Crithmum maritimum L.-Study on the Histochemical Localization of Essential Oil.

Crithmum maritimum L. is a perennial halophyte plant that is a medicinal herb known by people from different cultures since ancient times. However, the therapeutic potential of this halophyte has not been completely investigated, and the scientific data on it are limited. The purpose of the present study was to estimate the chemical composition of the essential oil (EO) obtained from the aerial parts of Crithmum maritimum L. growing wild in Bulgaria, as well as the histolocalization of secretory structures for the synthesis and accumulation of volatile oils. The results obtained on the EO composition of Bulgarian Crithmum maritimum L. provide an opportunity to reveal potential future applications in various fields, such as medicine, pharmacy, agriculture, food, and the cosmetic industry. Gas chromatography with mass spectrometry was performed to assess the chemical profile of the isolated EO. The phenylpropanoid dillapiole was identified as the major compound in the EO, accounting for 34.09% of the total EO. Monoterpene hydrocarbons represented 62.07% of the total oil composition. γ-Terpinene, D-limonene, and ß-pinene were the most abundant monoterpene hydrocarbons in the composition of the EO. In addition, histochemical localization of EO in the stem and leaves of Crithmum maritimum L. was carried out. The secretory structures were located in the cortical region of the stem and in the mesophyll tissues of the leaves in the form of secretory ducts. The performed histochemical analysis confirmed the lipophilic nature of the secretion from the duct cells. This is the first report related to the histolocalization and chemical composition of the EO from Bulgarian Crithmum maritimum L. Furthermore, our data indicate some potential possibilities for the evaluation of the therapeutic activity of the EO obtained from this plant species and outline its future applications as a therapeutic agent. Also, the EO from the studied halophyte plant has prominent potential to be used as a biopesticide, which is an environmentally friendly option compared to standard pesticides.

Chemical Composition and Future Perspectives of Essential Oil Obtained from a Wild Population of Stachys germanica L. Distributed in the Balkan Mountains in Bulgaria.

Stachys germanica L. (Lamiaceae) is a plant associated with a rich history in the traditional medicine of Iran, Turkey, and Serbia. However, researchers have not fully investigated the pharmacological potential of the herb, and scientific data on this plant species are limited. The aim of the current study was to evaluate the chemical composition of the essential oil (EO) obtained from the aerial parts of S. germanica L. growing wild in Bulgaria and to perform a comparative analysis of the chemical composition of EOs obtained from the same plant species from other geographical regions. For the evaluation of the chemical profile of the isolated EO, gas chromatographic analysis with mass spectrometry was performed. The most abundant terpene class was oxygenated monoterpenes, which accounted for 59.30% of the total EO composition. The bicyclic monoterpene camphor, as a compound of this class, was identified as the major constituent in the EO, accounting for 52.96% of the total oil composition. The chemical profile of Bulgarian EO is quite different compared to that of EOs from other regions. It is the only one to contain more than 50% camphor. In addition, EO contains significant amounts of the diterpene geranyl p-cymene (10.49%). This is the first study describing the chemical composition of EO from Bulgarian Stachys germanica L., and our results reveal some future perspectives for the evaluation of the biological activity of EO from the studied plant species as a new therapeutic agent or natural remedy targeting different medical conditions. The EO has a promising potential to be used as a biopesticide and repellent as well, an environmentally friendly and safer alternative to standard pesticides.

A Study of the Chemical Composition, Antioxidant Potential, and Acute Toxicity of Bulgarian Tanacetum vulgare L. Essential Oil.

Common tansy (Tanacetum vulgare L.) is a plant with medicinal properties that has traditionally been used in folk medicine for its anthelmintic, antispasmodic, and choleretic effects, for the treatment of diarrhea and digestive problems, and externally, as an insecticide in veterinary practices. In the current study, we investigated, for the first time, the chemical profile and antioxidant activity of essential oil from a wild population of T. vulgare L. growing in Bulgaria. Common tansy essential oil (EO), which is rich in bicyclic monoterpenes, was obtained using hydrodistillation and characterized by using gas chromatography-mass spectrometry (GC-MS). Thirty-seven compounds were identified in Bulgarian tansy EO. Among the major constituents were oxygenated monoterpenes, including compounds such as camphor (25.24%), trans-chrysantenyl acetate (18.35%), cis-verbenol (10.58%), thujone (6.06%), eucaliptol (5.99%), and α-campholenal (5.98%). The analysis results identified the essential oil from T. vulgare L. grown in the western Rhodope Mountains of Bulgaria as the camphor chemotype. Furthermore, its antioxidant activity was analyzed using the oxygen radical absorbance capacity (ORAC) method and was found to be 605.4 ± 49.3 µmol TE/mL. The essential oil was also tested for single-dose acute toxicity on Wistar rats and was found to be non-toxic by oral administration. The mean lethal dose by intraperitoneal administration was LD50 i.p. = 14.9 g/kg body weight. The results of the conducted study can serve as a basis for the evaluation and subsequent exploration of other pharmacotherapeutic effects of the essential oil obtained from the inflorescences of the Bulgarian species T. vulgare L.

