Abstract

Background: Gastrointestinal tract cancers represent a significant worldwide health concern, accounting for almost one-third of cancer-related deaths. The existing chemotherapy drugs used in gastrointestinal cancers are ineffective, so prognosis is poor, recurrence and metastasis rates are high, and survival time remains short, necessitating the development of novel antitumor drugs that exhibit low toxicity and less potential for the development of drug resistance. This challenge is considerable, but evidence from the past decades supports the medicinal properties and functionalities of bioactive compounds such as flavonoids and acid phenolics with anticancer activities. Our purpose was to find data on the relationship between gastrointestinal cancer and bioactive compounds from Prunus species, focusing on their molecular mechanisms of action. Results: Studies highlight the potential of bioactive compounds from Prunus species to modulate the cancer cell signaling pathways involved in gastrointestinal tumorigenesis. Conclusions: The studies reviewed suggest that polyphenols from Prunus species exhibit promising gastrointestinal anticancer activities and could represent an adjunctive therapeutic strategy in cancer treatment. Further studies are necessary to validate these compounds' therapeutic potential and their feasibility as cost-effective treatments for cancer.

Abstract

Healthcare-associated infections (HAIs), previously known as nosocomial infections, represent a significant threat to healthcare systems worldwide, prolonging patient hospital stays and the duration of antimicrobial therapy. One of the most serious consequences of HAIs is the increase in the rate of antibiotic resistance (AR) generated by the prolonged, frequent, and sometimes incorrect use of antibiotics, which leads to the selection of resistant bacteria, making treatment difficult and expensive, with direct consequences for the safety of patients and healthcare personnel. Therefore, timely and accurate diagnosis of HAIs is mandatory to develop appropriate infection prevention and control practices (IPC) and new therapeutic strategies. This review aimed to present the prevalence, risk factors, current diagnosis, including artificial intelligence (AI) and machine learning approaches, future perspectives in combating HAIs causative bacteria (phage therapy, microbiome-based interventions, and vaccination), and HAIs surveillance strategies. Also, we discussed the latest findings regarding the relationships of AR with climate change and environmental pollution in the context of the One Health approach. Phage therapy is an emerging option that can offer an alternative to ineffective antibiotic treatments for antibiotic-resistant bacteria causing HAIs. Clinical trials dealing with vaccine development for resistant bacteria have yielded conflicting results. Two promising strategies, fecal microbiota transplantation and probiotic therapy, proved highly effective against recurrent Clostridium difficile infections and have been shown to reduce HAI incidence in hospitalized patients undergoing antibiotic therapy. Artificial intelligence and machine learning systems offer promising predictive capabilities in processing large volumes of clinical, microbiological, and patient data but require robust data integration. Our paper argues that HAIs are still a global challenge, requiring stringent IPC policies, computer vision, and AI-powered tools. Despite promising avenues like integrated One Health approaches, optimized phage therapy, microbiome-based interventions, and targeted vaccine development, several knowledge gaps in clinical efficacy, standardization, and pathogen complexity remain to be answered. © The Author(s) 2025.

Abstract

Antibiotic resistance represents a growing public health threat, with airborne drug-resistant strains being especially alarming due to their ease of transmission and association with severe respiratory infections. The respiratory microbiome plays a pivotal role in maintaining respiratory health, influencing the dynamics of antibiotic resistance among airborne pathogenic microorganisms. In this context, this review proposes the exploration of the complex interplay between the respiratory microbiota and antimicrobial resistance, highlighting the implications of microbiome diversity in health and disease. Moreover, strategies to mitigate antibiotic resistance, including stewardship programs, alternatives to traditional antibiotics, probiotics, microbiota restoration techniques, and nanotechnology-based therapeutic interventions, are critically presented, setting an updated framework of current management options. Therefore, through a better understanding of respiratory microbiome roles in antibiotic resistance, alongside emerging therapeutic strategies, this paper aims to shed light on how the global health challenges posed by multi-drug-resistant pathogens can be addressed. © 2025 by the authors.

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Abstract

The livestock industry has a harmful effect on the environment and contributes to climate change by producing significant amounts of greenhouse gases such as carbon dioxide (CO2), nitrous oxide (N2 O), methane (CH4), and ammonia (NH3). In line with this, the production and composting of manure is an important issue in livestock farming. During composting, the manure undergoes physical and nutritional profile changes, which can affect the environment. In this context, in the present study, we have investigated the dynamics of the chemical parameters such as pH, humidity +-(U%), organic matter (MO%), ammonium nitrogen N-NH4 (g/kg), nitrogen in the form of nitrite N-NO2 (g/kg), nitrate-nitrogen N–NO3 (g/kg) and phosphorus phosphate P-PO3-4 (g/kg) of organic manure from cattle and swine farms during composting. The results revealed a pH variation between 6.9 and 8.23 during composting, which is normal for compost, as mentioned in the literature. A high water content of over 90% was preserved in organic manure and compost because the samples were very liquid. Nutrient values showed increased ammonium and total inorganic nitrogen in cattle farms and one pig farm during composting. This study highlights a rise in nutrient values, especially of ammonium and total inorganic nitrogen in both cattle and pig farms, an aspect that reinforces the need to evaluate intervention strategies and develop a set of recommendations leading to the reduction of exposure risks and a subsequent reduction in human health impacts. © 2024, University of Zagreb - Faculty of Agriculture. All rights reserved.

Abstract

The current paper is focused on an emblematic medicinal species, Arnica montana, and on identification of the main threats it faces in the northeastern region of Romanian Carpathians. The species was investigated in 30 localities, in various habitats, characterized by different land management types, and different floristic composition of plant communities. The most important threat for A. montana in NE Carpathians was the habitat loss, mainly caused by abandonment of the traditional meadows management and natural succession of vegetation. Accumulated data underline the importance of management measures for A. montana, measures that have controlled and maintained the habitats of montane grasslands over time. The most suitable method of maintaining habitats in a favorable state of conservation could be represented by a combination of mowing and moderate intensity grazing while maintaining a level of fertilization as low as possible. Collection of flower heads for medicinal purposes must be avoided in small size populations.

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Abstract

Milk thistle (Silybum marianum (L.) Gaertn.) is among the top-selling botanicals used as a supportive treatment for liver diseases. Silymarin, a mixture of unique flavonolignan metabolites, is the main bioactive component of milk thistle. The biological activities of silymarin have been well described in the literature, and its use is considered safe and well-tolerated in appropriate doses. However, commercial preparations do not always contain the recommended concentrations of silymarin, failing to provide the expected therapeutic effect. While the poor quality of raw material may explain the low concentrations of silymarin, its deliberate removal is suspected to be an adulteration. Toxic contaminants and foreign matters were also detected in milk thistle preparations, raising serious health concerns. Standard methods for determination of silymarin components include thin-layer chromatography (TLC), high-performance thin-layer chromatography (HPTLC), and high-performance liquid chromatography (HPLC) with various detectors, but nuclear magnetic resonance (NMR) and ultra-high-performance liquid chromatography (UHPLC) have also been applied. This review surveys the extraction techniques of main milk thistle metabolites and the quality, efficacy, and safety of the derived food supplements. Advanced analytical authentication approaches are discussed with a focus on DNA barcoding and metabarcoding to complement orthogonal chemical characterization and fingerprinting of herbal products.

Abstract

Herbal medicines and preparations are widely used in healthcare systems globally, but concerns remain about their quality and safety. New herbal products are constantly being introduced to the market under varying regulatory frameworks, with no global consensus on their definition or characterization. These biologically active mixtures are sold through complex globalized value chains, which create concerns around contamination and profit-driven adulteration. Industry, academia, and regulatory bodies must collaborate to develop innovative strategies for the identification and authentication of botanicals and their preparations to ensure quality control. High-throughput sequencing (HTS) has significantly improved our understanding of the total species diversity within DNA mixtures. The standard concept of DNA barcoding has evolved over the last two decades to encompass genomic data more broadly. Recent research in DNA metabarcoding has focused on developing methods for quantifying herbal product ingredients, yielding meaningful results in a regulatory framework. Techniques, such as loop-mediated isothermal amplification (LAMP), DNA barcode-based Recombinase Polymerase Amplification (BAR-RPA), DNA barcoding coupled with High-Resolution Melting (Bar-HRM), and microfluidics-based methods, offer more affordable tests for the detection of target species. While target capture sequencing and genome skimming are considerably increasing the species identification resolution in challenging plant clades, ddPCR enables the quantification of DNA in samples and could be used to detect intended and unwanted ingredients in herbal medicines. Here, we explore the latest advances in emerging DNA-based technologies and the opportunities they provide as taxa detection tools for evaluating the safety and quality of dietary supplements and herbal medicines.

Abstract

In the current context of concern for the improvement and protection of environmental conditions, emphasis is placed on the provision of non-toxic, eco-friendly, renewable biomaterials to replace established chemical substances. Lythri herba is the aerial part of the plant species Lythrum salicaria L., known in the scientific literature especially for its content of tannins and total polyphenols, which highlight its antioxidant, hemostatic, antibacterial and antidiarrheal properties. Chitosan is a biopolymer widely used in industry and medicine due to its abundance in nature, its biodegradability, lack of toxicity and the ease with which it can be transformed into several basic forms (hydrogel, membrane, sponge). The aqueous solutions and membranes obtained in this study by merging these two natural resources were biologically tested in terms of genotoxicity (SOS-Chromo assay), hemolytic activity, thrombin generation activity and bacterial adhesion to reveal outwardly the lack of these properties and their use for medical purposes. The results of the current study attest to the absence of mutagenic and slight hemolyzing properties, thus supporting the possibility of using this extract and membrane in medical and pharmaceutical therapeutic practice. The surface parameters of membranes were examined and important influences at thrombin activity were found. Also, bacterial adhesion results showed a correlation between Lythri herba and chitosan concentrations and membranes' appearances (swelling, stability). The results show that the membranes could be a promising material for biomedical applications.

