Chemical characteristics and biological activity screening of Pistacia lentiscus mastic gum and leaves from Türkiye.

BACKGROUND: Mastic gum is a resin that is produced by Pistacia lentiscus. It has many traditional uses, dating from ancient times, such as the treatment of gastrointestinal disorders and as a food additive. In this study, the leaves and mastic gum of trees of different ages from Karaburun and the Cesme peninsula in Türkiye were examined chemically and biologically. Flavonoids, and phenolic and fatty acid components were evaluated by a liquid chromatography system coupled with high resolution mass spectrometry (LC-HRMS) and gas chromatography with flame ionization detection (GC-FID). Cytotoxicity was screened against several cancer and healthy cell lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Inducible nitric oxide synthase (iNOS) inhibition was determined on lipopolysaccharide (LPS)-induced murine macrophage cell line (RAW 264.7) cells. Antiviral activity was measured against avian coronavirus using an in ovo virucidal antiviral activity assay. RESULTS: The main phenolic constituents of the gum were found to be salicylic, rosmarinic, and caffeic acids whereas the most abundant compounds detected were flavonoids in the leaf extracts. The most abundant fatty acids in hexane extracts were palmitic and oleic acids. All gum extracts except 3-year-old gum had significant cytotoxic activity on HeLa (IC50 1.74 ± 0.03-4.76 ± 0.95) and PC-3 (0.64 ± 0.25-6.22 ± 1.40) cells. Moreover, reducing virus activity by fivefold or sixfold logarithmically between the range of 5-10 µg g-1 of 30-year-old gum extracts underscored the biological activity. CONCLUSION: In ovo antiviral activity studies on the P. lentiscus were conducted for the first time. The mastic gum and leaves obtained from P. lentiscus may have strong potential in terms of their chemical content and antiviral and cytotoxic activity. As a consequence of these properties, it is a sustainable, renewable natural resource that can be used as an additive and flavoring in the food and pharmaceutical industries. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Antibacterial study of carbopol-mastic gum/silver nanoparticle-based topical gels with carvacrol/neem bark extract in vitro.

BACKGROUND: Resistance to antimicrobial drugs as a result of prolonged use usually results in clinical failure, especially in wound infections. Development of effective antimicrobial therapeutics for the management of infected wounds from a natural source with improved therapeutic effects is a pressing need. OBJECTIVE: In this study, carbopol-mastic gum-based topical gels were loaded with silver nanoparticles in combination with either neem bark extract or carvacrol oil. The effect of combining silver nanoparticles with neem bark extract or the essential oil carvacrol in the prepared gel formulations was investigated on selected bacterial strains. METHOD: The prepared gels were characterised by Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM) and ultraviolet-visible (UV-vis) spectroscopy, followed by antimicrobial analysis against selected strains of bacteria. RESULTS: There was no interaction between the loaded natural extract or essential oil and the polymer used for the preparation of the formulations, which was visible from the FTIR spectra of the formulations. The gels were selective and effective against selected strains of bacteria. However, the combination of the silver nanoparticles with essential oil or natural extract in some of the gel formulations rendered the formulation ineffective against some of the bacterial strains. CONCLUSION: The gel formulations were effective against bacterial strains such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis which are the common causes of wound infections. Incorporation of silver nanoparticles into the topical formulations with natural extracts is usually a good approach to overcome antibiotic-resistant infections. However, the combination of antibacterial agents must be managed carefully.

Phytochemical investigation of Pistacia lentiscus L. var. Chia leaves: A byproduct with antimicrobial potential.

