continuity resection to include the affected nerves should be performed.

The Bioactivity of a Hydroxytyrosol-Enriched Extract Originated after Direct Hydrolysis of Olive Leaves from Greek Cultivars.

Nowadays, olive leaf polyphenols have been at the center of scientific interest due to their beneficial effects on human health. The most abundant polyphenol in olive leaves is oleuropein. The biological properties of oleuropein are mainly due to the hydroxytyrosol moiety, a drastic catechol group, whose biological activity has been mentioned many times in the literature. Hence, in recent years, many nutritional supplements, food products, and cosmetics enriched in hydroxytyrosol have been developed and marketed, with unexpectedly positive results. However, the concentration levels of hydroxytyrosol in olive leaves are low, as it depends on several agricultural factors. In this study, a rapid and easy methodology for the production of hydroxytyrosol-enriched extracts from olive leaves was described. The proposed method is based on the direct acidic hydrolysis of olive leaves, where the extraction procedure and the hydrolysis of oleuropein are carried out in one step. Furthermore, we tested the in vitro bioactivity of this extract using cell-free and cell-based methods, evaluating its antioxidant and DNA-protective properties. Our results showed that the hydroxytyrosol-enriched extract produced after direct hydrolysis of olive leaves exerted significant in vitro antioxidant and geno-protective activity, and potentially these extracts could have various applications in the pharmaceutical, food, and cosmetic industries.

Poorly controlled diabetes mellitus is strongly associated with descending necrotizing mediastinitis of odontogenic origin.

OBJECTIVE: We hypothesized that poorly controlled diabetic status may be a negative prognostic factor for acute mediastinitis of odontogenic origin, and we conducted a cohort study to verify this hypothesis. STUDY DESIGN: Data were collected on all consecutive patients diagnosed with maxillofacial infections of odontogenic origin who received surgical treatment in the Oral and Maxillofacial Surgery Department. All patients included in the study were divided into 4 groups based on the presence or absence of diabetes and mediastinitis. Poorly controlled diabetic status constituted our primary predictor variable, and the occurrence of descending necrotizing mediastinitis was the outcome variable. RESULTS: All 7 patients with poorly controlled diabetes mellitus were complicated with descending necrotizing mediastinitis. An exact Fischer test was conducted to investigate the association of poorly controlled diabetes mellitus with descending necrotizing mediastinitis in patients admitted for surgical treatment of their maxillofacial collection. The result was significant at P < .01. CONCLUSION: This study represents the first scientific attempt to associate poorly controlled diabetes mellitus with descending necrotizing mediastinitis of odontogenic origin. Our results show evidently that these patients are more susceptible to deep space infections, which subsequently spread faster and are much more difficult to control.

Exploring the Bioactive Content of Liquid Waste and Byproducts Produced by Two-Phase Olive Mills in Laconia (Greece): Is There a Prospect for Added-Value Applications?

The use of a two-phase decanter (TwPD) for olive-oil extraction produces wastes and byproducts (a small volume of water from oil washing, olive leaves from the defoliator, and a high moisture pomace which can be destoned) that contain valuable bioactive compounds, such as phenolics and/or triterpenic acids. So far, there is no (water) or limited information (leaves and the destoned pomace fraction) on their content of bioactives, especially triterpenic acids. To contribute to the characterization of such streams from cultivars of international interest, in the present study, samples obtained from five mills from the region of Laconia (from one or two harvests) in Greece, where Koroneiki cv dominates, were screened for phenols and/or triterpenic acids. The leaves and pomace were dried at two temperatures (70 °C and/or 140 °C), and the pomace was also destoned before analysis. The liquid wastes contained low amounts of total (TPC) phenols (<140 mg gallic acid/L), hydroxytyrosol (<44 mg/L), and tyrosol (<33 mg/L). The olive leaves varied widely in TPC (12.8-57.4 mg gallic acid/g dry leaf) and oleuropein (0.4-56.8 mg/g dry leaf) but contained an appreciable amount of triterpenic acids, mainly oleanolic acid (~12.5-31 mg/g dry leaf, respectively). A higher drying temperature (140 vs. 70 °C) affected rather positively the TPC/oleuropein content, whereas triterpenic acids were unaffected. The destoned pomace TPC was 15.5-22.0 mg gallic acid/g dw, hydroxytyrosol 3.9-5.6 mg/g dw, and maslinic 5.5-19.3 mg/g dw. Drying at 140 °C preserved better its bioactive phenols, whereas triterpenic acids were not influenced. The present findings indicate that TwPD streams may have a prospect as a source of bioactives for added-value applications. Material handling, including drying conditions, may be critical but only for phenols.

