A multilevel LC-HRMS and NMR correlation workflow towards foodomics advancement: Application in table olives.

Foodomics employs advanced analytical techniques to provide answers regarding food composition, authenticity control, marker identification and issues related to food quality and safety. Nuclear magnetic resonance (NMR) spectroscopy and chromatography hyphenated to mass spectrometry (MS) are the main analytical platforms used in this field. Nevertheless, they are rarely employed in an integrated manner, and even then, the contribution of each technique remains vague. Table olives (Olea europaea L.) are a food commodity of high economic and nutritional value with an increasing production tendency over the last two decades, which, however, suffers from extensive fraud incidents and quality determination uncertainties. Thus, the current attempt aims towards two axes with the first being the multilevel integration of LC-HRMS and NMR data of the same samples and table olives being the selected matrix. In more detail, UPLC-HRMS/MS-based analysis was compared at different stages within an untargeted metabolomics workflow with an NMR-based study and the complementarity of the two platforms was evaluated. Furthermore, statistical heterospectroscopy (SHY), rarely employed in foodomics, combining the spectroscopic with spectrometric datasets and aiming to increase the confidence level of annotated biomarkers was applied. Amongst these lines, the second parallel axis of this study was the detailed characterization of table olives' metabolome in search for quality markers considering the impact of geographical (from Northern to Southern Greece) and botanical origin (Kalamon, Konservolia, Chalkidikis cultivars), as well as processing parameters (Spanish, Greek). To that end, using deep dereplication tools including statistical methods, with SHY employed for the first time in table olives, different biomarkers, belonging to the classes of phenyl alcohols, phenylpropanoids, flavonoids, secoiridoids and triterpenoids were identified as responsible for the observed classifications. The current binary pipeline, focusing on biomarkers' identification confidence, could be suggested as a meaningful workflow not only in olive-based products, but also in food quality control and foodomics in general.

Clinical Impact of C-myc Oncogenic Diversity on Solid and Lymphoid Malignancies.

INTRODUCTION: Onset and progression of malignant tumors is a multistep process including a variety of gross chromosomal and specific genes' deregulation. Among oncogenes that are frequently altered in solid and also in hematological malignancies, the C-myc (gene locus: 8q24.21) plays a pivotal role. C-myc is a proto-oncogene encoding for a nuclear phosphoprotein implicated in cell cycle progression, apoptosis and cellular differentiation and transformation. OBJECTIVE: The purpose of the current molecular review was to explore the differences of C-myc oncogenic activity in solid and lymphoid malignancies that modify its clinical impact on them. MATERIAL AND METHOD: A systematic review of the literature in the international database PubMed was carried out. The year 2010 was set as a prominent time limit for the publication date of articles in the majority of them, whereas specific references of great importance and historical value in the field of C-myc gene discovery and analysis were also included. The following keywords were used: C-myc, oncogene, signaling pathway, malignancies, carcinoma, lymphoma. A pool of 43 important articles were selected for the present study at the basis of combining molecular knowledge with new targeted therapeutic strategies. RESULTS: C-myc oncogene demonstrates two different mechanisms of deregulation: amplification, mutation and translocation patterns. These particular aspects of gene alteration are unique for solid and non-solid (hematological) malignancies, respectively. CONCLUSIONS: C-myc is characterized by diversity regarding its deregulation mechanisms in malignancies derived from different tissues. C-myc translocation is sporadically combined with amplification ("complicon" formation) or mutations creating exotic genetic signatures. This "bi-phasic" C-myc deregulation model in the corresponding malignant tumor categories clinically affects the corresponding patients, also modifying the targeted therapeutic strategies on them.

C-Jun Transcription Factor Oncogenic Activation in Oral Carcinoma.

INTRODUCTION: Oral carcinogenetic is based on a variety of genomic imbalances (gross chromosome or specific gene alterations) that drive the normal oral mucosa to its neoplastic/dysplastic epithelial form and finally to a totally malignant tissue transformation. In this multi-step procedure, down-regulation of suppressor genes combined with overactivation of oncogenes are two crucial and partially early genetic events involved in the onset and progression of neoplastic/malignant epithelia transformation. More specifically, deregulation of strong transcription factors negatively affects the normal expression of a broad spectrum of genes that are involved in cell proliferation and signalling transduction to the nucleus. OBJECTIVE: The purpose of the current molecular review was to explore the c-Jun (chromosome location: 1p32-p31) transcription factor transformation mechanisms to oncogene in oral squamous cell carcinoma (OSCC). MATERIAL AND METHOD: A systematic review of the literature was carried out by searching in PubMed international database. The year 2010 was set as a prominent time limit for the publication date of the articles in the majority of them, whereas specific references of great importance and historical value in the field of the c-Jun gene discovery and analysis were also included. The following keywords were used: c-Jun, oncogene, signaling pathway, oral, carcinoma, transcription. A pool of 45 important articles were selected for the present study at the basis of combining molecular knowledge with new targeted therapeutic strategies. RESULTS: C-Jun - as a part of the c-Jun/c-Fos transcription factors' complex -critically regulates the expression levels in a variety of genes inside the cellular microenvironment. A broad spectrum of malignancies, including OSCC, demonstrate c-Jun alterations driving the gene to its oncogenic phenotype. Interestingly, c-Jun oncogenic activation is mediated by high-risk human papilloma virus (HR-HPV) persistent infection in significant subsets of these malignancies. CONCLUSIONS: C-Jun was the first oncogene - acting as a strong transcription factor - that was discovered and cloned 35 years ago. C-Jun is the living history of oncogenes and its discovery marks a significant step in the evolution of molecular biology.