Pistacia lentiscus L. var. Chia belongs to the Anacardiaceae family, and it is cultivated only in the south part of Chios island, in Greece. Even though it is renowned for its unique resin, Chios mastic gum (CMG), the tree leaves have also been used in traditional medicine, while the annual pruning generates a large biomass of unused by-products. Thus, the aim of the present study was the detailed phytochemical investigation of P. lentiscus var. Chia leaves towards the search of antimicrobial agents. UPLC-HRMS & HRMS/MS based dereplication methods led to the detailed characterization of the aqueous leaf extract. In addition, twelve compounds were isolated and purified from the methanol extract and were identified using spectroscopic and spectrometric methods (NMR, HRMS) belonging to phenolic acids, tannins, flavonoids and terpenes, with the most interesting being 2-hydroxy-1,8-cineole ß-D-glucopyranoside which was isolated for the first time in the Anacardiaceae family. Remarkably, based on NMR data, methanol and aqueous extracts were found to be particularly rich in shikimic acid, a valuable building block for the pharmaceutical industry, for instance in the synthesis of the active ingredient of Tamiflu®, oseltamivir. Finally, extracts (EtOAc, MeOH, H2O) and major compounds i.e., shikimic acid, 2-hydroxy-1,8-cineole ß-D-glucopyranoside and myricitrin were evaluated for their antimicrobial properties. MeOH and H2O mastic leaf extracts as well as myricitrin and, particularly, 2-hydroxy-1,8-cineole ß-D-glucopyranoside showed significant selective activity against pathogenic Mucorales, but not against Aspergilli (Aspergillus nidulans, Aspergillus fumigatus), Candida albicans or bacteria (Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis).

Mastic (Pistacia lentiscus) gum and oral health: a state-of-the-art review of the literature.

The aim of this state-of-the-art review is to provide a comprehensive overview of the various therapeutic effects of Mastic (Pistacia lentiscus) gum on oral health. The search of the literature was conducted across thirteen databases for relevant publications published through May 2022 in English, Arabic, or Greek using a combination of keywords and phrases. Out of 246 papers, the search procedure identified 14 papers for inclusion. Mastic gum displayed antibacterial and antimicrobial properties and inhibited plaque accumulation, constituting a beneficial adjuvant in caries prevention. In the treatment and prevention of periodontal diseases, Pistacia lentiscus essential oil provided effective antibacterial activity against a variety of periodontal bacteria as well as anti-inflammatory properties. For oral cancer, several clinical trials revealed interesting results against cell proliferation, induction of apoptosis, and regulation of intracellular signaling pathways. This indicates the potential of Mastic gum to serve as a preventive and therapeutic agent for oral mucosa inflammation and oral cancer. No notable toxic or side effects were reported in the clinical trials reviewed. This review highlights the various beneficial effects of Mastic gum in the prevention and potential treatment of oral diseases. Further research targeting Pistacia lentiscus products is required in order to validate and utilize these products to prevent and to treat oral health diseases.

The Effect of Mastic Chios Supplementation in Inflammatory Bowel Disease: A Systematic Literature Review.

Mastic Chios is a natural treasure of Greece that offers many potential benefits in various disorders due its antioxidant and anti-inflammatory properties. This systematic review aimed to investigate the effect of Mastic Chios supplementation in inflammatory bowel disease (IBD). A systematic search was conducted following PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines across PubMed/Medline and EBSCO databases until February 2023. Clinical studies that examined individuals of any age being diagnosed with IBD as well as preclinical studies using animals with experimental colitis were considered eligible for inclusion. Studies included were assessed for methodological quality. Eight articles were eligible for inclusion, with seven conducted in Greece and one study in Iran. Most of the studies were randomized controlled trials (RCTs) and one was a pilot study. The intervention period varied between 3 and 7 days for animal studies and 4 weeks to 6 months for human studies. Mastic Chios supplementation varied between 2.2 and 2.8 g/day for clinical studies. The majority of studies concluded that Mastic Chios has significant antioxidant and inflammatory effects in IBD. This systematic review suggests that Mastic Chios may have a positive effect on the regulation and management of IBD. However, more high-quality clinical trials are needed to provide reliable and concrete conclusions about the effectiveness of Mastic Chios in this population. Such studies will further support the use of Mastic Chios as a reliable therapeutic option for IBD.

Masticadienonic acid from Chios mastic gum mitigates colitis in mice via modulating inflammatory response, gut barrier integrity and microbiota.