Anti-Adipogenic Activity of Rhaponticum carthamoides and Its Secondary Metabolites.

Besides their common use as an adaptogen, Rhaponticum carthamoides (Willd.) Iljin. rhizome and its root extract (RCE) are also reported to beneficially affect lipid metabolism. The main characteristic secondary metabolites of RCE are phytoecdysteroids. In order to determine an RCE's phytoecdysteroid profile, a novel, sensitive, and robust high-performance thin-layer chromatography (HPTLC) method was developed and validated. Moreover, a comparative analysis was conducted to investigate the effects of RCE and its secondary metabolites on adipogenesis and adipolysis. The evaluation of the anti-adipogenic and lipolytic effects was performed using human Simpson-Golabi-Behmel syndrome cells, where lipid staining and measurement of released glycerol and free fatty acids were employed. The HPTLC method confirmed the presence of 20-hydroxyecdysone (20E), ponasterone A (PA), and turkesterone (TU) in RCE. The observed results revealed that RCE, 20E, and TU significantly reduced lipid accumulation in human adipocytes, demonstrating their anti-adipogenic activity. Moreover, RCE and 20E were found to effectively stimulate basal lipolysis. However, no significant effects were observed with PA and TU applications. Based on our findings, RCE and 20E affect both lipogenesis and lipolysis, while TU only restrains adipogenesis. These results are fundamental for further investigations.

A Study of the Chemical Composition, Acute and Subacute Toxicity of Bulgarian Tanacetum parthenium Essential Oil.

BACKGROUND: Tanacetum parthenium (L.) Sch.Bip. (T. parthenium) is an aromatic perennial plant belonging to the Asteraceae family, also known as feverfew. It is widely distributed in various regions of Europe and other parts of the world. The plant has a rich background in the traditional medicine of many nations and has been used as a remedy for fever, pain, inflammation, asthma, rheumatism, menstrual disorders, etc. Methods: GC-MS analysis was conducted to determine the chemical composition of the isolated essential oil (EO). Using the method proposed by Litchfield and Wilcoxon, the average lethal dose (LD50) of the EO on Wistar rats was determined for two routes of administration: oral (p.o.) and intraperitoneal (i.p.). The subacute toxicity of the EO was also tested by oral administration of a daily dose of 1.0 g/kg body weight (BW) for 28 days. The toxicity of the EO was evaluated by observing and evaluating changes in behavior, body weight, basic hematological and serum biochemical parameters, and histopathological changes of the internal organs. RESULTS: Thirty-seven volatile organic compounds representing 94.58% of the total oil composition were tentatively detected in the obtained T. parthenium EO. The dominant compounds were camphor (45.47%), trans-chrisantenyl acetate (21.65%), camphene (9.48%), and cis-isogeraniol (5.42%). The results showed that the EO was not toxic when administered in acute oral doses. The acute mean lethal dose for intraperitoneal administration was LD50 i.p. = 2.13 g/kg BW. In the subacute study involving administration of an oral dose of EO for 28 days, there were a number of changes in the hematological and serum biochemical parameters of the blood compared with the control group of animals. However, no symptoms of toxicity, changes in the body weight of the rats, death, or pathological changes in the histological indicators of the examined organs-brain, heart, stomach, liver, spleen and kidney-were found. Extrapolating the results obtained from the rat experiments, we can state that the EO is safe for use in doses below 1 g/kgBW for a period not exceeding one month.

Echinophora tenuifolia subsp. sibthorpiana-Study of the Histochemical Localization of Essential Oil.