Heat Shock Proteins' Expression Profiles in Odontogenic Ameloblastomas and Cysts.

Tumors and cysts with odontogenic origin represent a family of lesions with specific histo-genetic and clinical characteristics. Among them, ameloblastomas are common benign neoplasms, predominantly detected in the anatomic areas of the jaws and also in the mandible and maxilla. Although they are characterized by a slow and stable growing pattern, a subset of them shows a tendency for local tissue invasiveness and partially increased recurrence rates after surgical excision. Furthermore, heat shock proteins (HSPs) are potentially implicated in ameloblastoma onset and progression. HSPs regulate the folding and refolding of proteins and are induced in response to oxidative stress. They are crucial members of the chaperone intracellular system and are categorized based on their molecular weight (i.e., HSP27, HSP60, HSP70, HSP90). In the current review, we describe HSPs origin and function, focusing on their deregulation mechanisms and impact predominantly on ameloblastomas and also on inflammatory and developmental odontogenic cystic lesions.

Effects of Enriched-in-Oleuropein Olive Leaf Extract Dietary Supplementation on Egg Quality and Antioxidant Parameters in Laying Hens.

The objective of the present study was to evaluate the effects of an olive leaf extract obtained with an up-to-date laboratory method, when supplemented at different levels in laying hens' diets, on egg quality, egg yolk antioxidant parameters, fatty acid content, and liver pathology characteristics. Thus, 96 laying hens of the ISA-Brown breed were allocated to 48 experimental cages with two hens in each cage, resulting in 12 replicates per treatment. Treatments were: T1 (Control: basal diet); T2 (1% olive leaf extract); T3 (2.5% olive leaf extract); T4 (Positive control: 0.1% encapsulated oregano oil). Eggshell weight and thickness were improved in all treatments compared to the control, with T2 being significantly higher till the end of the experiment (p < 0.001). Egg yolk MDA content was lower for the T2 and T4 groups, while total phenol content and Haugh units were greater in the T2. The most improved fatty acid profile was the one of T3 yolks. The α-tocopherol yolk content was higher in all groups compared to T1. No effect was observed on cholesterol content at any treatment. Based on the findings, it can be inferred that the inclusion of olive leaf extract at a concentration of 1% in the diet leads to enhancements in specific egg quality attributes, accompanied by an augmentation of the antioxidant capacity.

Antioxidant and DNA-Protective Activity of an Extract Originated from Kalamon Olives Debittering.

Table olives are a major component of the Mediterranean diet and are associated with many beneficial biological activities, which are mainly related to their phenolic compounds. Olive fruit debittering process defines the quantitative and qualitative composition of table olives in biophenols. The aim of the present study was to evaluate the in vitro antioxidant capacity and DNA-protective activity of an extract originated from brine samples, according to the Greek style debbitering process of Kalamon olive fruits. The main phenolic components determined in the brine extract were hydroxytyrosol (HT), verbascoside (VERB) and tyrosol (T). The in vitro cell-free assays showed strong radical scavenging capacity from the extract, therefore antioxidant potential. At cellular level, human endothelial cells (EA.hy296) and murine myoblasts (C2C12) were treated with non-cytotoxic concentrations of the brine extract and the redox status was assessed by measuring glutathione (GSH), reactive oxygen species (ROS) and lipid peroxidation levels (TBARS). Our results show cell type specific response, exerting a hormetic reflection at endothelial cells. Finally, in both cell lines, pre-treatment with brine extract protected from H2O2-induced DNA damage. In conclusion, this is the first holistic approach highlighted table olive wastewaters from Kalamon- Greek style debittering process, as valuable source of bioactive compounds, which could have interesting implications for the development of new products in food or other industries.