BACKGROUND: Chios mastic gum (CMG) is a traditional Greek medicine used to treat a variety of gastrointestinal disorders, including inflammatory bowel disease (IBD). However, the bioactive compounds of CMG and the mechanisms of action for controlling of IBD remain unknown. PURPOSE: Masticadienonic acid (MDA) is one of the most abundant constituents isolated from CMG. This study aims to investigate the potential effects and underlying mechanisms of MDA in the pathogenesis of colitis. METHODS: The effects of MDA were evaluated using a dextran sulphate sodium (DSS)-induced acute colitis mouse model. The body and spleen weight and colon length and weight were measured and the clinical symptoms were analysed. Blood samples were collected to analyse the level of serum inflammatory markers. Colon tissues were processed for histopathological examination, evaluation of the epithelial barrier function, and investigation of the probable mechanisms of action. The gut microbiota composition was also studied to determine the mechanism for the beneficial effects of MDA on IBD. RESULTS: MDA could ameliorate the severity of IBD by increasing the body weight and colon length, reducing spleen weight, disease activity index, and histological score. MDA treatments reduce the release of serum inflammatory cytokines tumour necrosis factor-alpha (TNFα), interleukin 1 beta (IL-1ß), and interleukin 6 (IL-6) via inhibiting the MAPK and NF-κB signalling pathways. MDA supplementation could also improve the intestinal barrier function by activating the NF-E2-related factor-2 (Nrf2) signalling pathway and restoring the expression of tight junction proteins zonula occludens-1 (ZO-1) and occludin. In addition, MDA administration modulates the gut microbiota composition in DSS-induced colitis mice. CONCLUSION: The results indicate that MDA attenuated experimental colitis by restoring intestinal barrier integrity, reducing inflammation, and modulating the gut microbiota. The present study provides novel insights into CMG-mediated remission of IBD and may facilitate the development of preventive and therapeutic strategies for IBD.

Nutrigenetic Interactions Might Modulate the Antioxidant and Anti-Inflammatory Status in Mastiha-Supplemented Patients With NAFLD.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease with no therapeutic consensus. Oxidation and inflammation are hallmarks in the progression of this complex disease, which also involves interactions between the genetic background and the environment. Mastiha is a natural nutritional supplement known to possess antioxidant and anti-inflammatory properties. This study investigated how a 6-month Mastiha supplementation (2.1 g/day) could impact the antioxidant and inflammatory status of patients with NAFLD, and whether genetic variants significantly mediate these effects. We recruited 98 patients with obesity (BMI ≥ 30 kg/m2) and NAFLD and randomly allocated them to either the Mastiha or the placebo group for 6 months. The anti-oxidative and inflammatory status was assessed at baseline and post-treatment. Genome-wide genetic data was also obtained from all participants, to investigate gene-by-Mastiha interactions. NAFLD patients with severe obesity (BMI > 35kg/m2) taking the Mastiha had significantly higher total antioxidant status (TAS) compared to the corresponding placebo group (P value=0.008). We did not observe any other significant change in the investigated biomarkers as a result of Mastiha supplementation alone. We identified several novel gene-by-Mastiha interaction associations with levels of cytokines and antioxidant biomarkers. Some of the identified genetic loci are implicated in the pathological pathways of NAFLD, including the lanosterol synthase gene (LSS) associated with glutathione peroxidase activity (Gpx) levels, the mitochondrial pyruvate carrier-1 gene (MPC1) and the sphingolipid transporter-1 gene (SPNS1) associated with hemoglobin levels, the transforming growth factor-beta-induced gene (TGFBI) and the micro-RNA 129-1 (MIR129-1) associated with IL-6 and the granzyme B gene (GZMB) associated with IL-10 levels. Within the MAST4HEALTH randomized clinical trial (NCT03135873, www.clinicaltrials.gov) Mastiha supplementation improved the TAS levels among NAFLD patients with severe obesity. We identified several novel genome-wide significant nutrigenetic interactions, influencing the antioxidant and inflammatory status in NAFLD. Clinical Trial Registration: ClinicalTrials.gov, identifier NCT03135873.

