Research Report

Pharmacological Effects of Aromatic Medicinal Plants: Comprehensive Analysis of Active Ingredients and Mechanisms of Action  

Xiangjun Dong
College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China
Author    Correspondence author
Medicinal Plant Research, 2024, Vol. 14, No. 1   doi: 10.5376/mpr.2024.14.0002
Received: 01 Jan., 2024    Accepted: 03 Feb., 2025    Published: 11 Feb., 2024
© 2024 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Dong X.J., 2024, Pharmacological effects of aromatic medicinal plants: comprehensive analysis of active ingredients and mechanisms of action, Medicinal Plant Research, 14(1): 11-30 (doi: 10.5376/mpr.2024.14.0002)

Abstract

This study aims to comprehensively evaluate the pharmacological effects of aromatic medicinal plants, synthesize current knowledge on their therapeutic potential, and identify directions for future research. The study identifies essential oils, alkaloids, and flavonoids as key active ingredients in aromatic medicinal plants, responsible for a range of pharmacological activities, including anti-inflammatory, antimicrobial, antiviral, and antioxidant effects. It was found that extracts from these plants exhibit strong antiproliferative effects against various types of cancer and have the ability to modulate critical biological pathways such as NF-κB and MAPK. Additionally, the study emphasizes the need for more detailed pharmacokinetic and pharmacodynamic research on these plants and underscores their potential as complementary or alternative treatments in modern medicine. Due to their diverse and potent active ingredients, aromatic medicinal plants hold significant potential for medical and therapeutic applications. Ongoing research and collaboration among scientists, healthcare providers, and policymakers are essential to fully harness the benefits of these natural resources. Combining traditional knowledge with modern scientific research can unlock new therapeutic potentials and contribute to more holistic and sustainable medical practices.

Keywords
Aromatic medicinal plants; Pharmacological effects; Active ingredients; Anti-inflammatory; Antimicrobial; Therapeutic potential

Aromatic medicinal plants, often referred to as herbs and spices, have been utilized since antiquity for their unique flavors and therapeutic properties. These plants are characterized by their ability to produce essential oils and other volatile compounds that contribute to their distinctive aromas and pharmacological activities. Common examples include Mentha haplocalyxNardostachys jatamansiSyzygium aromaticumAngelica dahurica, and many others, each with unique properties and applications (Fang et al., 2020; Dong et al., 2021). They are widely used in traditional medicine systems across various cultures for treating a range of ailments, from minor infections to chronic diseases (Kieliszek et al., 2020; Spréa et al., 2020). The essential oils extracted from these plants are employed in numerous industries, including cosmetics, food preservation, and pharmaceuticals, due to their antimicrobial, antioxidant, and anti-inflammatory properties (Fitsiou et al., 2019; Batiha et al., 2020).

 

The historical and cultural significance of aromatic medicinal plants is profound, with many traditional medicine systems, such as Ayurveda, Traditional Chinese Medicine, and African ethnomedicine, relying heavily on these plants for their healing properties (El-Shemy et al., 2017; Kaurinović and Vaštag, 2019; Kaurinović et al., 2021). In modern times, there is a growing interest in pharmacological research to scientifically validate the therapeutic claims associated with these plants. Genomic studies play a crucial role in this context by identifying the specific bioactive compounds and elucidating their mechanisms of action. This not only helps in understanding the pharmacodynamics and pharmacokinetics of these compounds but also aids in the discovery of new drugs and therapeutic agents (Fierascu et al., 2021).

 

The primary objective of this study is to comprehensively evaluate the pharmacological effects of aromatic medicinal plants, with a focus on their active ingredients and mechanisms of action. By identifying and categorizing the active compounds present in aromatic medicinal plants, we aim to analyze their pharmacological effects and therapeutic potential. Additionally, we will investigate the cellular and molecular mechanisms through which these compounds exert their effects and assess the current clinical applications and future therapeutic prospects of these plants. By achieving these goals, we hope to bridge the gap between traditional knowledge and modern scientific research, providing a solid foundation for future studies and applications. This comprehensive evaluation will significantly enhance the scientific understanding of the pharmacological field and plant-based medicine, contributing to the development of new plant-based therapeutic agents.