Abstract

BackgroundMilk thistle is one of the most popular hepatoprotectants, and is often sold in combination with other ingredients. Botanical supplements are known to be vulnerable to contamination and adulteration, and emerging technologies show promise to improve their quality control.MethodsUntargeted and semi-targeted metabolomics based on UHPLC-QTOF-ESI+MS techniques, UV spectrometry, and DNA metabarcoding using Illumina MiSeq were used to authenticate eighteen milk thistle botanical formulations (teas, capsules, tablets, emulsion).ResultsUntargeted metabolomics separated 217 molecules and by multivariate analysis the discrimination between the different preparations was established. The semi-targeted metabolomics focused on 63 phytochemicals, mainly silymarin flavonolignans and flavonoids, that may be considered as putative biomarkers of authenticity. All formulations contained molecules from silymarin complexes at different levels. The quantitative evaluation of silybins was done using in parallel UV spectrometry and UHPLC-QTOF-ESI+MS and their correlations were compared. DNA metabarcoding detected milk thistle in eleven out of sixteen retained preparations, whereas two others had incomplete evidence of milk thistle despite metabolomics validating specific metabolites, e.g., silymarin complex, identified and quantified in all samples. Meanwhile, the DNA metabarcoding provided insights into the total species composition allowing the interpretation of the results in a broad context.ConclusionOur study emphasizes that combining spectroscopic, chromatographic, and genetic techniques bring complementary information to guarantee the quality of the botanical formulations.

Abstract

This study identifies a source of heavy metal pollution in the Tarnia-Ostra area, Suceava County, Romania, using both chemical and biological techniques. Samples collected from the sterile dumps and the surrounding soil were digested and analyzed for total metals and arsenic by inductively coupled plasma-optical emission spectroscopy (ICP-OES) and atomic absorption spectrometry (AAS). High concentrations of heavy metals, such as copper (3.12 g/kg), arsenic (0.68 g/kg), lead (2.67 g/kg), zinc (0.43 g/kg) and barium (0.54 g/kg), found in that region, may pose a high risk to the environment and public health through the food chain. The large amounts of Cu, Fe, Pb, Zn and As of the collected samples diminished much the germination of wheat (Triticum aestivum L.), used in this study as a marker of toxicity. However, after the removal of the heavy metals by treating the aqueous extracts with sodium hydroxide solutions, the germination parameters returned to normal. The quantification of heavy metal levels by ICP-OES provides a better understanding of the potential ecological risks and contamination status of the tailings dumps. Instead, wheat germination tests can be reliable and inexpensive biological methods to prove the harmful effect of heavy metals and arsenic on plants.

Abstract

Traditional Chinese Medicine (TCM) is popular for the treatment of endometriosis, a complex gynecological disease that affects 10% of women globally. The growing market for TCMs has yielded a significant incentive for product adulteration, and although emerging technologies show promise to improve their quality control, many challenges remain. We tested the authenticity of two traditional Chinese herbal formulae used in women's healthcare for the treatment of endometriosis, known as Gui Zhi Fu Ling Wan (FL) and Ge Xia Zhu Yu Tang (GX). Dual-locus DNA metabarcoding analysis coupled with high-performance thin-layer chromatography (HPTLC) were used to authenticate 19 FL and six GX commercial herbal products, as well as three ad hoc prepared artificial mixtures. HPTLC was able to detect most of the expected ingredients via comparative component analysis. DNA metabarcoding was able to detect an unexpected species diversity in the products, including 38 unexpected taxa. Chromatography has a resolution for all species indirectly through the identification of marker compounds for the different species ingredients. Metabarcoding on the other hand yields an overview of species diversity in each sample, but interpretation of the results can be challenging. Detected species might not be present in quantities that matter, and without validated quantification, some detected species can be hard to interpret. Comparative analysis of the two analytical approaches also reveals that DNA for species might be absent or too fragmented to amplify as the relevant chemical marker compounds can be detected but no amplicons are assigned to the same species. Our study emphasizes that integrating DNA metabarcoding with phytochemical analysis brings valuable data for the comprehensive authentication of Traditional Chinese Medicines ensuring their quality and safe use.

Abstract

Sea buckthorn (Hippophae rhamnoides L.) (SB) is increasingly consumed worldwide as a food and food supplement. The remarkable richness in biologically active phytochemicals (polyphenols, carotenoids, sterols, vitamins) is responsible for its purported nutritional and health-promoting effects. Despite the considerable interest and high market demand for SB-based supplements, a limited number of studies report on the authentication of such commercially available products. Herein, untargeted metabolomics based on ultra-high-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UHPLC-QTOF-ESI+MS) were able to compare the phytochemical fingerprint of leaves, berries, and various categories of SB-berry herbal supplements (teas, capsules, tablets, liquids). By untargeted metabolomics, a multivariate discrimination analysis and a univariate approach (t-test and ANOVA) showed some putative authentication biomarkers for berries, e.g., xylitol, violaxanthin, tryptophan, quinic acid, quercetin-3-rutinoside. Significant dominant molecules were found for leaves: luteolin-5-glucoside, arginine, isorhamnetin 3-rutinoside, serotonin, and tocopherol. The univariate analysis showed discriminations between the different classes of food supplements using similar algorithms. Finally, eight molecules were selected and considered significant putative authentication biomarkers. Further studies will be focused on quantitative evaluation.

Abstract

Owing to its antibacterial, anti-inflammatory, and antioxidant activities, in the last few years, lavender essential oil (LVO) has been used in medical applications as a promising approach for treating infected wounds. However, the practical applicability of LVO is limited by its high volatility and storage stability. This study aimed to develop a novel hybrid hydrogel by combining phytic acid (PA)-crosslinked sodium alginate (SA) and poly(itaconic anhydride-co-3,9-divinyl-2,4,8,10-tetraoxaspiro[5.5] undecane (PITAU) and evaluate its potential effectiveness as an antibacterial wound dressing after incorporating LVO. The influence of the mass ratio between SA and PITAU on the properties and stability of hydrogels was investigated. After LVO loading, the effect of oil addition to hydrogels on their functional properties and associated structural changes was studied. FTIR analysis revealed that hydrogen bonding is the primary interaction mechanism between components in the hybrid hydrogels. The morphology was analyzed using SEM, evidencing a porosity dependent on the ratio between SA and PITAU, while LVO droplets were well dispersed in the polymer blend. The release of LVO from the hydrogels was determined using UV-VIS spectroscopy, indicating a sustained release over time, independent of the LVO concentration. In addition, the hybrid hydrogels were tested for their antioxidant properties and antimicrobial activity against Gram-positive and Gram-negative bacteria. Very good antimicrobial activity was obtained in the case of sample SA_PITAU3+LVO10% against S. aureus and C. albicans. Moreover, in vivo tests showed an increased antioxidant effect of the SA_PITAU3+LVO10% hydrogel compared to the oil-free scaffold that may aid in accelerating the healing process of wounds.

Abstract

Recently, there has been increased interest in the discovery of new natural herbal remedies for treating diabetes and inflammatory diseases. In this context, this work analyzed the antidiabetic and anti-inflammatory potential of Artemisia absinthium, Artemisia vulgaris and Trigonella foenum-graecum herbs, which have been studied less from this point of view. Therefore, extracts were prepared and processed using membrane technologies, micro- and ultrafiltration, to concentrate the biologically active principles. The polyphenol and flavone contents in the extracts were analyzed. The qualitative analysis of the polyphenolic compounds was performed via HPLC, identifying chlorogenic acid, rosmarinic acid and rutin in A. absinthium; chlorogenic acid, luteolin and rutin in A. vulgaris; and genistin in T. foenum-graecum. The antidiabetic activity of the extracts was analyzed by testing their ability to inhibit alpha-amylase and alpha-glucosidase, and the anti-inflammatory activity was analyzed by testing their ability to inhibit hyaluronidase and lipoxygenase. Thus, the concentrated extracts of T. foenum-graecum showed high inhibitory activity on a-amylase-IC50 = 3.22 +/- 0.3 mu g/mL-(compared with acarbose-IC50 = 3.5 +/- 0.18 mu g/mL) and high inhibitory activity on LOX-IC50 = 19.69 +/- 0.52 mu g/mL (compared with all standards used). The concentrated extract of A. vulgaris showed increased alpha-amylase inhibition activity-IC50 = 8.57 +/- 2.31 mu g/mL-compared to acarbose IC50 = 3.5 +/- 0.18 mu g/mL. The concentrated extract of A. absinthium showed pronounced LOX inhibition activity-IC50 = 19.71 +/- 0.79 mu g/mL-compared to ibuprofen-IC50 = 20.19 +/- 1.25 mu g/mL.

Abstract

Gerbillus is one of the most speciose genera among rodents, with ca. 51 recognized species. Previous attempts to reconstruct the evolutionary history of Gerbillus mainly relied on the mitochondrial cyt-b marker as a source of phylogenetic information. In this study, we utilize RAD-seq genomic data from 37 specimens representing 11 species to reconstruct the phylogenetic tree for Gerbillus, applying concatenation and coalescence methods. We identified four highly supported clades corresponding to the traditionally recognized subgenera: Dipodillus, Gerbillus, Hendecapleura and Monodia. Only two uncertain branches were detected in the resulting trees, with one leading to diversification of the main lineages in the genus, recognized by quartet sampling analysis as uncertain due to possible introgression. We also examined species boundaries for four pairs of sister taxa, including potentially new species from Morocco, using SNAPP. The results strongly supported a speciation model in which all taxa are treated as separate species. The dating analyses confirmed the Plio-Pleistocene diversification of the genus, with the uncertain branch coinciding with the beginning of aridification of the Sahara at the the PlioPleistocene boundary. This study aligns well with the earlier analyses based on the cyt-b marker, reaffirming its suitability as an adequate marker for estimating genetic diversity in Gerbillus.

Abstract

The genome edited crops and foods are commercially cultivated and marketed already at global level, rapidly expanding towards new applications and plant species, and successfully complementing the genetically modified ones. In the European Union, the genome edited plants have to follow the two-decade-old regulatory framework for genetically modified organisms. The decrease of both, number of notifications for field trials, and hectarage and countries commercially cultivating genetically modified plants, registered in the last decade in the European Union, has been closely followed by lower levels of awareness and concerns expressed by the European Union citizens about their use in farming and food production, as recorded by a long time series of Eurobarometers surveys. In contrast, in the four recent years only, the awareness about genome editing among the European Union citizens has significantly increased, reaching more than the half of the one about genetically modified products, along with the number of concerned European Union citizens about genome editing which also has doubled. The public opinion about genome edited crops and food products needs to be monitored further as it decisively influences the new regulatory framework to be proposed by the European Commission and therefore the extent to which the European consumer will benefit from the new biotechnologies. © The Author(s) 2024. All rights reserved.