Development, Validation and Application of a UHPLC-MS Method for the Quantification of Chios Mastic Gum Triterpenoids in Human Plasma.

Chios mastic gum is the resinous secretion obtained from the barks of the shrub Pistacia lentiscus var. Chia, which is endemic to the Greek island of Chios. Since antiquity, Chios mastic gum has found several uses as a phytotherapeutic remedy, primarily for the treatment of gastrointestinal disorders while recently, Chios mastic gum was also recognized by EMA as an herbal medicinal product with specific indications. Chios mastic gum's biological properties are attributed to triterpenes which comprise the major chemical group (approx. 70%) and notably isomasticadienonic acid and masticadienonic acid. However, due to their structural characteristics, the isolation thereof in high yield and purity is challenging and since they are not commercially available, pharmacological studies aiming to assess their biological properties are limited. In the present work, mastic's phytochemical investigation by UPLC-HRMS is followed by the isolation and characterization of isomasticadienonic acid and masticadienonic acid to be used as analytical standards for their accurate and reliable quantification in human plasma. A UHPLC-tQ-MS method that was developed and validated (in terms of specificity, linearity, limit of quantification, accuracy and precision), for the direct quantification of the targeted compounds in the low ng/mL range of concentration, was subsequently implemented on plasma samples of healthy volunteers thus demonstrating its fitness for purpose. The results presented herein might provide insight to the understanding of this traditional natural product consumed notably for its anti-inflammatory, antioxidant and lipid lowering properties. Moreover, this method might serve as a starting point for any study aiming to monitor bioactive triterpenes in biological fluids.

Structure and organization of the secretion apparatus of the mastic tree (Pistacia lentiscus L.) and LC-HRMS analysis of leaf extracts.

MAIN CONCLUSION: The quantitative profile of the biochemicals secreted by summer and winter leaves, present noticeable differences and appear to be qualitatively different from the biochemical profile of the commercially valuable mastic. The anatomy of the root and the primary and secondary shoot as well as that of the summer and winter leaves of P. lentiscus was thoroughly investigated. The secreting network was tracked throughout the plant axis, from the root to the leaves, and the active secreting cells of the duct epithelium were localized, while the secondary metabolites produced within the cells of the summer and winter leaf tissues were identified histochemically. Numerous phytochemicals were identified in the leaf extracts with UHPLC-qTOF MS analysis. The analyzed extracts from summer and winter leaves displayed similar qualitative profile, although quantitative differences were evident, since, during the summer, the leaves tend to synthesize the more complex amongst the identified compounds. The phytochemical profile of the leaf extracts turns to be completely different compared to that of the valuable mastic harvested from the injured trunks. Many of the compounds common in mastic were not detected in the analyzed leaves samples. The numerous secreting ducts either fail to form a unified network, so composition of the secreted material varies in the different organs of the plant or they compose a continuous network, but the biochemical profile of the secreted material differs along the plant axis. Such a detailed investigation of the secretion network of the mastic tree may assist the improvement of the yield and promote the production of valuable phytochemicals through in vitro cultures.

Effect of Mastiha supplementation on NAFLD: The MAST4HEALTH Randomised, Controlled Trial.

SCOPE: Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease with poor therapeutic strategies. Mastiha possesses antioxidant/anti-inflammatory and lipid-lowering properties. The authors investigate the effectiveness of Mastiha as a nonpharmacological intervention in NAFLD. METHODS AND RESULTS: Ninety-eight patients with NAFLD in three countries (Greece, Italy, Serbia) are randomly allocated to either Mastiha or Placebo for 6 months, as part of a multicenter, randomized, double-blind, placebo-controlled, parallel-group clinical trial. The authors assess NAFLD severity via magnetic resonance imaging (MRI) scanning and LiverMultiScan technique and evaluate the effectiveness of Mastiha through medical, anthropometric, biochemical, metabolomic, and microbiota assessment. Mastiha is not superior to Placebo on changes in iron-corrected T1 (cT1) and Liver Inflammation Fibrosis score (LIF) in entire patient population; however, after BMI stratification (BMI ≤ 35 kg m-2 and BMI > 35 kg m-2 ), severely obese patients show an improvement in cT1 and LIF in Mastiha versus Placebo. Mastiha increases dissimilarity of gut microbiota, as shown by the Bray-Curtis index, downregulates Flavonifractor, a known inflammatory taxon and decreases Lysophosphatidylcholines-(LysoPC) 18:1, Lysophosphatidylethanolamines-(LysoPE) 18:1, and cholic acid compared to Placebo. CONCLUSION: Mastiha supplementation improves microbiota dysbiosis and lipid metabolite levels in patients with NAFLD, although it reduces parameters of liver inflammation/fibrosis only in severely obese patients.