Direct In Vitro Comparison of the Anti-Leishmanial Activity of Different Olive Oil Total Polyphenolic Fractions and Assessment of Their Combined Effects with Miltefosine.

The bioactive compounds present in the edible products of the olive tree have been extensively studied and their favorable effects on various disease risk factors have been demonstrated. The aim of this study was to perform a comparative analysis of the anti-leishmanial effects of total phenolic fractions (TPFs) derived from extra virgin olive oil with different phenolic contents and diverse quantitative patterns. Moreover, the present study investigated their association with miltefosine, a standard anti-leishmanial drug, against both extracellular promastigotes and intracellular amastigotes of a viscerotropic and a dermotropic Leishmania strain. The chemical compositions of TPFs were determined by high performance liquid chromatography with diode array detection (HPLC-DAD). Analysis of parasite growth kinetics, reactive oxygen species production and apoptotic events were determined by microscopy and flow cytometry. Our results revealed that the presence of oleacein (OLEA) and oleocanthal (OLEO) secoiridoids enhances the anti-leishmanial effect of TPF. The association between TPFs and miltefosine was suggested as being additive in Leishmania infantum and Leishmania major promastigotes, and as antagonistic in intracellular amastigotes, as was evaluated with the modified isobologram method. The obtained data verified that TPFs are bioactive dietary extracts with a strong anti-leishmanial activity and highlighted that fractions that are richer in OLEA and OLEO phenolic compounds possess stronger inhibitory effects against parasites. This study may contribute to improving the therapeutic approaches against leishmaniasis.

Higher Yield and Polyphenol Content in Olive Pomace Extracts Using 2-Methyloxolane as Bio-Based Solvent.

Despite its severe toxicity and negative environmental impact, hexane remain the solvent of choice for the extraction of vegetable oils. This is in contrast with the constantly growing demand for sustainable and green extraction processes. In recent years a variety of alternatives to hexane have been reported, among them 2-methyloxolane (2-MeOx), which has emerged as a promising bio-based alternative. This study evaluates the possibility of replacing hexane, in the extraction of olive pomace (OP), with 2-MeOx, both dry and saturated with water (4.5%), the latter of which is called 2-MeOx 95.5%. The three solvents have been compared in terms of extraction yield and quality, as well as the lipid and polyphenol profiles of the extracts. The work concluded that both dry 2-MeOx and 2-MeOx 95.5% can replace hexane in OP extraction, resulting in higher yields and extracts richer in phenolic compounds. This study should open the road to further semi-industrial scale investigations toward more sustainable production processes.

Oxidized Forms of Olive Oil Secoiridoids: Semisynthesis, Identification and Correlation with Quality Parameters.

Secoiridoids is the prominent chemical class of olive oil polar constituents and are characterized by significant biological properties. They are abundant in different chemical forms and relatively high concentrations compared to other components, while prone to oxidation due to their chemical motif. In recent years, oxidized derivatives of secoiridoids have been reported, either as natural constituents of olive oil or as components which are gradually formed in all stages of its production and storage. The mono-oxidized forms of oleocanthal and oleacein named as the respective acids have been recently isolated from olive oil and unambiguously structurally characterized. Other oxidized forms of elenolic acid or more complex secoiridoids, such as those of oleuropein and ligstroside aglycones are also sporadically mentioned in the literature. No further information is provided since they have not been isolated in pure form in order to be accurately identified. Most of the time, they are generally referred as oxidized forms of the parent compounds and commonly identified based on mass spectrometric data. In the current study, the semi-synthesis of the main oxidized olive oil secoiridoids, i.e., oleocanthalic acid, oleaceinic acid, EDA acid, carboxylic form of elenolic acid, carboxylic form of ligstroside aglycon, and carboxylic form of oleuropein aglycon is described starting from the corresponding aldehydic derivatives, using SeO2/H2O2 as oxidative agents. Furthermore, their presence in a number of Greek olive oils was investigated as well, as possible correlation thereof with quality parameters.