BACKGROUND: Echinophora tenuifolia L. subsp. sibthorpiana is a perennial, aromatic plant used in traditional folk medicine and cuisine of the Mediterranean and the Middle East. However, scholars have not fully studied the pharmacological potential of the herb, and the scientific data on this plant species are limited. This study aimed to evaluate the chemical composition of the essential oil (EO) obtained from the aerial parts of E. tenuifolia subsp. sibthorpiana growing wild in Bulgaria and to perform histochemical analysis. METHODS: A microscopic histochemical analysis and gas chromatography with mass spectrometry were performed. RESULTS: The histochemical analysis confirmed the presence of terpenes in the stem and leaf of E. tenuifolia subsp. sibthorpiana. The phenylpropanoid methyleugenol was identified as the main compound in the EO, representing 48.13% of the total oil composition. There were also found considerable amounts of monoterpene hydrocarbons, representing 41.68% of the total EO. Alpha-phellandrene, o-cymene, and ß-phellandrene were the most abundant monoterpene hydrocarbons. CONCLUSION: This is the first histochemical analysis performed on E. tenuifolia subsp. sibthorpiana. This is the first report of the EO composition from Bulgarian E. tenuifolia subsp. sibthorpiana, and our results indicate some future possibilities for evaluating of the biological activity of the EO of E. tenuifolia subsp. sibthorpiana and highlight the potential future use of the EO of this plant species. E. tenuifolia L. subsp. sibthorpiana EO possesses a good potential for use as a biopesticide and repellent an environmentally friendly alternative of synthetic pesticides.

Lead content in weight loss food supplements.

INTRODUCTION: The control and prevention of lead exposure are widely discussed topics that seem to be current health concerns for the next decades. Ingestion of lead into the human body can cause toxicity which can be acute or chronic. Possible sources of lead exposure can be, among others, water, food, drugs, and food supplements. The lead content in drugs is a strictly controlled parameter worldwide. The different Pharmacopoeias (European, British, USA) provide information about the limits for lead in every drug substance and also about the tests that should be performed. Regulation of food supplements does not require obligatory analytical control. Often, these products are not monitored to ensure the quality. At the same time, during the last two decades, the global use of dietary supplements has increased manifold.

Dietary Supplements as Source of Unintentional Doping.

Background: The substances used in sport could be divided into two major groups: those banned by the World Anti-Doping Agency and those which are not. The prohibited list is extremely detailed and includes a wide variety of both medicinal and nonmedicinal substances. Professional athletes are exposed to intense physical overload every day. They follow a relevant food regime and take specific dietary supplements, which is essential for the better recovery between trainings and competitions. However, the use of "nonprohibited" dietary supplements (DS) is not always completely safe. One of the risks associated with the use of dietary supplements is the risk of unintended doping-originating from contaminated products. The presence of undeclared compounds in the composition of DS is a serious concern. The aim of this study is to evaluate the risk of unintentional doping. Materials and Methods: Literature search was done through PubMed, Science Direct, Google Scholar, and Web of Science. Studies investigating the presence of undeclared compounds, in dietary supplements, banned by WADA met the inclusion criteria. The last search was conducted in June 2021. The present review is based on a total of 50 studies, which investigated the presence of undeclared compounds in DS. Results: The total number of analyzed DS is 3132, 875 of which were found to contain undeclared substances. Most frequently found undeclared substances are sibutramine and anabolic-androgenic steroids. Conclusion: More than 28% of the analyzed dietary supplements pose a potential risk of unintentional doping. Athletes and their teams need to be aware of the issues associated with the use of DS. They should take great care before inclusion of DS in the supplementation regime.

Higenamine in Plants as a Source of Unintentional Doping.

BACKGROUND: Higenamine is a ß2 agonist of plant origin. The compound has been included in WADA's prohibited list since 2017. Higenamine may be detected in different plants and many food supplements of natural origin. METHODS: Our literature search was conducted through PubMed, Science Direct, Google Scholar, and Web of Science studies investigating the presence of higenamine in plants that are used in traditional folk medicine or included in food supplements. Our study aimed to assess the risk of adverse analytical findings caused by higenamine-containing plants. RESULTS: Based on our literature search, Nelumbo nucifera, Tinospora crispa, Nandina domestica, Gnetum parvifolium, Asarum siebodii,Asarum heterotropoides, Aconitum carmichaelii, and Aristolochia brasiliensis are higenamine-containing plants. Based on data from Eastern folk medicine, these plants can provide numerous health benefits. Professional athletes likely ingest these plants without knowing that they contain higenamine; these herbs are used in treatments for different conditions and various foods/food supplements in addition to folk medicine. CONCLUSION: Athletes and their teams must be aware of the issues associated with the use of plant-based products. They should avoid consuming higenamine-containing plants during and outside of competition periods.