Abstract

This study aims to demonstrate the effectiveness of silver nanoparticles (Ag NPs) on multidrug-resistant (MDR) Acinetobacter baumannii (AB) strains isolated from the clinical and aquatic environment. Three types of Ag NPs were investigated for their antimicrobial, antibiofilm, and antivirulence properties on a total number of 132 AB strains isolated in the same temporal sequence from intra-hospital infections (IHIs), wastewater (WW), and surface water (SW) samples between 2019 and 2022 from different Romanian locations and characterized at the phenotypic and genotypic levels. The comparative analysis of the antimicrobial resistance (AR) profiles according to the isolation source and the geographical location demonstrated a decrease in MDR level in AB recovered from WW samples in 2022 from north-eastern/central/southern regions (N-E/C-W/analyzed strains S): 87.5/60/32.5%. The AB strains were lecithinase, caseinase, amylase, and lipase producers, had variable biofilm formation ability, and belonged to six genotypes associated with the presence of different virulence genes (ompA, csuE, bap, and bfmS). The Ag NPs synthesized with the solvothermal method exhibited an inhibitory effect on microbial growth, the adherence capacity to the inert substratum, and on the production of soluble virulence factors. We report here the first description of a powerful antibacterial agent against MDR AB strains circulating between hospitals and anthropically polluted water in Romania. © 2023 by the authors.

Abstract

Background: Milk thistle is one of the most popular hepatoprotectants, and is often sold in combination with other ingredients. Botanical supplements are known to be vulnerable to contamination and adulteration, and emerging technologies show promise to improve their quality control. Methods: Untargeted and semi-targeted metabolomics based on UHPLC-QTOF-ESI+MS techniques, UV spectrometry, and DNA metabarcoding using Illumina MiSeq were used to authenticate eighteen milk thistle botanical formulations (teas, capsules, tablets, emulsion). Results: Untargeted metabolomics separated 217 molecules and by multivariate analysis the discrimination between the different preparations was established. The semi-targeted metabolomics focused on 63 phytochemicals, mainly silymarin flavonolignans and flavonoids, that may be considered as putative biomarkers of authenticity. All formulations contained molecules from silymarin complexes at different levels. The quantitative evaluation of silybins was done using in parallel UV spectrometry and UHPLC-QTOF-ESI+MS and their correlations were compared. DNA metabarcoding detected milk thistle in eleven out of sixteen retained preparations, whereas two others had incomplete evidence of milk thistle despite metabolomics validating specific metabolites, e.g., silymarin complex, identified and quantified in all samples. Meanwhile, the DNA metabarcoding provided insights into the total species composition allowing the interpretation of the results in a broad context. Conclusion: Our study emphasizes that combining spectroscopic, chromatographic, and genetic techniques bring complementary information to guarantee the quality of the botanical formulations. © 2023, The Author(s).

Abstract

Abstract

Herbal products comprise a wide spectrum of locally, nationally or internationally commercialized commodities. As these products have an increasingly important position in healthcare systems worldwide, a detailed product quality assessment is of crucial importance. For the quality evaluation of commercial herbal products, a wide range of methods were used, from simpler, quicker, and cost-effective HPTLC, to hyphenated methods with MS or NMR, where more precise quantification or specific structural information is required. Additionally, most of the methods have been coupled with chemometric tools, such as PCA, or PDA, for the multivariate analysis of the high amount of data generated by chromatograms, electropherograms or spectra. The chemical methods have revealed the widespread presence of low or variable quality herbal products in the marketplace. The majority of analytical investigations present major, qualitative and quantitative, inter-product variations of their chemical composition, ranging from missing ingredients, to strikingly and unnaturally high concentrations of some compounds. Moreover, the inter-batch quality variations were frequently reported, as well as the presence of some undesirable substances. The chemical analysis of herbal products is a vital component to raise the overall awareness of quality in the herbal market and generate a quality driven approach.

Abstract

Botanicals and derived herbal products have been long used in meeting primary healthcare needs around the world. They have gained increasing popularity in industrialized countries as complementary and alternative therapies to synthetic pharmaceuticals. Botanical preparations are however susceptible to contamination, adulteration, and substitution, that raise serious quality and public health safety concerns. The deliberate or accidental use of undeclared ingredients may occur throughout the entire value chain of the botanical product, from the cultivation or wild harvesting of the raw botanical material to the final packaged and marketed product. The current identification and quality inspection use tests to establish the identity, purity, and constituents of the herbal products. These tests most often utilize sensory and/or phytochemical inspection to detect species-specific characteristics or compounds, along with assays for toxic constituents such as heavy metals and toxins. However, marketed herbal products are very often highly processed and sold as complex mixtures. Classical analytical methods are often limited in their ability to identify the target plant species that should be present, and it is even more difficult to detect non-targeted species using these methods. This chapter presents a review of DNA barcoding and metabarcoding approaches to assess species diversity in herbal products and the use of these approaches in their quality control. © 2022 Elsevier Inc. All rights reserved.

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Abstract

Since 1996 till 2018, the global area cultivated with GM crops has increased 113-fold, making biotech crops one of the fastest adopted crop technology in the past decades. In the European Union, only two countries still cultivate one available transgenic crop event on minor hectarage. Moreover, the number of notifications for confined field trials has dramatically dropped in the last decade. All these are happening while the EU legislation on GM crops has come under severe criticism. The percentage of EU citizens concerned about the presence of GMOs in the environment has decreased from 30% (in 2002) to 19% (in 2011), while the level of concern about the use of GM ingredients in food or drinks has decreased from 63% (in 2005) to 27% (in 2019). The steadily increasing acceptance of the EU citizens of GMOs in the environment and food, as it was recorded by Eurobarometers, should additionally ease the way and support a positive change of the legal framework that regulates the GM crops' testing and commercial cultivation in the EU.

Abstract

Ginseng traditional medicines and food supplements are the globally top selling herbal products. Panax ginseng, Panax quinquefolius and Panax notoginseng are the main commercial ginseng species in herbal medicine. Prices of ginseng products vary widely based on the species, quality, and purity of the used ginseng, and this provides a strong driver for intentional adulteration. Our systematic literature search has reviewed the authenticity results of 507 ginseng-containing commercial herbal products sold in 12 countries scattered across six continents. The analysis of the botanical and chemical identity of all these products shows that 76% are authentic while 24% were reported as adulterated. The number of commercial products as well as the percentage of adulteration varies significantly between continents, being highest in South America (100%) and Australia (75%), and lower in Europe (35%), North America (23%), Asia (21%) and Africa (0%). At a national level, from the five countries for which more than 10 products have been successfully authenticated, the highest percentage of adulterated ginseng products were purchased from Taiwan (49%), followed by Italy (37%), China (21%), and USA (12%), while all products bought in South Korea were reported to be authentic. In most cases, labeled Panax species were substituted with other Panax species, but substitution of ginseng root, the medicinally recommended plant part, with leaves, stems or flowers was also reported. Efficient and practical authentication using biomarkers to distinguish the main ginseng varieties and secondary metabolite spectra for age determination are essential to combat adulteration in the global marketplace.

Abstract

Herbs and spices are some of the most vulnerable products in terms of fraud and adulteration in the food sector. Although standard analytical methods are accurate for quality control of specific lead or marker compounds, they cannot accurately assess the entire species composition of many marketed products. Complementary analytical approaches are thus often used for comprehensive screening of herbs and spices. In this study we evaluate DNA metabarcoding for the identification and authentication of 62 products, containing basil, oregano, and paprika collected from different retailers and importers in Norway. Our results show varying degrees of discrepancy between the constituent species and those listed on the product labels, despite high product authenticity. We suggest the false positives result from the sensitivity of DNA metabarcoding and filtering thresholds should be integrated into protocols to reduce false positives. Our results highlight how integrating DNA metabarcoding into the toolbox of analytical methods for quality control of fresh and/or processed plant-based food can improve product quality.

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Abstract

Endothelial dysfunction (impaired vasodilation, prothrombotic, proinflammatory and prooxidant states) is involved in the development of cardiovascular diseases (atherosclerosis, hypertension, heart failure, stroke) and vascular complications of diabetes mellitus. Reduced production of endothelium-derived relaxing factors, increased release of endothelium-derived constricting factors, prothrombotic molecules and inflammatory mediators, upregulation of growth factors and adhesion molecules play a crucial role in the development and progression of endothelial dysfunction. Phytochemicals belonging to different chemical classes (alkaloids, flavonoids, isoflavonoids, phenolic acids, terpenoids) were reported to prevent and/or reverse endothelial dysfunction in different experimental models. Many of them showed a multitarget activity which is a great advantage in tackling endothelial dysfunction and thereby, they may serve as lead compounds for novel drug development. Salvianolic acid B, caffeic, chlorogenic, ferulic and rosmarinic acids, resveratrol, luteolin are only some examples of phytochemicals modulating multiple mechanisms associated with endothelial dysfunction. Obviously, extensive clinical trials are needed to confirm the efficacy of phytochemicals in improving endothelial function and also their safety. In addition, formulation strategies to enhance bioavailability and afford an endothelium-targeted delivery should be explored.

Abstract

Chemical methods are the most important and widely used traditional plant identification techniques recommended by national and international pharmacopoeias. We have reviewed the successful use of different chemical methods for the botanical authentication of 2,386 commercial herbal products, sold in 37 countries spread over six continents. The majority of the analyzed products were reported to be authentic (73%) but more than a quarter proved to be adulterated (27%). At a national level, the number of products and the adulteration proportions varied very widely. Yet, the adulteration reported for the four countries, from which more than 100 commercial products were purchased and their botanical ingredients chemically authenticated, was 37% (United Kingdom), 31% (Italy), 27% (United States), and 21% (China). Simple or hyphenated chemical analytical techniques have identified the total absence of labeled botanical ingredients, substitution with closely related or unrelated species, the use of biological filler material, and the hidden presence of regulated, forbidden or allergenic species. Additionally, affecting the safety and efficacy of the commercial herbal products, other low quality aspects were reported: considerable variability of the labeled metabolic profile and/or phytochemical content, significant product-to-product variation of botanical ingredients or even between batches by the same manufacturer, and misleading quality and quantity label claims. Choosing an appropriate chemical technique can be the only possibility for assessing the botanical authenticity of samples which have lost their diagnostic microscopic characteristics or were processed so that DNA cannot be adequately recovered.

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Abstract

The cross-compliance mechanism of the European Union (EU)'s common agricultural policy (CAP) makes the approval of the direct payments to the European farmers subject to compliance with the requirement to maintain the land in good agricultural and environmental condition. One of the obligations of the Romanian land owners and farmers is to avoid the installation of unwanted vegetation on their land plots. This vegetation is represented by some species of herbaceous or woody plants, annual or perennial, that spontaneously invade the agricultural lands, diminishing the production capacity of the cultivated plants. Included in this category are 10 meadow weeds, without fodder value or even toxic to animals: Arctium lappa L., Carduus nutans L., Conium maculatum L., Eryngium campestre L., Euphorbia cyparissias L., Pteridium aquilinum (L.) Kuhn, Rumex acetosella L., Veratrum album L., Xanthium spinosum L., and Xanthium strumarium L. Various and multiple uses in traditional medicine of these meadow weed species have been reported for Romania and other nine neighboring East European countries, i.e. Bosnia and Herzegovina, Bulgaria, Czech Republic, Estonia, Kosovo, Russia, Turkey, Serbia, and Ukraine. For A. lappa were recorded the highest number of ethnomedicinal uses, in the largest number of East European countries, including Romania. C. maculatum and V. album are not recommended for human consumption but can be further investigated as potential sources of pharmaceutically active compounds. Once removed by landowners and farmers from their land, the raw plant material of these 10 species become readily and easily available to the Romanian local communities and the industry of herbal food supplements, while the biodiversity of the agro-ecosystems is maintained.