Intensification of Biophenols Extraction Yield from Olive Pomace Using Innovative Green Technologies.

Olive pomace is the main by-product generated by the olive oil production process. Although toxic to the environment, olive pomace is an important source of natural antioxidants due to its high content of phenolic compounds. The aim of the current study is to maximize the extraction yields of the main phenolic compounds present in olive pomace using innovative green technologies. For this purpose, the present work is divided into two parts. The first part is based on a solubility study of targeted phenolic compounds in various ethanol/water ratios at two different temperatures (20 °C and 50 °C). A computational prediction using COSMO-RS software was applied for the calculation of eventual solubility, which was subsequently confirmed by practical experiments. The determination of the optimal extraction conditions of solvent ratio (EtOH/H2O) (60:40 v/v) and temperature (50 °C) led to the second part of the work, which concerns the intensification of extraction yields. Furthermore, various green extractions using innovative technologies, including accelerated solvent extraction (ASE), ultrasound with its both system (probe (UAE-P) and bath (UAE-B)), bead milling (BM) and microwave (MAE), were carried out and then compared to conventional maceration (CM). Results showed that ASE was the most effective method for extracting phenolic compounds from dried olive pomace powder (5.3 milligrams of tyrosol equivalent (TE) per gram of dried olive pomace powder (DOP)) compared to CM (3.8 mg TE/g DOP).

Development of novel GnRH and Tat48-60 based luminescent probes with enhanced cellular uptake and bioimaging profile.

There is a clear need to develop photostable chromophores for bioimaging with respect to the classically utilized green fluorescent dye fluorescein. Along these lines, we utilized a phosphorescent carboxy-substituted ruthenium(ii) polypyridyl [Ru(bipy)2(mcb)]2+ (bipy = 2,2'-bipyridyl and mcb = 4-carboxy-4'-methyl-2,2'-bipyridyl) complex. We developed two luminescent peptide conjugates of the cell-penetrating peptide Tat48-60 consisting of either [Ru(bipy)2(mcb)]2+ or 5(6)-carboxyfluorescein (5(6)-FAM) tethered on the Lys50 of the peptide through amide bond. We confirmed the efficient cellular uptake of both bioconjugates in HeLa cells by confocal microscopy and flow cytometry and proved that the ruthenium-based chromophore possesses enhanced photostability compared to a 5(6)-FAM-based peptide, after continuous laser scanning. Furthermore, we designed and developed a luminescent agent with high photostability, based on the ruthenium core, that could be selectively localized in cancer cells overexpressing the GnRH receptor (GnRH-R). To achieve this, we took advantage of the tumor-homing character of d-Lys6-GnRH which selectively recognizes the GnRH-R. The [Ru(bipy)2(mcb)]2+-d-Lys6-GnRH peptide conjugate was synthesized, and its cellular uptake was evaluated through flow cytometric analysis and live-cell imaging in HeLa and T24 bladder cancer cells as negative and positive controls of GnRH-R, respectively. Besides the selective targeting that the specific conjugate could offer, we also recorded high internalization levels in T24 bladder cancer cells. The ruthenium(ii) polypyridyl peptide-based conjugates we developed is an intriguing approach that offers targeted cell imaging in the Near Infrared region, and simultaneously paves the way for further advancements in the dynamic studies on cellular imaging.

The impact of lip-split mandibulotomy on patients treated for pT2 oral tongue squamous cell carcinoma: a study of 224 patients.