Comparative study of Spanish-style and natural cv. Chalkidiki green olives throughout industrial-scale spontaneous fermentation and 12-month storage: safety, nutritional and quality aspects.

Table olives are among the most popular fermented foods and cv. Chalkidiki green table olives are particularly popular in both Greek and international markets. This work aimed at comparatively investigating the effect of processing method on the production of Spanish-style and natural cv. Chalkidiki green olives during fermentation and 12-month storage in brines with different chloride salts composition (NaCl, KCl, CaCl2) at industrial scale. All delivered products were safe with satisfactory color and texture characteristics. Employment of UPLC-HRMS revealed differences in metabolites' profile of polar extracts from olives and brines between the processing methods. Τhe application of alkali treatment drastically decreased the content of hydroxytyrosol and tyrosol in drupes, still an essential amount (1037-2012 and 385-885 mg/kg dry flesh, respectively) of these health-promoting phenolic compounds was retained in all products, even after storage. Noteworthy, fermentation of natural olives in brine (5 % NaCl) yielded in products with significantly lower Na levels in olive flesh (1.7 g/100 g), followed by Spanish-style olives fermented in low (4 %) and high (8 %) NaCl brines (2.7 and 5.2 g Na/100 g, respectively), supporting the efforts toward the establishment of table olives as functional food. Moreover, maslinic and oleanolic acids content was 1.5-2-fold higher in the natural table olives compared to the Spanish-style ones owing to the detrimental effect of alkali treatments. The processing method did not exert a differential effect on α-tocopherol content of olives. Sensory analysis indicated that all the final products were acceptable by consumers, with a slight preference for Spanish-style green olives fermented in brines with 50 % lower NaCl content. Present findings could be beneficial to the ongoing endeavor directed for the establishment of table olives as a source of bioactive compounds that concerns both industrial and scientific communities.

Management of the atrophic mandibular fractures: a ten-year prospective study of 48 injuries.

PURPOSE: The purpose of this study was to (a) record and evaluate the epidemiological data relevant to the fractures of the atrophic mandible in the Greek population (b) present our experience in the management of these difficult injuries and (c) compare our results to the outcomes of other similar studies and discuss the postoperative complications. METHODS: A prospective analysis of all the edentulous patients with fractures of the atrophic mandible treated at the Oral and Maxillofacial Surgery Department of K.A.T General Hospital of Athens in Greece was performed from November 2012 to December 2022. Age, gender and medical history of the patient, etiology and site of the fracture, classification of atrophy, type of surgical approach, type of osteosynthesis and postoperative complication. RESULTS: 34 patients were included in the present study and 48 fractures of the atrophic edentulous mandible were managed surgically. 22 fractures were classified as class II of atrophy, 21 fractures as class III and 5 injuries as class I. In 32 patients we used an extraoral approach and only 2 patients were treated with an intraoral access. 44 fractures were treated with a 2.0 mm locking reconstruction plate and only 4 injuries of class I atrophy were treated with mini plates. CONCLUSIONS: Clinical practice has confirmed that for these cases an extraoral approach followed by stable fixation with a 2.0 mm reconstruction locking plate can deliver excellent results. Our findings show that the routine use of primary bone grafts is not necessary and can be reserved for more complex cases.

Epidemiology, etiopathogenesis, and management of MRONJ: A European multicenter study.

INTRODUCTION: The purpose of this European multicenter study was to describe the general characteristics and risk factors of MRONJ lesions as well as their clinical diagnosis and management at different European Oral and Maxillofacial Surgery centers, in order to minimize selections biases and provide information about the epidemiology, etiopathogenesis, and the current trends in the treatment of MRONJ across Europe. MATERIALS AND METHODS: The following data were registered for each patient: gender; age at MRONJ diagnosis; past medical history; indication for antiresorptive or antiangiogenic therapy; type of antiresorptive medication; local risk factor for MRONJ; MRONJ Stage; anatomic location and symptoms; treatment; surgical complications; recurrence. RESULTS: A total of 537 patients (375 females, 162 males) with MRONJ were included. Statistically significant associations were found between patients with metastatic bone disease and recurrences (P < 0.0005) and between advanced MRONJ stages (stages 2 and 3) and recurrences (P < 0.005). Statistically significant associations were also found between male gender and recurrences (P < 0.05), and between MRONJ maxillary sites and recurrences (P < 0.0000005). CONCLUSIONS: A longer mean duration of antiresorptive medications before MRONJ onset was observed in patients affected by osteoporosis, whereas a shorter mean duration was observed in all metastatic bone cancer patients, and in particular in those affected by prostate cancer with bone metastases or multiple myeloma. Surgery plays an important role for the management of MRONJ lesions.

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.