Abstract

Herbal products are marketed and used around the globe for their claimed or expected health benefits, but their increasing demand has resulted in a proportionally increase of their accidental contamination or intentional adulteration, as already confirmed with DNA-based methods. Microscopy is a traditional pharmacopoeial method used for plant identification and we systematically searched for peer-reviewed publications to document its potential and limitations to authenticate herbal medicines and food supplements commercially available on the global market. The overall authenticity of 508 microscopically authenticated herbal products, sold in 13 countries, was 59%, while the rest of 41% were found to be adulterated. This problem was extending over all continents. At the national level, there were conspicuous differences, even between neighboring countries. These microscopically authenticated commercial herbal products confirm that different magnifying instruments can be used to authenticate crude or processed herbal products traded in the global marketplace. The reviewed publications report the successful use of different magnifying instruments, single or in combinations with a second one, with or without a chemical or DNA-based technique. Microscopy is therefore a rapid and cost-efficient method, and can cope with mixtures and impurities. However, it has limited applicability for highly processed samples. Microscopic authentication of commercial herbal products will therefore contribute to raise public awareness for the extent of adulteration and the need to safeguard consumer safety against the challenges of globalization.

Abstract

The emergence of multidrug resistant pathogens is a great challenge to the medical field and a major global health threat requiring novel therapeutic strategies. Among plant products, essential oils have significant antimicrobial properties that make them promising agents in the fight against drug resistant human pathogens. The aim of the present review was to highlight the most important essential oil-based antimicrobial strategies as revealed by recent studies. Synergistic interactions between essential oils or their bioactive compounds in combination with known antibiotics are presented. Also, nanoformulation approaches to boost the antimicrobial activity of essential oils are reviewed in terms of bioefficiency, stability and design of the nanostructured delivery systems. The focus was mainly put on the antimicrobial activity against multi-drug resistant pathogens, also called ESKAPE organisms (Enterococcusspp.,Staphylococcus aureus,Klebsiellaspp.,Acinetobacter baumannii,Pseudomonas aeruginosa, andEnterobacterspp.). Thus, essential oils in combinatorial and nano-based strategies may cope with infections caused by drug-resistant bacteria and may offer possibilities for reducing antibiotic use. Research on thein vivoefficacy and safety of such strategies is required for further clinical antimicrobial chemotherapy. In this regard, the understanding of the interactions between essential oil-based strategies and biological interface is essential.

Abstract

We report on the genomic characterization of 47 multi-drug resistant, carbapenem resistant and ESBL-producing K. pneumoniae isolates from the influent (I) and effluent (E) of three wastewater treatment plants (WWTPs) and from Romanian hospital units which are discharging the wastewater in the sampled WWTPs. The K. pneumoniae whole genome sequences were analyzed for antibiotic resistance genes (ARGs), virulence genes and sequence types (STs) in order to compare their distribution in C, I and E samples. Both clinical and environmental samples harbored prevalent and widely distributed ESBL genes, i.e. blaSHV, blaOXA, blaTEM and blaCTX M. The most prevalent carbapenemase genes were blaNDM-1, blaOXA-48 and blaKPC-2. They were found in all types of isolates, while blaOXA-162, a rare blaOXA-48 variant, was found exclusively in water samples. A higher diversity of carbapenemases genes was seen in wastewater isolates. The aminoglycoside modifying enzymes (AME) genes found in all types of samples were aac(6’), ant(2’’)Ia, aph(3’), aaD, aac(3) and aph(6). Quinolone resistance gene qnrS1 and the multi-drug resistance oqxA/B pump gene were found in all samples, while qnrD and qnrB were associated to aquatic isolates. The antiseptics resistance gene qacEdelta1 was found in all samples, while qacE was detected exclusively in the clinical ones. Trimethroprim-sulfamethoxazole (dfrA, sul1 and sul2), tetracyclines (tetA and tetD) and fosfomycin (fosA6, known to be located on a transpozon) resistance genes were found in all samples, while for choramphenicol and macrolides some ARGs were detected in all samples (catA1 and catB3 / mphA), while other (catA2, cmIA5 and aac(6’)Ib / mphE and msrE) only in wastewater samples. The rifampin resistance genes arr2 and 3 (both carried by class I integrons) were detected only in water samples. The highly prevalent ARGs preferentially associating with aquatic versus clinical samples could ascribe potential markers for the aquatic (blaSHV-145, qacEdelta1, sul1, aadA1, aadA2) and clinical (blaOXA-1, blaSHV-106,blaTEM-150, aac(3)Iia, dfrA14, oqxA10; oqxB17,catB3, tetD) reservoirs of AR. Moreover, some ARGs (oqxA10; blaSHV-145; blaSHV-100, aac(6’)Il, aph(3’) VI, armA, arr2, cmlA5, blaCMY-4, mphE, msrE, oqxB13, blaOXA-10) showing decreased prevalence in influent versus effluent wastewater samples could be used as markers for the efficiency of the WWTPs in eliminating AR bacteria and ARGs. The highest number of virulence genes (75) was recorded for the I samples, while for E and C samples it was reduced to half. The most prevalent belong to three functional groups: adherence (fim genes), iron acquisition (ent, fep, fyu, irp and ybt genes) and the secretion system (omp genes). However, none of the genes associated with hypervirulent K. pneumoniae have been found. A total of 14 STs were identified. The most prevalent clones were ST101, ST219 in clinical samples and ST258, ST395 in aquatic isolates. These STs were also the most frequently associated with integrons. ST45 and ST485 were exclusively associated with I samples, ST11, ST35, ST364 with E and ST1564 with C samples. The less frequent ST17 and ST307 aquatic isolates harbored blaOXA-162, which was co-expressed in our strains with blaCTX-M-15 and blaOXA-1. © 2020 Surleac et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

The cross-compliance mechanism of the European Union (EU)'s common agricultural policy (CAP) makes the approval of the direct payments to the European farmers subject to compliance with the requirement to maintain the land in good agricultural and environmental condition. One of the obligations of the Romanian land owners and farmers is to avoid the installation of unwanted vegetation on their land plots. This vegetation is represented by some species of herbaceous or woody plants, annual or perennial, that spontaneously invade the agricultural lands, diminishing the production capacity of the cultivated plants. Included in this category are 10 meadow weeds, without fodder value or even toxic to animals: Arctium lappa L., Carduus nutans L., Conium maculatum L., Eryngium campestre L., Euphorbia cyparissias L., Pteridium aquilinum (L.) Kuhn, Rumex acetosella L., Veratrum album L., Xanthium spinosum L., and Xanthium strumarium L. Various and multiple uses in traditional medicine of these meadow weed species have been reported for Romania and other nine neighboring East European countries, i.e. Bosnia and Herzegovina, Bulgaria, Czech Republic, Estonia, Kosovo, Russia, Turkey, Serbia, and Ukraine. For A. lappa were recorded the highest number of ethnomedicinal uses, in the largest number of East European countries, including Romania. C. maculatum and V. album are not recommended for human consumption but can be further investigated as potential sources of pharmaceutically active compounds. Once removed by landowners and farmers from their land, the raw plant material of these 10 species become readily and easily available to the Romanian local communities and the industry of herbal food supplements, while the biodiversity of the agro-ecosystems is maintained. © Copyright © 2020 Grosu and Ichim.

Abstract

Separation of naphtodianthrones (NTs) from Hypericum perforatum L. (aerial part of St. John’s Wort) is still topical due to some hard-to-beat medicinal attributes of these bioactive compounds. Unfortunately, their low bioavailability (0.06%–0.4%) complicates the extraction process. Therefore, developing straightforward and lower-cost methodologies for NT separation is still a priority. In support of this purpose, for preparing NT formulations from flowers and leaves of wild St. John’s Wort (hyperici herba), a cutoff preparative methodology is described herein. Combining Soxhlet extraction and reflux extraction, some concentrated and rather pure NT ethanolic-based formulations without chlorophyl and grease were obtained.

Abstract

Romania was the third country in Europe and the tenth in the world, to commercially adopt genetically modified crops in 1999, only 3years after they were first marketed globally. Half a million hectares of transgenic herbicide resistant soybean and insect resistant maize were grown in Romania during an uninterrupted 17-year period. After several years of continued declining area, the commercial cultivation of transgenic plants recently ended. The commercial cultivation of transgenic crops in Romania remains legally and technically possible, according to the EU and national regulations. However, the declining area cultivated with these crops in Romania seems to be the result of farmers' conscious decision, while waiting for more profitable genetically modified crops to become available that better fit their needs. This expectation would be a logical result of the EU GMO opt-out Directive 2015/412 and the advent of the new plant breeding techniques. The GMO opt-out mechanism is still expected to unblock the EU authorization process after the large majority of the EU member states have already prohibited the cultivation of genetically modified organisms in their territory. As the new plant breeding techniques offer significant technical and economic advantages, they could be rapidly adopted by commercial breeders and farmers outside Europe. The Court of Justice of the European Union ruled that plants obtained with the new gene editing techniques must go through the same authorization procedure as transgenic plants. This decision is expected to delay the approval and availability of these new plant varieties on the EU market and their commercial cultivation.