BACKGROUND: Head and neck surgeons often face a challenge in order to achieve adequate three-dimensional resection of tumours in the oral cavity, especially in the dentate patient. METHODS: We compared the outcomes of lip-split mandibulotomy and trans-oral access, respectively, in patients treated for primary pT2 oral tongue SCC with regard to the status of the resection margins and the incidence of tumour recurrence. RESULTS: Multivariate analysis showed a non-significant effect of the surgical technique used to the reported recurrence, F(1, 224) = 0.350, p = .555 and a significant effect on the margins achieved F(1, 224) = 11.381, p = .001. CONCLUSIONS: Defects after excision of larger and more posterior tumours that are going to be reconstructed with free flaps represent a more probable indication for using an osteotomy access technique. Lip-split mandibulotomy is a low-morbidity technique which can deliver a sound oncological outcome and can be relatively easily taught to less experienced surgeons.

Olive Oil Quality and Authenticity Assessment Aspects Employing FIA-MRMS and LC-Orbitrap MS Metabolomic Approaches.

Edible vegetable oils comprise integral components of humans' daily diet during the lifetime. Therefore, they constitute a central part of dietary-exposome, which among other factors regulates human health. In particular, the regular consumption of olive oil (OO) has been largely accepted as a healthy dietary pattern. Responsible for its recognition as a superior edible oil is its exceptional aroma and flavor. Its unique composition is characterized by high levels of monounsaturated fatty acids and the presence of minor constituents with important biological properties, such as the so-called OO polyphenols. Being a high added value product, OO suffers from extensive fraud and adulteration phenomena. However, its great chemical complexity, variability, and the plethora of parameters affecting OO composition hamper significantly the selection of the absolute criteria defining quality and authenticity, and a reliable and robust methodology is still unavailable. In the current study, Flow Injection Analysis-Magnetic Resonance Mass Spectrometry (FIA-MRMS) was investigated under a metabolic profiling concept for the analysis of Greek Extra Virgin Olive Oils (EVOO). More than 200 monovarietal (Koroneiki) EVOO samples were collected from the main Greek OO producing regions and investigated. Both intact oil and the corresponding polyphenols were analyzed in fast analysis time of 2 and 8 min, respectively. In parallel, an LC-Orbitrap MS platform was used to verify the efficiency of the method as well as a tool to increase the identification confidence of the proposed markers. Based on the results, with FIA-MRMS, comparable and improved projection and prediction models were generated in comparison to those of the more established LC-MS methodology. With FIA-MRMS more statistically significant compounds and chemical classes were identified as quality and authenticity markers, associated with specific parameters, i.e. geographical region, cultivation practice, and production procedure. Furthermore, it was possible to monitor both lipophilic and hydrophilic compounds with a single analysis. To our knowledge, this approach is among the few studies in which two FT-MS platforms combining LC and FIA methods were integrated to provide solutions to quality control aspects of OO. Moreover, both lipophilic and hydrophilic components are analyzed together, providing a holistic quality control workflow for OO.

Olive oil with high polyphenolic content induces both beneficial and harmful alterations on rat redox status depending on the tissue.

Olive oil (OO) possesses a predominant role in the diet of Mediterranean countries. According to a health claim approved by the European Food Safety Authority, OO protects against oxidative stress­induced lipid peroxidation in human blood, when it contains at least 5 mg of hydroxytyrosol and its derivatives per 20 g. However, studies regarding the effects of a total OO biophenols on redox status in vivo are scarce and either observational and do not provide a holistic picture of their action in tissues. Following a series of in vitro screening tests an OO containing biophenols at 800 mg/kg of OO was administered for 14 days to male Wistar rats at a dose corresponding to 20 g OO/per day to humans. Our results showed that OO reinforced the antioxidant profile of blood, brain, muscle and small intestine, it induced oxidative stress in spleen, pancreas, liver and heart, whereas no distinct effects were observed in lung, colon and kidney. The seemingly negative effects of OO follow the recently formulated idea in toxicology, namely the real life exposure scenario. This study reports that OO, although considered a nutritional source rich in antioxidants, it exerts a tissues specific action when administered in vivo.