 

1 Overview of Aromatic Medicinal Plants

1.1 Definition and characteristics

Aromatic medicinal plants are defined as those plants that produce essential oils and other volatile compounds, which are responsible for their distinctive fragrances and therapeutic properties. These plants have been used for centuries in traditional medicine, food preservation, and as natural remedies due to their bioactive compounds (Samarth et al., 2017; Cappai et al., 2020).

 

Aromatic medicinal plants are characterized by their ability to synthesize and store essential oils in specialized cells or glands. These essential oils are complex mixtures of volatile compounds, including terpenes, phenolics, and other secondary metabolites. The chemical properties of these compounds contribute to the plants' pharmacological activities, such as antimicrobial, antioxidant, anti-inflammatory, and anticancer effects (Petrović et al., 2019; Batiha et al., 2020).

 

The primary distinction between aromatic and non-aromatic medicinal plants lies in the presence of essential oils and volatile compounds. Aromatic plants are rich in these compounds, which are responsible for their scent and many of their therapeutic effects. In contrast, non-aromatic medicinal plants may possess therapeutic properties but lack the volatile components that characterize aromatic plants (Cappai et al., 2020; Šarčević-Todosijević et al., 2023). Non-aromatic plants often rely on other types of bioactive compounds, such as alkaloids, flavonoids, and glycosides, for their medicinal effects.

 

1.2 Commonly used aromatic medicinal plants

There is a wide variety of aromatic medicinal plants, with common ones including Chinese mint (Mentha haplocalyx), rose (Rosa rugosa), clove (Syzygium aromaticum), and angelica (Angelica dahurica), and other species (Table 1) (Samarth et al., 2017; Batiha et al., 2020; Cappai et al., 2020). These aromatic medicinal plants are distributed across various regions, including Asia, Africa, and the Mediterranean. They thrive in diverse habitats ranging from temperate to tropical climates, often found in regions with well-drained soils and adequate sunlight (Cappai et al., 2020).

 

Table 1 Commonly used aromatic medicinal plants

 

Aromatic medicinal plants have been integral to traditional medicine systems worldwide. For example, Mentha haplocalyx is used in Chinese medicine, Syzygium aromaticum in Indian Ayurveda, and Nardostachys jatamansi in Tibetan medicine. These plants are employed for their therapeutic properties, including treating digestive disorders, respiratory ailments, and inflammatory conditions (Samarth et al., 2017; Kieliszek et al., 2020). These plants not only hold a significant place in traditional medicine but also have widespread applications in modern pharmacological research.

 

1.3 Economic and agricultural importance

Aromatic medicinal plants hold significant economic value in global markets due to their applications in pharmaceuticals, cosmetics, food, and beverage industries. The demand for natural and eco-friendly products has further boosted their market value (Samarth et al., 2017; Cappai et al., 2020). For instance, the global market for peppermint oil is substantial, driven by its demand in oral care products and therapeutic applications. Similarly, the market for clove oil is robust due to its use in dental care and aromatherapy (Zhao et al., 2022; Yadav et al., 2022). These plants also play a crucial role in agriculture and horticulture, often cultivated for their essential oils and bioactive compounds. They contribute to sustainable agricultural practices by promoting biodiversity and serving as natural pest repellents (Cappai et al., 2020; Šarčević-Todosijević et al., 2023). For example, Citrus reticulata is commonly intercropped with other crops to benefit from its pest-deterrent effects.