Abstract

The herbal products, sold worldwide as medicines or foods, are perceived as low risk because they are considered natural and thus safe. The quality of these products is ineffectively regulated and controlled. The growing evidence for their lack of authenticity is causing deep concern, but the scale of this phenomenon at the global, continental or national scale remains unknown. We analyzed data reporting the authenticity, as detected with DNA-based methods, of 5,957 commercial herbal products sold in 37 countries, distributed in all six inhabited continents. Our global survey shows that a substantial proportion (27%) of the herbal products commercialized in the global marketplace is adulterated when their content was tested against their labeled, claimed ingredient species. The adulterated herbal products are distributed across all continents and regions. The proportion of adulterated products varies significantly among continents, being highest in Australia (79%), South America (67%), lower in Europe (47%), North America (33%), Africa (27%) and the lowest in Asia (23%). The commercial HPs' authenticity among the 37 countries included in our global analysis ranges between 0 and 100% from the total number of product reported for each specific national marketplace. For 9 countries, more than 100 products were successfully DNA-based authenticated and reported. From these countries, the highest percentage of adulterated commercial HPs was reported for Brazil (68%), followed distantly by Taiwan (32%), India (31%), USA (29%), followed closely by Malaysia (24%), Japan (23%), South Korea (23%), Thailand (20%), and China (19%). Our results confirm the large-scale presence of adulterated herbal products throughout the global market. The adulterated herbal products contain undeclared contaminant, substitute, and filler species, or none of the labeled species, which all may be accidental or intentional, economically-motivated and fraudulent. Due to the ever-increasing analytical sensitivity of the high throughput DNA sequencing, increasingly used for the untargeted, simultaneous multi-taxa identification, the proportion of adulterated HPs detected on the global market is expected to increase. In the context of the increasing demand for HPs, the limited supply of raw materials derived from many plant species, some of which being already nationally or internationally protected and having various degrees of trade restrictions, adds up to the differences and discrepancies between national HPs' regulatory frameworks and further increases the risks of adulteration of many types of herbal products. The globally widespread adulteration is a serious threat to consumers' well-being and safety, in spite of herbal products' claimed or expected health benefits.

Abstract

Ayurveda is one of the oldest systems of medicine in the world, but the growing commercial interest in Ayurveda based products has increased the incentive for adulteration and substitution within this herbal market. Fraudulent practices such as the use of undeclared fillers and use of other species of inferior quality is driven both by the increased as well as insufficient supply capacity of especially wild plant species. Developing novel strategies to exhaustively assess and monitor both the quality of raw materials and final marketed herbal products is a challenge in herbal pharmacovigilance. Seventy-nine Ayurvedic herbal products sold as tablets, capsules, powders, and extracts were randomly purchased via e-commerce and pharmacies across Europe, and DNA metabarcoding was used to assess the ability of this method to authenticate these products. Our analysis reveals that only two out of 12 single ingredient products contained only one species as labeled, eight out of 27 multiple ingredient products contained none of the species listed on the label, and the remaining 19 products contained 1 to 5 of the species listed on the label along with many other species not specified on the label. The fidelity for single ingredient products was 67%, the overall ingredient fidelity for multi ingredient products was 21%, and for all products 24%. The low level of fidelity raises concerns about the reliability of the products, and detection of threatened species raises further concerns about illegal plant trade. The study highlights the necessity for quality control of the marketed herbal products and shows that DNA metabarcoding is an effective analytical approach to authenticate complex multi ingredient herbal products. However, effort needs to be done to standardize the protocols for DNA metabarcoding before this approach can be implemented as routine analytical approaches for plant identification, and approved for use in regulated procedures.

Abstract

In many European countries Arnica montana is decreasing due to intensification or abandonment of traditional extensive land use and thus is considered endangered. In Romania the species is also decreasing due to excessive collecting for pharmaceutical and cosmetic purposes, but it is still relatively common in montane nutrient-poor grasslands and successional vegetation of forest clearings on acidic soil. In this study we analysed habitats and population structure of A. montana in the Romanian NE Carpathians. We asked for differences in population structure between habitat types and how population structure is related to environmental conditions. We investigated population structure and habitat characteristics in 25 populations of A. montana on three 1 m x 1 m-plots each (total of 75 plots). The plot-based assessment of the population structure included the numbers of rosettes (rosette density), flower heads (flower head density), flowering rosettes and flower stems per flowering rosette. From these variables we calculated the proportions of flowering and vegetative rosettes and the number of flower heads per flowering rosette. Habitats were characterised using climatic data (including elevation), land use types (grazing, mowing or abandonment), vegetation types, soil measurements (pH, several nutrient elements, organic matter content) and N supply on basis of plant indicator values. Vegetation types were defined by using hierarchical clustering and linked to phytosociological syntaxa based on diagnostic species. Observed habitat types included Mountain hay meadows, Species-rich Nardus grasslands and Alpine and subalpine heathlands according to the European Habitats Directive. DCA analysis was used for detecting floristic gradients, which were subsequently correlated to environmental conditions. Relationships between rosette density, proportion of vegetative rosettes, flower head density and number of flower heads per flowering rosette and environmental variables were assessed using GLMs. Five syntaxa with A. montana were identified: Festuco rubrae-Agrostietum capillaris nardetosum strictae, Scorzonero roseae-Festucetum nigricantis, Violo declinatae-Nardetum strictae, Cetrario-Vaccinietum gaultherioidis and Campanulo abietinae-Vaccinietum myrtilli. While vegetation composition was mainly correlated with altitude and soil variables such as pH and organic matter content, altitude was an important predictor for the proportion of flowering rosettes of A. montana, which decreased with altitude. Rosette density of A. montana varied between habitat types and was highest in Mountain hay meadows and lowest in plant communities dominated by tall plants with high cover. Organic matter content of the soil was the only important (negative) predictor for the number of flower heads per rosette, while a hump-shaped correlation was found between the density of inflorescences and the soil pH, with a maximum around pH 4.5. Grassland management was also an important factor for rosette density of A. montana, suggesting that maintaining and promoting traditional land use practices will be the most effective measure for the species' conservation.

Abstract

Ajuga genevensis and Ajuga reptans are medicinal plants often used as remedies in the Romanian traditional medicine for their benefic properties. They are also credited with astringent, tonic and diuretic activities. Although these species are common for the wild flora in our country, our investigation was conducted on specimens harvested from the experimental fields of Stejarul Biological Research Centre, Piatra Neamt, Romania. Their introduction in culture as a source of bioactive compounds is intended for pharmaceutical purposes. Their chemical profile was assessed on hydro-alcoholic extracts by spectrophotometry and RP-UPLC techniques. The in vitro biological effects of the samples were investigated against lipoxygenase, butyryl- and acetylcholinesterase. For the in vivo biological activities, the extracts (25 and 75 mg/kg bw) were administered in a 6-hydroxidopamine Parkinsonian rat model. The results confirmed the taxonomic and chemical variability between the investigated samples. In terms of compound profile, the polyphenols (flavonoid and polyphenolic acids) were in a higher amount in Ajuga genevensis (168.3 mg % and 230.4 mg % respectively), whereas Ajuga reptans contained higher quantities of iridoids (1860 mg % as compared to 1250 mg %). Harpagide and its acetylated derivative were the main iridoids identified in both extracts. The biological activity confirmed the antioxidant and protective properties of both extracts. Moreover, the anti-amnesic activity was significantly increased for Ajuga reptans as compared to Ajuga genevensis. Such results confirm a direct correlation between the concentration or iridoids and neuroprotective capacity of the extract obtained from A. reptans.

Abstract

Humankind draws important benefits from large-scale ecological processes termed ecosystem services, yet the status of several of them is declining. Reliable monitoring methods are essential for tracking the status of ecosystem services. Predation is the mainstay of natural pest control, a key ecosystem service. We used green plasticine caterpillars to monitor predation pressure, and to obtain baseline data on predator activity in transgenic Bt versus non-Bt maize fields in Old and New World countries. Predation pressure was measured at ground and canopy levels using an identical, small-plot experimental design in four European countries (Denmark, Slovakia, Romania and Italy) and Argentina. Total predation rate in maize was 11.7%d(-1) (min. 7.2%d(-1) in Argentina, max. 29.0%d(-1) in Romania). Artificial caterpillars were attacked both by invertebrates (mostly chewing insects with 42.0% of the attack marks, and ants with 7.1%, but also predatory and parasitoid wasps, spiders and slugs), and vertebrates (small mammals 25.5%, and birds 20.2%). Total predation at ground level (15.7%d(-1)) was significantly higher than in maize canopies (6.0%d(-1)) in all countries, except Argentina. We found no significant differences between predator pressure in Bt versus non-Bt maize plots. The artificial caterpillar method provided comparable, quantitative data on predation intensity, and proved to be suitable for monitoring natural pest control. This method usefully expands the existing toolkit by directly measuring ecological function rather than structure.

Abstract

Heavy metals are destructive environmental stressors that greatly reduce plant growth and productivity. Here, the content of some heavy metals (Fe, Cu, Zn and Mn) and mineral nutrients (Ca, Mg and K) in silver fir (Abies alba Mill.) and European beech (Fagus sylvatica L.) wood from two sites in the Tarnipa mining area (North-Est Romania) was analyzed. The first site (no. 1) is located near a big tailings dump, whereas the other one (no. 2) is situated 6 km South, in a less polluted area. The purpose of this work was to evaluate the present-date levels of such elements in the investigated biological materials and to compare them. Measurements of heavy metal content in wood were performed with an AAS flame spectrometer, whereas an ICP-OES spectrometer was used for soil and tailings dump samples. The Mn content was higher in the site no. 1 located near the dump, for both species, but the difference was statistically significant only for the European beech trees. The Zn content of samples from the site no. 1 was lower than that corresponding to the second one (no. 2) for both species, but significant for the silver fir only. However, there was not found a significant difference in respect to both iron and copper in the samples collected from the two species and two different sites. Furthermore, the content of Ca and Mg of both species was found higher for the site no. 2, suggesting a better state of nutrition of the investigated trees than of those located near the tailings dump. The content of calcium and magnesium of silver fir was negatively correlated with the diameter of the trees collected from both sites.

Abstract

Despite recent fruitful attempts to elucidate microbial biogeography in more detail, knowledge of distribution still lags behind for dinophytes. Evolutionary phenomena, such as cryptic speciation and modification due to the environment, hamper reliable conclusions about the distribution of this important plankton group. We combined newly collected samples from the Black Sea (ten new strains from three localities) with occurrence data, which have been gathered extensively over the past decade, in order to provide the first global distribution maps of four specific ribotypes assigned to the Scrippsiella lineage (Thoracosphaeraceae, Peridiniales) collected at a total of 39 sites. They showed a wide, partly overlapping distribution and shared the presence primarily at the coastal localities. Differences in abundance of specific ribotypes were observed, but the ribotype corresponding to the globally most frequently encountered species Scrippsiella acuminata has not yet been found in the Black Sea. We discuss the significance of DNA-based records for distribution maps particularly of unicellular organisms such as dinophytes. Based on a collective approach as exemplified in our study, we may start to understand in detail the ecological basis and the dynamics of the individual colonisation/invasion events, species establishment and consequent distribution in the microbiome, all of which have been changing drastically due to the ongoing climate change.