Reconstructing the Nasal Tip After a Human Bite: A Challenge for the Reconstructive Surgeon.

BACKGROUND: Human facial bites are uncommon and usually cause a wound that presents major challenges for the reconstructive surgeon. PURPOSE: The purpose of this study is to present the advantages and disadvantages of using a superiorly based interpolated nasolabial flap for reconstruction of a nasal tip defect. METHODS: We present a case of serious injury of the nasal tip as a result of a human bite. Early maxillofacial surgical intervention resulted in restoration of the facial anatomy and function of the structures involved. RESULTS AND CONCLUSION: We present a serious mutilating injury of the tip of the nose in a young patient as a result of a human bite. These injuries can be rather destructive and cause a facial defect which may have a significant adverse psychological impact on the patient. Because of the proximity and the skin colour match of the melolabial fold to the nose, nasolabial flaps are a very attractive and reliable reconstructive choice for nasal tip defects. The relative advantages over other reconstructive options and the limitations of an interpolated superiorly based nasolabial flap are discussed. The elaborated use of a nasolabial peninsular pedicled skin flap may deliver a very satisfactory outcome from a functional and cosmetic point of view.

The Polyphenolic Composition of Extracts Derived from Different Greek Extra Virgin Olive Oils Is Correlated with Their Antioxidant Potency.

Olive oil possesses a predominant role in the diet of countries around the Mediterranean basin, whereas it is a known constituent of several sectors of human culture. The polyphenolic composition of olive oil seems to be a key factor in its beneficial biological properties. Based on the above, the aim of this study was to correlate the polyphenolic composition of five extracts derived from a Greek olive oil variety with their antioxidant potency and antimutagenic activities in vitro with chemical-based techniques and cell culture-based assays. According to the results obtained, the polyphenol samples with higher concentration of hydroxytyrosol (HT) were more potent in antioxidant and antimutagenic activity in vitro, as indicated by their ability to scavenge ABTS·+ radical and to protect the strand of plasmid DNA from free radical-induced breaking compared to the corresponding samples with higher levels of tyrosol (T) and its derivatives. However, this observation was not evident in the cell culture model (i.e., the HeLa cervical cancer cell line) to which the tested extracts were administered. Specifically, the T-rich extracts more effectively increased endogenous GSH levels measured by flow cytometry than did the HT-rich compounds. Also, olive oil compounds contributed variously to the expression of genes implicated in the cell antioxidant machinery, as indicated by quantitative PCR. Therefore, the relationship between structure and function in redox regulation is complex and merits the combination of tests. Given that factors like the production and storage regimen of the plants are major determinants of the composition of the generated extracts, we propose that specific conditions should be adopted in order to achieve their maximum biological activity. These results followed by others in the same direction could provide a solid basis for the production of functional foods enriched in olive oil extracts with potential antioxidant action in vivo.

Comparison survey of EVOO polyphenols and exploration of healthy aging-promoting properties of oleocanthal and oleacein.

Olive oil is widely accepted as a superior edible oil. Great attention has been given lately to olive oil polyphenols which are linked to significant health beneficial effects. Towards a survey of Greek olive oil focusing on polyphenols, representative extra virgin olive oils (EVOOs) from the main producing areas of the country and the same harvesting period have been collected and analyzed. Significant differences and interesting correlations have been identified connecting certain polyphenols namely hydroxytyrosol, tyrosol, oleacein and oleocanthal with specific parameters e.g. geographical origin, production procedure and cultivation practice. Selected EVOOs polyphenol extracts, with different oleacein and oleocanthal levels, as well as isolated oleacein and oleocanthal were bio-evaluated in mammalian cells and as a dietary supplement in the Drosophila in vivo model. We found that oleocanthal and oleacein activated healthy aging-promoting cytoprotective pathways and suppressed oxidative stress in both mammalian cells and in flies.