Contact Dermatitis From Gum Mastic (Pistacia lentiscus) and Gum Storax (Liquidambar styraciflua) in Mastisol-Allergic Patients.

BACKGROUND: Mastisol Liquid Adhesive is widely used on the skin, especially after surgical procedures. It contains gum mastic, gum storax, methyl salicylate, and ethanol. OBJECTIVE: The aims of the study were to review our experience patch testing patients allergic to Mastisol and to assess coreacting substances. METHODS: We identified 18 patients who were allergic to Mastisol. Most of these had a history of postoperative or cardiac electrode dermatitis and underwent patch testing with multiple surgically related substances, including ingredients of Mastisol, compound tincture of benzoin, and fragrance-related ingredients and botanicals. RESULTS AND CONCLUSIONS: Among Mastisol-allergic patients, 13 (72%) of 18 were allergic to gum mastic, whereas 7 (44%) of 16 were allergic to gum storax. There was frequent coreactivity with various fragrance-related materials, including Majantol, Styrax benzoin, Myroxylon balsamum, Myroxylon pereirae, propolis, and others. Two gum mastic-allergic patients had positive patch tests with hydroperoxides of linalool and several other linalool-containing essential oils. As gum mastic contains linalool, it may explain some gum mastic reactions. Among patients without a history of postoperative contact dermatitis, 1 (0.4%) of 250 was patch test positive for gum mastic. This patient had allergic contact dermatitis from fragrances, so the gum mastic reaction was likely a true-positive relevant reaction.

Chemical Characterization and Biological Activity of the Mastic Gum Essential Oils of Pistacia lentiscus Var. Chia from Turkey.

The essential oils (EOs) were isolated by hydrodistillation from wild and cultivated Pistacia lentiscus L. var. chia-mastic gum tree (Anacardiaceae) from two natural habitats, namely from Cesme-Uzunkoy (1) and Mordogan (2), and one cultivated source, Cesme-Germiyan (3), in Izmir, Turkey. This comparative study evaluated the chemical composition and biological activity of mastic gum essential oils (MGEOs). For this purpose, MGEOs 1-3 were analyzed by gas chromatography-flame ionization detection (GC-FID), gas chromatography-mass spectrometry (GC-MS), and chiral GC for α-pinene. Laboratory assays were conducted to assess for potential in vitro cytotoxicity (multiple in vitro cancer cell lines), antimicrobial properties (five bacterial species and yeast), anti-inflammatory activity (inhibition of inducible nitric oxide synthase, iNOS), and the attraction of Ceratitis capitata (Mediterranean fruit fly, medfly), respectively. Chemical analysis indicated that MGEOs 1 and 2 were rich in α-pinene (56.2% and 51.9%), myrcene (20.1% and 18.6%), and ß-pinene (2.7% and 3.1%), respectively; whereas MGEO-3 was characterized by a high level of α-pinene (70.8%), followed by ß-pinene (5.7%) and myrcene (2.5%). Chiral GC analyses showed that concentration ratios between (-)/(+)-α-pinene and (-)-α-pinene/myrcene allowed for differentiation between wild and cultivated MGEO sources. In biological assays, MGEOs 1-3 did not exhibit significant antimicrobial effects against the pathogens evaluated and were not strong attractants of male medflies; however, all three MGEOs displayed a dose-dependent inhibition of iNOS, and MGEOs 1 and 2 exhibited selective in vitro cytotoxicity against human cancer cells. These results suggest that wild-type mastic gum oils from Cesme and Mordogan (MGEOs 1 and 2) are potential sources of beneficial products and warrant further investigation.