Plant Adaptogens-History and Future Perspectives.

Adaptogens are synthetic compounds (bromantane, levamisole, aphobazole, bemethyl, etc.) or plant extracts that have the ability to enhance the body's stability against physical loads without increasing oxygen consumption. Extracts from Panax ginseng, Eleutherococcus senticosus, Rhaponticum carthamoides, Rhodiola rosea, and Schisandra chinensis are considered to be naturally occurring adaptogens and, in particular, plant adaptogens. The aim of this study is to evaluate the use of plant adaptogens in the past and now, as well as to outline the prospects of their future applications. The use of natural adaptogens by humans has a rich history-they are used in recovery from illness, physical weakness, memory impairment, and other conditions. About 50 years ago, plant adaptogens were first used in professional sports due to their high potential to increase the body's resistance to stress and to improve physical endurance. Although now many people take plant adaptogens, the clinical trials on human are limited. The data from the meta-analysis showed that plant adaptogens could provide a number of benefits in the treatment of chronic fatigue, cognitive impairment, and immune protection. In the future, there is great potential to register medicinal products that contain plant adaptogens for therapeutic purposes.

Comparison between the Biological Active Compounds in Plants with Adaptogenic Properties (Rhaponticum carthamoides, Lepidium meyenii, Eleutherococcus senticosus and Panax ginseng).

BACKGROUND: In the 1960s, research into plant adaptogens began. Plants with adaptogenic properties have rich phytochemical compositions and have been used by humanity since ancient times. However, it is not still clear whether the adaptogenic properties are because of specific compounds or because of the whole plant extracts. The aim of this review is to compare the bioactive compounds in the different parts of these plants. METHODS: The search strategy was based on studies related to the isolation of bioactive compounds from Rhaponticum carthamoides, Lepidium meyenii, Eleutherococcus senticosus, and Panax ginseng. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed. RESULTS: This review includes data from 259 articles. The phytochemicals isolated from Rhaponticum carthamoides, Lepidium meyenii, Eleutherococcus senticosus, and Panax ginseng were described and classified in several categories. CONCLUSIONS: Plant species have always played an important role in drug discovery because their effectiveness is based on the hundreds of years of experience with folk medicine in different nations. In our view, there is great potential in the near future for some of the phytochemicals found in these plants species to become pharmaceutical agents.

Regulatory Requirements for Food Supplements in the European Union and Bulgaria.

Data provided by the European Commission show that public expenditures for food supplements have been constantly increasing over the last decade. The aim of the present study was to analyze the major food supplements regulations in the European Union and in Bulgaria. Relevant publications were searched and found in MEDLINE/PubMed, Scopus Database, Web of Knowledge, and in Internet-based search with predefined keywords. Numerous authors support the opinion that there is a real risk that low quality products may be distributed in countries with poor production control. Bulgarian national legislation have 6 regulations with different legal force for provision of healthy and safe food supplements on the market. Currently, the European Community is concerned about food products, including FS, launched on the market because the citizens require reliable protection of their health. This concern provoked the European Parliament and the Council to undertake a number of legislative initiatives in the field of production and control of food products. It is necessary to set more precise quality and safety criteria in future strategies associated with those products.

Osteoporosis: Therapeutic Options.

The definition of osteoporosis was originally formulated at a conference of the World Health Organization (WHO) in 1993 as 'a systemic skeletal disease characterized by decreased bone mass and altered micro-architecture of bone tissue, leading to enhanced bone fragility and risk of fractures'. Osteoporosis is characterized by low bone mineral density (BMD) and loss of the structural and bio-mechanical properties that are required to maintain bone homeostasis. This review aims to address the currently available options in prevention and treatment of osteoporosis. Management of osteoporosis includes non-pharmacological treatment - diet rich of calcium and vitamin D, healthy lifestyle, proper exercise plan, and pharmacological therapy. Combination of non-pharmacological and pharmacological treatment options have to be considered for prevention of osteoporosis and minimization of the risk of fractures. Given the heterogeneity of osteoporosis syndrome and lack of significant number of comparative studies, the choice of a pharmacological agents should be individualized.