Antioxidant effects of an olive oil total polyphenolic fraction from a Greek Olea europaea variety in different cell cultures.

BACKGROUND: Numerous studies have been carried out concerning the advantageous health effects, especially the antioxidant effects, of olive oil's (OO) individual biophenolic compounds, but none until now for its total phenolic fraction (TPF). Plenty of evidence, in research about nutrition and healthiness, points out that it is the complex mixture of nutritional polyphenols, more than each compound separate, which can synergistically act towards a health result. PURPOSE: The aim of the present study was to examine the antioxidant properties of an extra virgin olive oil (EVOO) total polyphenolic fraction, from a Greek endemic variety of Olea europaea in cell lines. METHODS: EVOO from a Greek endemic variety was used for the extraction of a total polyphenolic fraction, using a green CPE­based method. The redox status [in terms of ROS, GSH, TBARS, protein carbonyls] was assessed at a cellular level, particularly in EA.hy926 endothelial, HeLa, HepG2 hepatic cells and C2C12 myoblasts. Moreover, the levels of glutamate-cysteine ligase catalytic subunit (γ-GCLc) of GSH, one of the most important antioxidant enzymes, were assessed by western blot. RESULTS: According to the results, TPF improves the redox profile of all cell lines, mainly by increasing GSH and its catalytic subunit, while at low, not cytotoxic TPF concentrations there was a decrease in TBARS and carbonyls. Regarding ROS levels a reduction was observed only in the HepG2 cell line, contrary to the other cell lines, that there is no statistically significant difference. CONCLUSION: The TPF appeared to protect cells from oxidative stress due to the strong antioxidant activity of its polyphenols. This could have interesting implications in development of new products based on this olive oil to provide protection and treatment against harmful effects of free radicals.

Evaluation of total phenolic fraction derived from extra virgin olive oil for its antileishmanial activity.

BACKGROUND: Leishmaniasis is a neglected and emerging disease with varying clinical manifestations. The current treatment options rely on limited chemotherapy with serious drawbacks. Thus, there is an increasing interest in the identification of new candidates for designing potent, less toxic and low-cost drugs. PURPOSE: The purpose of this study was to evaluate the potential antileishmanial activity of the total phenolic fraction (TPF) derived from extra virgin olive oil (EVOO) when added in in vitro and in vivo experimental models of Leishmania infection. STUDY DESIGN: We investigated the in vitro antileishmanial activity of TPF against two Leishmania species: a viscerotropic (L. infantum) and a dermotropic (L. major) strain. The antileishmanial effect was also tested in vivo in a murine cutaneous leishmaniasis model using L. major-infected BALB/c mice. METHODS: Separation and analytical methodologies were applied in order to extract the olive oil phenols (TPF) and determine the concentration of the major ones, respectively. The in vitro antileishmanial activity of TPF against promastigotes and intracellular amastigotes was determined by the resazurin cell viability assay. The TPF-induced nitric oxide synthesis by L. infantum and L. major -infected J774A.1 macrophages was determined using the Griess reaction, while the respective generation of reactive oxygen species was assessed by flow cytometry. Moreover, L. major-infected BALB/c mice were treated with TPF and its in vivo therapeutic effect was determined as reduction of the footpad swelling. RESULTS: Our data showed that TPF exhibits inhibitory effect against cell free promastigotes and intracellular amastigotes of both L. infantum and L. major parasite strains. TPF demonstrated to be selectively active against Leishmania amastigotes and its antileishmanial activity was possibly mediated by reactive nitrogen and oxygen intermediates generated from the infected J774A.1 macrophages. Furthermore, administration of TPF in BALB/c mice infected with L. major caused significant reduction of footpad swelling demonstrating in vivo its antileishmanial effect. Based on HPLC-DAD analysis the major components of TPF are tyrosol, hydroxytyrosol, oleacein and oleocanthal. CONCLUSION: This study brings a new low-cost candidate to the leishmaniasis drug discovery pipeline, upon further pharmacological investigation.