Abstract

Thymus species are important aromatic, medicinal and culinary plants with a significant economic value. This study evaluated for the first time the chemical composition and in vitro bioactivities of the essential oils (EOs) from five Moldavian Thymus species (T. vulgaris, T. x citriodorus, T. calcareus) and cultivars (T. vulgaris 'Faustini', T. citriodorus 'Aureus'). The main compounds in Thymus EOs were: thymol in T. vulgaris and T. calcareus EOs (55.44% and 55.45%, respectively), lavandulol in T. x citriodorus EO (54.27%), and geraniol in T. citriodorus 'Aureus' and T. vulgaris 'Faustini' EOs (60.31% and 31.45%, respectively). T. vulgaris and T. calcareus EOs showed the most potent antioxidant activities (EC50=0.003 mg/mL in ABTS radical cation scavenging assay) and exhibited significant inhibitory effects against aflatoxin-producing Aspergillus flavus fungus (MIC=0.25 mu L/mL). At doses that provided micromolar concentrations of thymol, T. vulgaris and T. calcareus EOs acted genoprotective at preventive and interventional levels against H2O2-induced genomic damage in V79 cells, the former being more active (6.21% and 5.52% vs. 25.13% and 7.26% tail DNA in pre- and post-treatment protocols, respectively). The genoprotective effects may be ascribed to antioxidant potential and, possibly, to stimulation of DNA repair processes. The Moldavian Thymus species are valuable resources of bioactive EOs for pharmaceutical and food industries (T. vulgaris, T.calcareus) but also for flavor industry and perfumery (T. x citriodorus, T. citriodorus 'Aureus', T. vulgaris 'Faustini').

Abstract

IntroductionHerbal medicines play an important role globally in the health care sector and in industrialised countries they are often considered as an alternative to mono-substance medicines. Current quality and authentication assessment methods rely mainly on morphology and analytical phytochemistry-based methods detailed in pharmacopoeias. Herbal products however are often highly processed with numerous ingredients, and even if these analytical methods are accurate for quality control of specific lead or marker compounds, they are of limited suitability for the authentication of biological ingredients. ObjectiveTo review the benefits and limitations of DNA barcoding and metabarcoding in complementing current herbal product authentication. MethodRecent literature relating to DNA based authentication of medicinal plants, herbal medicines and products are summarised to provide a basic understanding of how DNA barcoding and metabarcoding can be applied to this field. ResultsDifferent methods of quality control and authentication have varying resolution and usefulness along the value chain of these products. DNA barcoding can be used for authenticating products based on single herbal ingredients and DNA metabarcoding for assessment of species diversity in processed products, and both methods should be used in combination with appropriate hyphenated chemical methods for quality control. ConclusionsDNA barcoding and metabarcoding have potential in the context of quality control of both well and poorly regulated supply systems. Standardisation of protocols for DNA barcoding and DNA sequence-based identification are necessary before DNA-based biological methods can be implemented as routine analytical approaches and approved by the competent authorities for use in regulated procedures. (c) 2017 The Authors. Phytochemical Analysis Published by John Wiley & Sons Ltd. Herbal medicines play an important role globally in the health care sector and in industrialised countries they are often considered as an alternative to mono-substance medicines. Different methods have varying resolution and usefulness along the value chain of these products. DNA barcoding can be used for authenticating products based on single herbal ingredients and DNA metabarcoding for assessment of species diversity in processed products, and both methods should be used in combination with appropriate hyphenated chemical methods for quality control.

Abstract

Heavy metal pollution of ecosystems is of great concern, due to the persistence of metals in ecosystem. In this regard, the mountainous pastures of Romanian Carpathians, especially those in the areas of abandoned mines, need particular attention. The aim of this work was to assess the effect of heavy metals and metalloids exposure on cuticular wax composition of tall fescue leaves (Festuca arundinacea). Therefore, we have particularly investigated the variations of long chain hydrocarbon fractions, which are acknowledged to act as a protection for plants especially in toxic environments. The exposure experiments were conducted in triplicates for arsenic and the following metal ions: Ba2+, Cu2+, Fe2+ and Pb2+. The 0.5 mM aqueous solutions of their corresponding salts were used as treatment solutions. Triplicate experiments were also performed for control samples using distilled water as exposure media. The plantlets were kept for growing in controlled environment for 15 days followed by n-hexane hydrocarbon fraction extraction from 0.5 g of sampled leaves. The obtained extracts were semi-quantitatively analyzed (identification followed by peak area measurement) through an optimized method based on gas-chromatography coupled with mass spectrometry. We focused on the measurement of several long chain n-alkanes with the following number of carbon atoms: C21-C31. Significant differences were obtained between results performed for Ba2+, Pb2+ and Fe2+, when compared to AsO43- and Cu2+, which may suggest multiple mechanisms through which the tested plants could develop and adapt when exposed to various chemicals characterized by different degrees of toxicity.

Abstract

Background: Differences in regulatory policies between countries as well as a lack of appropriate standardized methods for the authentication and quality control of herbal products directly impact their quality and safety. Echinacea products are among the top-selling herbal products in Europe and the United States with indications for a broad range of ailments. The increased use of Echinacea species has led to concerns about adulterated products resulting from challenges in morphology-based identification, due to overlapping morphological variation, frequent hybridization between species, and deliberate adulteration. Purpose: This study addressed the need for a novel analytical strategy in the authentication of herbal products. Methods: A combination of high performance thin layer chromatography (HPTLC) and DNA metabarcoding was employed. Fifty-three Echinacea herbal products marketed across Europe were tested to evaluate the accuracy of these methods in plant identification and their potential for detecting substitutes, adulterants and other unreported plant constituents. Results: HPTLC provides high resolution in the detection of Echinacea phytochemical target compounds, but does not offer information on the other species within the product. Alternatively, we showed that the limitation of HPTLC in detecting non-targeted species can be overcome by the complementary use of DNA metabarcoding. Using DNA metabarcoding, Echinacea species were detected in 34 out of the 38 retained products (89%), but with a lack of discriminatory resolution at the species level due to the low level of molecular divergence within the Echinacea genus. All of the tested herbal products showed considerable discrepancies between ingredients listed on the label and the ones detected using DNA metabarcoding, registering an overall ingredient fidelity of only 43%. Conclusion: The results confirm that DNA metabarcoding can be used to test for the presence of Echinacea species and simultaneously to detect other species present in even highly processed and multi-ingredient herbal products.

Abstract

In consideration of the hazards associated with the presence of the textile azo-dye and their biotransformation products in the environment, the goal of this work was to study bioremediation process by the yeast strain Pichia kudriavzevii CR-Y103 related to the ability to degrade and detoxify the sulfonated Reactive Orange 16 azo-dye. In experimental conditions, the optimal inoculum/dye concentration ratio required for complete decolorization (100%) of culture medium and biomass within 24 h has been 1 g L-1 yeast cell (dry weight)/50 mg L-1 Reactive Orange 16. In the presence of 400 mg L-1 of Reactive Orange 16 (RO16), 95% of the dye was removed after 72 h of incubation. Also, the yeast strain could decolorize other eight textile dyes (56.48-99.98% decolorization within 24 h). NADH-DCIP reductase and azo reductase activities were significantly increased (ca. 5.4 times and ca. 37 times, respectively) during the decolorization process. UV-VIS spectra, high-performance liquid chromatography (HPLC), and Fourier transform infrared spectroscopy (FTIR) analysis confirmed the presence of new biotransformation products in extracted metabolites, highlighting the partial biodegradation of the dye by the new yeast isolate. The phytotoxicity evaluation strongly supported the decreased toxicity of biodegraded products as minor inhibition on germination (%), root and shoots elongation of T. pratense L. and T. aestivum L. seedlings. Increasing of mitotic index value and decreasing the frequency of chromosomal aberrations in tested plant meristem cells treated with biodegraded products, compared with RO16 treatment (500 ppm), confirmed their slightly toxic nature. A cell viability assay also confirmed the reduced toxicity of biodegraded products on healthy monkey kidney cells (Vero cells).

Abstract

Abstract

Lavandula species are one of the most useful aromatic and medicinal plants with a great economic value for pharmaceutical, food and flavor industries, cosmetics, perfumery, and aromatherapy. The pleasant fragrance, physiological effects, and commercial significance are due to the their essential oils, with a highly variable composition dependent on the genetic, environmental and processing factors. This paper provides a systematic view on the chemistry of lavender essential oils especially considering the studies in the last 15 years. The review presents the characteristic chemical constituents and chemotypes of seventeen Lavandula species. The intra- and interspecific variability is primarily discussed dependent on the geographic area, onto-and morphogenetic factors, and extraction methods. In addition, biosynthesis of lavender terpenes is presented.

Abstract

Many herbal products have a long history of use, but there are increasing concerns over product efficacy, safety and quality in the wake of recent cases exposing discrepancies between labeling and constituents. When it comes to St. John's wort (Hypericum perforatum L.) herbal products, there is limited oversight, frequent off-label use and insufficient monitoring of adverse drug reactions. In this study, we use amplicon metabarcoding (AMB) to authenticate 78 H. perforatum herbal products and evaluate its ability to detect substitution compared to standard methods using thin-layer chromatography (TLC) and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Hypericum perforatum was detected in 68% of the products using AMB. Furthermore, AMB detected incongruence between constituent species and those listed on the label in all products. Neither TLC nor HPLC-MS could be used to unambiguously identify H. perforatum. They are accurate methods for authenticating presence of the target compounds, but have limited efficiency in detecting infrageneric substitution and do not yield any information on other plant ingredients in the products. Random post-marketing AMB of herbal products by regulatory agencies could raise awareness among consumers of substitution and would provide an incentive to manufacturers to increase quality control from raw ingredients to commercialized products.

Abstract

The objectives of this study were to define the phenolic profile, antioxidant, antimicrobial and anti-inflammatory properties of extracts from Betonica officinalis and Salvia officinalis herbs used in traditional medicine. The results indicated that the extract inhibits scavenges stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation radicals. In the present study, Salvia officinalis was potent antibacterial agent against Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. The extracts inhibited cyclooxygenase (COX-1) and denaturation of protein, which suggests these species might be a potential source of plant-derived anti-inflammatory substances. Our study, for the first time, revealed the antimicrobial and anti-inflammatory activity of the Betonica officinalis herb. The results of this study could be helpful to develop medicinal preparations and uses of this species as an antimicrobial, anti-inflammatory and wound healing agent.