Traditional uses, phytochemistry and pharmacology of Chios mastic gum (Pistacia lentiscus var. Chia, Anacardiaceae): A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Chios mastic gum constitutes a unique Greek product, produced exclusively in the southern part of the island of Chios. References about its use from local populations for the treatment of gastrointestinal disorders or as a cosmetic agent can even be encountered in ancient texts of Galen, Theophrastus and Dioscorides. Nowadays, this versatile resin has been rediscovered, not only as a traditional remedy and aromatic agent, but as a potent phytotherapeutic product with various biological properties. AIM OF THE STUDY: The aim of this study is to quote the summation of the ethnopharmacology, phytochemical profile and pharmacological properties of the resin of Pistacia lentiscus var. Chia and thus provide the scientific community with a summary of the research conducted so far. Furthermore, perspectives and uses are being discussed and studied so as to broaden the field of its applications. MATERIALS AND METHODS: A comprehensive review of the literature on Pistacia lentiscus var. Chia was performed using as resources scientific databases such as Scopus, Sciencedirect, Pubmed and Web of science, studies and traditional books provided by the Chios Mastiha Growers Association as well as PhD and Master' s theses. RESULTS: Chios mastic gum has been used as a traditional medicine over the last 2500 years. More than 120 chemical compounds have been identified in the resin and the major components are a natural polymer, acidic and neutral triterpenes and volatile secondary metabolites. Several plant extracts and compounds have been studied for their antibacterial, anti-inflammatory, antioxidant, anti-ulcer, anti-diabetic, cardioprotective and anti-cancer properties in vitro and in vivo. Clinical interventions and trials have also showed the therapeutic potential of Chios mastic gum. In 2015 Pistacia lentiscus L., resin (mastic) was recognized as a herbal medicinal product with traditional use by the European Medicines Agency (EMA) with two therapeutic indications (mild dyspeptic disorders & skin inflammation/healing of minor wounds). Over the last years, Chios mastic gum is widely involved in medicinal products, food supplements and cosmetics and has become object of study, also in the field of Pharmacotechnology. CONCLUSIONS: Chios mastic's beneficial properties have been demonstrated in the treatment of gastrointestinal disorders, wound healing, skin inflammations, plasma lipid and blood sugar reduction and oral care. These properties are attributed to triterpenes and volatile compounds. However, because of the resin's chemical complexity and the lack of commercial standards for its main compounds, there is a notable gap in literature concerning the biological evaluation of CMG's isolated components. Therefore, future research should focus on the development of efficient extraction, isolation and analysis techniques in order to unravel CMG's full pharmacological potential.

Randomized Controlled Phase 2a Study of RPh201 in Previous Nonarteritic Anterior Ischemic Optic Neuropathy.

BACKGROUND: No proven treatment exists for nonarteritic anterior ischemic optic neuropathy (NAION), either in the acute or late phase. OBJECTIVE: To assess safety and changes in visual function and structure after RPh201/placebo treatment in participants with previous NAION. DESIGN AND SETTING: Phase 2a, single-site, prospective, randomized, placebo-controlled, double-masked trial (registration NCT02045212). MAIN OUTCOMES MEASURES: Early Treatment Diabetic Retinopathy Study best-corrected visual acuity (BCVA), visual fields, retinal nerve fiber layer, and visual evoked potential at weeks 13, 26, and after a 13-week wash-out ("off-drug") period; and safety. STUDY POPULATION: Twenty-two participants aged 18 years or older with previous NAION. INTERVENTION(S): RPh201 (20 mg) or placebo (cottonseed oil vehicle) administered subcutaneously twice weekly at the study site. RESULTS: Thirteen men and 9 women were randomized, of which 20 completed all visits. The mean (±SD) age was 61.0 ± 7.6 years. In a post hoc analysis, after 26 weeks of treatment, BCVA improved by ≥15 letters in 4/11 (36.4%) eyes with RPh201, compared to 1/8 (12.5%) eyes with placebo (P = 0.24). Overall, 7/11 (63.6%) of participants on RPh201 showed some improvement in BCVA, compared with 3/8 (37.5%) on placebo (P = 0.26). Improvement in BCVA from a calculated baseline was 14.8 ± 15.8 letters for RPh201 and 6.6 ± 15.3 for placebo (P = 0.27). Of the 154 adverse effects (AEs), 52 were considered related to the study procedures/treatment. Across the study and 1,017 injections, the most frequently reported AE was injection site pain (23 events in 5 participants). There were no clinically significant changes in vital signs or laboratory values. CONCLUSIONS: This Phase 2a was designed to assess safety, feasibility, and explore potential efficacy signals in treating previous NAION with RPh201. No safety concerns were raised. The results support a larger trial in patients with previous NAION.