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Studying herbal products derived from local and traditional knowledge and their value chains is one of the main challenges in ethnopharmacology. The majority of these products have a long history of use, but non-harmonized trade and differences in regulatory policies between countries impact their value chains and lead to concerns over product efficacy, safety and quality. Veronica officinalis L. (common speedwell), a member of Plantaginaceae family, has a long history of use in European traditional medicine, mainly in central eastern Europe and the Balkans. However, no specified control tests are available either to establish the quality of derived herbal products or for the discrimination of its most common substitute, V. chamaedrys L. (germander speedwell). In this study, we use DNA metabarcoding and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) to authenticate sixteen V. officinalis herbal products and compare the potential of the two approaches to detect substitution, adulteration and the use of unreported constituents. HPLC-MS showed high resolution in detecting phytochemical target compounds, but did not enable detection of specific plant species in the products. DNA metabarcoding detected V. officinalis in only 15% of the products, whereas it detected V. chamaedrys in 62% of the products. The results confirm that DNA metabarcoding can be used to test for the presence of Veronica species, and detect substitution and/or admixture of other Veronica species, as well as simultaneously detect all other species present. Our results confirm that none of the herbal products contained exactly the species listed on the label, and all included substitutes, contaminants or fillers. This study highlights the need for authentication of raw herbals along the value chain of these products. An integrative methodology can assess both the quality of herbal products in terms of target compound concentrations and species composition, as well as admixture and substitution with other chemical compounds and plants.

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Centaurea solstitialis L. (yellow starthistle, Asteraceae) is a Eurasian native plant introduced as an exotic into North and South America, and Australia, where it is regarded as a noxious invasive. Changes in ploidy level have been found to be responsible for numerous plant biological invasions, as they are involved in trait shifts critical to invasive success, like increased growth rate and biomass, longer life-span, or polycarpy. C. solstitialis had been reported to be diploid (2n = 2x = 16 chromosomes), however, actual data are scarce and sometimes contradictory. We determined for the first time the absolute nuclear DNA content by flow cytometry and estimated ploidy level in 52 natural populations of C. solstitialis across its native and non-native ranges, around the world. All the C. solstitialis populations screened were found to be homogeneously diploid (average 2C value of 1.72 pg, SD = +/- 0.06 pg), with no significant variation in DNA content between invasive and non-invasive genotypes. We did not find any meaningful difference among the extensive number of native and non-native C. solstitialis populations sampled around the globe, indicating that the species invasive success is not due to changes in genome size or ploidy level.

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Ramaria largentii Marr & D. E. Stuntz (orange coral mushroom) is a wild edible mushroom whose chemical composition and bioactivity have not been investigated. Herein, we present a study on the phenolic constituents, antioxidant and antigenotoxic effects of a hydromethanolic extract of the fruiting bodies. Total phenolic content, estimated by Folin-Ciocalteu assay, was found to be 42.33 +/- 0.18 mg GAE/g. Protocatechuic and vanillic acids were detected by HPLC-DAD-ESI-MS. The extract showed good free radical scavenging and reducing capacities (EC50 = 64.3 +/- 0.2 and 61.54 +/- 0.46 mu g/mL, respectively). In normal Vero cells, the extract (100, 200 and 300 mu g/mL) showed no genotoxic potential and moreover, almost completely protected DNA against H2O2-induced damage (2.09-7.91% tail DNA) (24 and 48 h pretreatment). Taken together, the results of our study show that Ramaria largentii extract is devoid of genotoxicity and has a remarkable DNA protective activity against H2O2-induced damage. (C) 2017 Elsevier Ltd. All rights reserved.

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Ramaria largentii Marr & D. E. Stuntz (orange coral mushroom) is a wild edible mushroom whose chemical composition and bioactivity have not been investigated. Herein, we present a study on the phenolic constituents, antioxidant and antigenotoxic effects of a hydromethanolic extract of the fruiting bodies. Total phenolic content, estimated by Folin-Ciocalteu assay, was found to be 42.33 ± 0.18 mg GAE/g. Protocatechuic and vanillic acids were detected by HPLC-DAD-ESI-MS. The extract showed good free radical scavenging and reducing capacities (EC50 = 64.3 ± 0.2 and 61.54 ± 0.46 μg/mL, respectively). In normal Vero cells, the extract (100, 200 and 300 μg/mL) showed no genotoxic potential and moreover, almost completely protected DNA against H2O2-induced damage (2.09–7.91% tail DNA) (24 and 48 h pre-treatment). Taken together, the results of our study show that Ramaria largentii extract is devoid of genotoxicity and has a remarkable DNA protective activity against H2O2-induced damage. © 2017 Elsevier Ltd

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Background Rochefortia is a small taxon of woody plants in the Ehretiaceae (Boraginales) exhibiting coriaceous leaves with cystoliths, small whitish flowers and drupaceous fruits containing four pyrenes. It shares the dioecious sex distribution with its sister group Lepidocordia and can be delimited from the latter (and all other Ehretiaceae) by the presence of thorns. Neotropical Rochefortia is distributed over most Caribbean islands, Central America and northern South America. Twenty-eight validly published names (corresponding to twenty-one typified taxa at the species level and below) are available in Rochefortia, but the precise number of species to be accepted has been elusive before this revision. New information In the course of the present revision, 353 herbarium collections, comprising approximately 540 Rochefortia specimens, were entried into a BRAHMS data base providing information about protologues and types and retrospective georeferences if possible. Based on the combination of molecular and morphological data we propose to recognise nine species of Rochefortia, namely R. acanthophora, R. bahamensis, R. barloventensis, R. cubensis, R. cuneata, R. lundellii, R. oblongata, R. spinosa and R. stellata (the remaining nineteen validly published names are synonymised under such names). Morphological description of each species and an identification key are provided.

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The plastid-nucleus located protein WHIRLY1 has been described as an upstream regulator of leaf senescence, binding to the promoter of senescence-associated genes like HvS40. To investigate the impact of WHIRLY1 on drought stress-induced, premature senescence, transgenic barley plants with an RNAi-mediated knockdown of the HvWHIRLY1 gene were grown under normal and drought stress conditions. The course of leaf senescence in these lines was monitored by physiological parameters and studies on the expression of senescence- and drought stress-related genes. Drought treatment accelerated leaf senescence in WT plants, whereas WHIRLY 1 knockdown lines (RNAi-W1) showed a stay-green phenotype. Expression of both senescence-associated and drought stress-responsive genes, was delayed in the transgenic plants. Notably, expression of transcription factors of the WRKY and NAC families, which are known to function in senescence- and stress-related signaling pathways, was affected in plants with impaired accumulation of WHIRLY1, indicating that WHIRLY1 acts as an upstream regulator of drought stress-induced senescence. To reveal the epigenetic indexing of HvS40 at the onset of drought-induced senescence in WT and RNAi-W1 lines, stress-responsive loading with histone modifications of promoter and coding sequences of HvS40 was analyzed by chromatin immunoprecipitation and quantified by qRT-PCR. In the wildtype, the euchromatic mark H3K9ac of the HvS40 gene was low under control conditions and was established in response to drought treatment, indicating the action of epigenetic mechanisms in response to drought stress. However, drought stress caused no significant increase in H3K9ac in plants impaired in accumulation of WHIRLY1. The results show that WHIRLY1 knockdown sets in motion a delay in senescence that involves all aspects of gene expression, including changes in chromatin structure.

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The aim of the present study was to investigate the antigenotoxic and antioxidant activities of the crude hydromethanolic extract from the aerial parts of European Dracocephalum moldavica L. (Moldavian dragonhead). The total phenolic content estimated by the Folin-Ciocalteu assay was 289.55 +/- 2.63 mg of GAE/g of dry extract, and rosmarinic acid was the major polyphenol of Dracocephalum moldavica extract (107.11 +/- 0.83 mg/g of dry extract). In vitro antioxidant assays revelead remarkable scavenging effects against DPPH (EC50 = 23.10 +/- 0.10 mu g/mL), ABTS (EC50 = 8.0 +/- 0.10 mu g/mL) and superoxide anion radicals (EC50= 445.5 +/- 2.3 mu g/mL). The extract showed a high ferrous ion chelating activity (EC50 = 35.70 +/- 0.40 mu g/mL), a considerable reducing capacity, and good hydroxyl radical scavenging properties. Dracocephalum moldavica extract reduced, in a concentration-dependent manner, DNA damage induced by bleomycin in normal human dermal fibroblasts as measured by comet assay and micronucleus test. Exposure of dermal fibroblasts to Dracocephalum moldavica extract (100 mu g/mL) after preincubation with bleomycin (10 mu g/mL) resulted in the most significant antigenotoxic activity. The protective effects may be due to the free radical scavenging activity, iron-chelating properties and the possible intervention on DNA repair processes. (C) 2015 Elsevier B.V. All rights reserved.

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Various phytochemicals, mainly phenolic derivatives, have been screened for their radioprotective properties. This review summarizes the current knowledge in radioprotection using plant bioactive compounds with emphasis on two promising compounds, curcumin and genistein. Free radical scavenging abilities, maintaining cellular antioxidant status, modulation of DNA repair or prevention of DNA damages and anti-inflammatory activity are the main mechanisms involved in radioprotection exerted by phytochemicals. The chemical structure-activity relationship, the limits of existing research as well as the suggestions on further studies including clinical application are also discussed.

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The aim of our ongoing study is to assess the quality of the autochthonous plant material of Arnica montana L. species from the northern area of the Romanian Eastern Carpathians. This is the first report for the phytochemical features for the wild populations in this area. The first qualitative assessments were performed by means of High Performance Thin Layer Chromatography (HPTLC). The High Performance Liquid Chromatography (HPLC) analysis highlighted a total content in sesquiterpene-lactones of 0.86-1.36%, while the total content in flavonoids and phenolic acids was 1.08-1.50%, 0.86-1.27% respectively.

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Arnica montana L. is a medicinal and aromatic plant species with high pharmaceutical importance mainly due to the anti-inflammatory proprieties of helenaline. The unsustainable management and environmental changes lead to the extinction of the species in many habitats, becoming endangered or rare in most European countries. The paper synthesises the researches results on the categories of biotic interactions involved in the distribution and abundance of A. montana species in natural habitats. This addresses the impact of phytophagous and mycorrhizal species on bioproductivity, respectively, pollinators on reproductive success and fitness, with importance in developing viable conservation strategies and optimization of cultivation features.

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Knowing that the species of the Thymus genus present important therapeutic properties, in this paper we aim to analyze the essential oils from five spontaneous populations of Thymus pulegioides L. in order to identify populations with potential therapeutic value. For this purpose, essential oils were extracted by hydro-distillation, using a NeoClevenger type apparatus according to the European Pharmacopoeia standards. The separation and the identification of the components were carried out using GC-MS (gas chromatography coupled with mass spectrometry). Antibacterial testing was performed on two test microorganisms, Streptococcus pyogenes ATCC 19615 and Pseudomonas aeruginosa ATCC 27853, using the diffusion method and the microplate method. With respect to the chemical composition of the essential oils, there were significant qualitative and quantitative differences. It was also noted that all essential oils inhibited the growth and development of the two test microorganisms used.

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The present work reports on the biocompatibility of poly (3-hydroxybutyrate) and poly (3-hydroxybutyrateco-3-hydroxyvalerate) loaded with bacterial cellulose and microcrystalline cellulose via melt processing. Biocompatibility was tested by physico-chemical and in vitro methods. Physico-chemical tests of biocomposites, such as reducing substances, acidity, alkalinity, absorbance by UV/VIS, residue on evaporation were performed on aqueous extract. The cell viability was evaluated by MTT assay. Cell morphology evaluation of cell culture treated with composites was visualized by light microscopy. Also, thermal properties of biocomposites were investigated by DSC analysis. The obtained results have shown good biocompatibility of all biocomposites.

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The present study analyzed the possible anxiolytic and antidepressant proprieties of inhaled basil essential oils extracted from Ocimum sanctum L. (Os) and Ocimum basilicum L. (Ob) in beta-amyloid (1-42) rat model of Alzheimer's disease. The anxiolytic- and antidepressant-like effects of inhaled basil essential oils were studied by means of in vivo (elevated plus-maze and forced swimming tests) approaches. The beta-amyloid (1-42)-treated rats exhibited the following: decrease of the exploratory activity (number of crossing), the percentage of the time spent and the number of entries in the open arm within elevated plus-maze test and increase of the swimming time and decrease of the immobility time within forced swimming test. The chemical composition varied from one sample to another. The main compounds found in both samples were linalool (31 %-Ob, 19 %-Os), camphor, beta-elemene, alpha-bergamotene and bornyl-acetate, estragole (15.57 and 7.59 %, respectively), eugenol (2.64 and 1.39 %, respectively) and 1,8-cineole (3.29 and 3.90 %, respectively). As a result, exposure to basil essential oils significantly improved the behavior of the animals, suggesting anxiolytic- and antidepressant-like effects. Our results suggest that multiple exposures to basil essential oils can be useful as a mean to counteract anxiety and depression in Alzheimer's disease conditions.

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Dioecious and thorny Rochefortia Sw. is a poorly known though distinct element of the Ehretiaceae comprising woody plants restricted to the Caribbean and the adjacent American mainland. The approximately ten species display a great morphological variability and overlapping taxonomic boundaries, which makes it difficult to differentiate them (particularly in the Caribbean region). We investigated the phylogenetic relationships of Rochefortia using DNA sequence data from one nuclear locus (Internal Transcribed Spacer) and three chloroplast DNA loci (rps16, trnL-trnF, trnS-trnG). The monophyly of Rochefortia was confirmed, with a sister group relationship between an American mainland clade and a Caribbean clade. The latter segregates into three, morphologically rather variable lineages, distributed either in the Lesser Antilles or in the eastern Greater Antilles or in the western Greater Antilles. Thus, geographic occurrence rather than morphology is indicative of taxonomic delimitation in Rochefortia.

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Taxonomic diversity of Neotropical Rochefortia is not completely assessed at present. We report the existence of a new species: Rochefortia barloventensis sp. nov., distributed across multiple islands of the Lesser Antilles. We provide a morphological description, a molecular diagnosis and a botanical illustration. Specimens belonging to the new species were previously assigned to Caribbean R. cuneata or to South American R. spinosa because of morphological similarity. Molecular sequence data shows a clear delimitation of the new species from all other species of Rochefortia justifying the recognition of a novel taxon.

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The aim of the present study was to investigate the chemical composition and antioxidant activity of Satureja montana essential oil of Romanian origin (S. montana EO). The essential oil was obtained by hydrodistillation from dried aerial parts of cultivated plants in northeastern Romania. The chemical composition of S. montana EO was analysed by GC and GC-MS. The main compounds were carvacrol (63.40%), para-cymene (10.97%) and gamma-terpinene (3.70%). S. montana EO showed a high reducing capacity (IC50 = 64.35 mu g/mL), and was more effective as DPPH free radical scavenger (IC50 = 243.80 +/- 4.93 mu g/mL) than ABTS(circle+) inhibitor (IC50 = 560.86 +/- 23.15 mu g/mL). S. montana essential oil of Romanian origin belongs to the carvacrol chemotype and represents a suitable agent for potential applicability in the protection of biological matrices from the free radical-mediated oxidative stress, including ionizing radiation-induced oxidative damage.

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Both free radicals and oxygen reactive species have been involved as mediators to cell and tissue injury, inflammation, ischemia, neurodegeneration and dementia. Therefore, a great amount of evidence-based studies tried to explain the mechanisms and structure-relation activity. Most of these studies are related to polyphenols, renowned compounds for their powerful antioxidant properties. Our research aimed to evaluate the antioxidant capacity of the essential oil extracted from Salvia officinalis L. from biologic cultures. We used in vitro and in vivo (the determination of superoxide dismutase activity and acetylcholinesterase inhibitory activity) studies to fully assess its potential. The GC-MS analysis indicated the presence of 45 compounds and the principal components of the essential oil, in addition to thujone, were 1,8-cineol and camphor. The results of in vitro tests indicated that Salviae aetheroleum is a powerful inhibitor against 15-lipoxygenase (IC50 0.064 mu L/mL and cholinesterase (IC50 0.478 mu L/mL) and a good scavenger of free radicals (IC50 10.5 mu L/mL). For the in vivo testing, the essential oil was administered by inhalation to rats with induced Alzheimer disease and brain tissue samples were analysed. Daily exposures for one week to sage essential oil increased antioxidant enzymes activity, suggesting that the main mechanism to prevent neurodegeneration is related to antioxidant properties.

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Pharmacovigilance of herbal medicines relies on the product label information regarding the ingredients and the adherence to good manufacturing practices along the commercialisation chain. Several studies have shown that substitution of plant species occurs in herbal medicines, and this in turn poses a challenge to herbal pharmacovigilance as adverse reactions might be due to adulterated or added ingredients. Authentication of constituents in herbal medicines using analytical chemistry methods can help detect contaminants and toxins, but are often limited or incapable of detecting the source of the contamination. Recent developments in molecular plant identification using DNA sequence data enable accurate identification of plant species from herbal medicines using defined DNA markers. Identification of multiple constituent species from compound herbal medicines using amplicon metabarcoding enables verification of labelled ingredients and detection of substituted, adulterated and added species. DNA barcoding is proving to be a powerful method to assess species composition in herbal medicines and has the potential to be used as a standard method in herbal pharmacovigilance research of adverse reactions to specific products.

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The in vivo phytotoxicity of Romanian caraway (Carum carvi) fruits was assayed by Triticum aestivum L. test. We studied the effects of methanolic caraway fruit extracts upon germination, elongation and accumulation of dry biomass in the plants; in addition, it was investigated the influence of the extracts on mythosis in root tissues. The caraway extracts did not inhibit root and shoot elongation of Triticurn aestivum plants, but enhanced seedling growth, as revealed by the increase of root and shoot length (up to 11.05%) and dry biomass (up to 15.81%). The frequency of chromosomal aberrations observed in root meristems varied slightly around the percentage of the control. The investigated caraway fruit extracts did not have phytotoxic effects but had a slight stimulating effect on germination and growth of Triticum aestivum L.

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Arnica montana is a rare medicinal plant with traditional use in Europe and North America. Modern cultivation technologies (conventional and non-conventional) support the conservation strategies of the species. This paper presents a comparative analysis of plant material obtained in vivo and in vitro (micropropagation, callus culture and submerged culture). For in situ and ex situ samples, the material consisted in aerial and underground parts, while the material from in vitro cultures was represented by plantlets, callus and cell suspensions. For in vitro cultures we used Murashige and Skoog (MS, 1962) basal medium, tissue culture beeing supplemented with benzil-amino-purine (BAP), and callus cultures with BAP, kinetin (6-furfuryl aminopurine) and 2,4-dichlorophenoxyacetic acid. These media were solidified with different amounts of agar. The cell cultures were maintained in flasks with MS liquid medium, placed on a shaker. Phytochemical analysis of absolute methanol and dichloromethane extracts, respectively, were performed by Thin Layer Chromatography and RP-HPLC-UV (Agilent 1200 series system). They revealed the presence of terpenoids, phytosterols and polyphenolic compounds which varied quantitatively and qualitatively in the experimental samples. As a general trend, the maintenance of biosynthetic capacity of the compounds analyzed was observed.

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Veronica officinalis, used in ethno-pharmacy, is a potential source of steroidal glycosides for industrial production. A cosmetic line based on organic extracts of MAP from alpine and subalpine areas, including V. officinalis, is a way to exploit bioresources. As a result of temperature fluctuations and high level of UV radiations in the natural habitat of the species, plants that grow at high altitude have a high content of antioxidant and skin protective compounds. Natural areas in the Romanian Eastern Carpathians, where we identified V. officinalis populations, are under intensifying deforestation, a phenomenon that leads to impairment of essential habitat for species survival. The aim of our study is the use of conventional and non-conventional techniques to obtain V. officinalis both for in situ conservation and for propagation in agro-systems. This requires the supply of vegetal raw material in the context of the reduction of the natural basins and of deforestation in the Romanian Carpathians. Supplementation with herbs is based, in recent practices, on the collection of wild and/or ex situ production in agro-ecosystems. Biological material used to initiate experimental cultures was taken from natural populations in Bistrita Valley. The field cultures were achieved by seeds and bush division, the first method being an advantage from the economic point of view, and the second to maintain uniformity in the selected plant cultures. The biggest production of fresh herba (5100 kg/ha) was achieved by dividing the bushes, in April. In vitro plant regeneration from axillary bud multiplication produced vigorous plantlets after 4-5 weeks, with 85-90% survival rate in field.

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The aim of this study was to assess the antibacterial and antifungal potential of some Romanian medicinal plants, arnica--Arnica montana, wormwood--Artemisia absinthium and nettle--Urtica dioica. In order to perform this antimicrobial screening, we obtained the vegetal extracts and we tested them on a series of Gram-positive and Gram-negative bacteria, and also against two fungal strains. The vegetal extracts showed antimicrobial activity preferentially directed against the planktonic fungal and bacterial growth, while the effect against biofilm formation and development was demonstrated only against S. aureus and C. albicans. Our in vitro assays indicate that the studied plant extracts are a significant source of natural alternatives to antimicrobial therapy, thus avoiding antibiotic therapy, the use of which has become excessive in recent years.

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