Ancient DNA from mastics solidifies connection between material culture and genetics of mesolithic hunter-gatherers in Scandinavia.

Human demography research in grounded on the information derived from ancient DNA and archaeology. For example, the study on the early postglacial dual-route colonisation of the Scandinavian Peninsula is largely based on associating genomic data with the early dispersal of lithic technology from the East European Plain. However, a clear connection between material culture and genetics has been lacking. Here, we demonstrate that direct connection by analysing human DNA from chewed birch bark pitch mastics. These samples were discovered at Huseby Klev in western Sweden, a Mesolithic site with eastern lithic technology. We generated genome-wide data for three individuals, and show their affinity to the Scandinavian hunter-gatherers. Our samples date to 9880-9540 calBP, expanding the temporal range and distribution of the early Scandinavian genetic group. We propose that DNA from ancient mastics can be used to study environment and ecology of prehistoric populations.

In vitro and in vivo evaluation of the genotoxic and antigenotoxic potential of the major Chios mastic water constituents.

Chios mastic products are well-known for their broad applications in food industry, cosmetics, and healthcare since the antiquity. Given our recent finding that Chios mastic water (CMW) exerts antigenotoxic action, in the present study, we evaluated the genotoxic as well as the antigenotoxic potential of the four major compounds of CMW, namely, verbenone, α-terpineol, linalool, and trans-pinocarveol. The cytokinesis block micronucleus (CBMN) assay in cultured human lymphocytes and the Drosophila Somatic Mutation And Recombination Test (SMART), also known as the wing spot test, were employed. None of the four major CMW's constituents or their mixtures showed genotoxic or recombinogenic activity in either of the assays used. Co-treatment of each of the constituents with MMC revealed that all except trans-pinocarveol exerted antigenotoxic potential. Moreover, co-administration of verbenone with linalool or α-terpineol presented statistically significant reduction of MMC-induced mutagenicity. In conclusion, the major CMW constituents were shown to be free of genotoxic effects, while some exerted antigenotoxic activity either alone or in combinations, suggesting synergistic phenomena. Our results provide evidence on the key antigenotoxicity effectors of the plant extract CMW.

Human Adenocarcinoma Cell Line Sensitivity to Essential Oil Phytocomplexes from Pistacia Species: a Multivariate Approach.

Principal component analysis (PCA) multivariate analysis was applied to study the cytotoxic activity of essential oils from various species of the Pistacia genus on human tumor cell lines. In particular, the cytotoxic activity of essential oils obtained from P. lentiscus, P. lentiscus var. chia (mastic gum), P. terebinthus, P. vera, and P. integerrima, was screened on three human adenocarcinoma cell lines: MCF-7 (breast), 2008 (ovarian), and LoVo (colon). The results indicate that all the Pistacia phytocomplexes, with the exception of mastic gum oil, induce cytotoxic effects on one or more of the three cell lines. PCA highlighted the presence of different cooperating clusters of bioactive molecules. Cluster variability among species, and even within the same species, could explain some of the differences seen among samples suggesting the presence of both common and species-specific mechanisms. Single molecules from one of the most significant clusters were tested, but only bornyl-acetate presented cytotoxic activity, although at much higher concentrations (IC50 = 138.5 µg/mL) than those present in the essential oils, indicating that understanding of the full biological effect requires a holistic vision of the phytocomplexes with all its constituents.

Dietary mastic oil extracted from Pistacia lentiscus var. chia suppresses tumor growth in experimental colon cancer models.

Plant-derived bioactive compounds attract considerable interest as potential chemopreventive anticancer agents. We analyzed the volatile dietary phytochemicals (terpenes) present in mastic oil extracted from the resin of Pistacia lentiscus var. chia and comparatively investigated their effects on colon carcinoma proliferation, a) in vitro against colon cancer cell lines and b) in vivo on tumor growth in mice following oral administration. Mastic oil inhibited - more effectively than its major constituents- proliferation of colon cancer cells in vitro, attenuated migration and downregulated transcriptional expression of survivin (BIRC5a). When administered orally, mastic oil inhibited the growth of colon carcinoma tumors in mice. A reduced expression of Ki-67 and survivin in tumor tissues accompanied the observed effects. Notably, only mastic oil -which is comprised of 67.7% α-pinene and 18.8% myrcene- induced a statistically significant anti-tumor effect in mice but not α-pinene, myrcene or a combination thereof. Thus, mastic oil, as a combination of terpenes, exerts growth inhibitory effects against colon carcinoma, suggesting a nutraceutical potential in the fight against colon cancer. To our knowledge, this is the first report showing that orally administered mastic oil induces tumor-suppressing effects against experimental colon cancer.

Evaluation of Biological Activity of Mastic Extracts Based on Chemotherapeutic Indices.

BACKGROUND: Most previous mastic investigators have not considered its potent cytotoxicity that may significantly affect the interpretation of obtained data. In the present study, we re-evaluated several biological activities of mastic extracts, based on chemotherapeutic indexes. MATERIALS AND METHODS: Pulverized mastic gum was extracted with n-hexane and then with ethyl acetate or independently with methanol or n-butanol. Tumor specificity (TS) of the extracts was determined by their cytotoxicity against human malignant and non-malignant cells. Antibacterial activity was determined by their cytotoxicity against bacteria and normal oral cells. Antiviral activity was determined by their protection of viral infection and cytotoxic activity. Cytochrome P-450 (CYP) 3A4 activity was measured by ß-hydroxylation of testosterone. RESULTS: Ethyl acetate extract showed slightly higher tumor specificity (TS=2.6) and one order higher antibacterial activity (selectivity index (SI)=0.813) than other extracts (TS=1.4-2.5; SI=0.030-0.063). All extracts showed no anti-human immunodeficiency virus (HIV) activity, but some anti-herpes simplex virus (HSV) activity, which was masked by potent cytotoxicity. They showed strong inhibitory activity against CYP3A4. CONCLUSION: Ethyl acetate extraction following the removal of cytotoxic and CYP3A4 inhibitory substances by n-hexane can enhance antitumor and antibacterial activity of mastic.

Quality profile determination of Chios mastic gum essential oil and detection of adulteration in mastic oil products with the application of chiral and non-chiral GC-MS analysis.

The determination of mastic oil profile, with emphasis on its chiral characteristics, could serve as a method for detecting adulteration in products found in the market with a claim of mastic oil content aiming towards protecting it from counterfeiting. Furthermore the evaluation of the raw material is crucial, as the profile is potentially affected by factors as mastic origin and storage time. Thus 45 authentic mastic oil samples were analyzed by GC-MS employing a chiral column and content limits for all major constituents were determined. The chiral GC-MS analysis proved that selected concentration ratios between these constituents, namely those of (-)/(+)-α-pinene (≤1:100) and (-)-α-pinene/myrcene (1.9:100-11:100) could serve as markers for the determination of mastic oil authenticity. Employing this methodology, the analysis of 25 mastic oils contained in cosmetic and dietary products, as well as an artificial mastic oil sample, exhibited several differentiations that could indicate adulteration either with artificial essential oils or volatile compounds, or the use of aged mastic oil.