 

The cultivation and harvesting of aromatic medicinal plants must consider sustainability and environmental impact. Overharvesting and habitat destruction pose significant threats to some species. Sustainable farming practices, such as organic cultivation and responsible wild harvesting, are crucial to ensure the long-term availability of these valuable plants (Cappai et al., 2020; Šarčević-Todosijević et al., 2023). Moreover, initiatives to conserve plant biodiversity and promote sustainable use are essential to maintain ecological balance and support local economies.

 

By understanding the definition, characteristics, and significance of aromatic medicinal plants, we can appreciate their multifaceted roles in traditional medicine, modern pharmacology, and global economies. Further research into their active ingredients and mechanisms of action will continue to uncover their therapeutic potential and contribute to the development of new medicinal products.

 

2 Active Ingredients of Aromatic Medicinal Plants

2.1 Identification of active compounds

Aromatic medicinal plants are rich in various types of active compounds, including essential oils, alkaloids, and flavonoids. Essential oils are volatile compounds responsible for the distinctive aromas of these plants and often contain terpenes, phenylpropanoids, and other secondary metabolites. They are widely used in industries such as cosmetics, flavoring, and herbal beverages (Samarth et al., 2017; Tasneem et al., 2019). Alkaloids are nitrogen-containing compounds known for their potent pharmacological effects. Flavonoids are polyphenolic compounds that exhibit a range of biological activities, such as antioxidant, anti-inflammatory, and antimicrobial properties (Tasneem et al., 2019).

 

The identification and extraction of active compounds from aromatic medicinal plants involve several techniques. Extraction methods, such as steam distillation, solvent extraction, and supercritical fluid extraction, are employed to isolate essential oils and other bioactive compounds from plant materials (Samarth et al., 2017). Chromatographic methods, such as gas chromatography (GC) and high-performance liquid chromatography (HPLC), are commonly used to separate and identify individual components. Spectroscopic methods, including mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy, provide detailed structural information about the compounds.

 

Recent technological advances have significantly enhanced the identification of active compounds in aromatic medicinal plants. High-resolution mass spectrometry (HRMS) allows for the precise determination of molecular weights and structures of complex compounds. Advances in NMR spectroscopy provide more detailed and accurate structural elucidation. Additionally, the integration of chromatography and spectrometry, such as GC-MS and LC-MS, offers powerful tools for comprehensive analysis. Bioinformatics and cheminformatics tools further assist in the interpretation of complex data, leading to a better understanding of the phytochemical diversity and bioactivity.

 

2.2 Phytochemical composition

The chemical composition of major aromatic medicinal plants varies significantly. For instance, oregano and rosemary are rich in polyphenolics, which contribute to their antimicrobial and antioxidant properties (Tasneem et al., 2019). Chinese mint (Mentha haplocalyx) contains menthol, menthone, and other monoterpenes. Clove (Syzygium aromaticum) is rich in eugenol, eugenyl acetate, and caryophyllene. The composition of these compounds can influence the pharmacological properties and efficacy of the plants. Sage and basil contain a high concentration of essential oils, which are responsible for their distinctive aromas and therapeutic effects (Tasneem et al., 2019).

 

Comparative analysis of phytochemical profiles reveals that different aromatic plants possess unique combinations of bioactive compounds. For example, while oregano is rich in carvacrol and thymol, rosemary contains significant amounts of rosmarinic acid and caffeic acid (Tasneem et al., 2019). A comparative study of essential oils from different species of the genus Mentha can show variations in the levels of menthol, menthone, and other components, which may correlate with their therapeutic uses (Bektašević et al., 2021). Such comparative studies help in understanding the specific health benefits and applications of each plant.

 

Environmental factors such as soil type, climate, and altitude significantly influence the phytochemical composition of aromatic medicinal plants. Variations in these factors can lead to differences in the concentration and efficacy of bioactive compounds (Tasneem et al., 2019). For instance, plants grown in different regions or under different climatic conditions may exhibit variations in essential oil yield and composition. Understanding these influences is crucial for optimizing the cultivation and harvesting of these plants to maximize their medicinal properties.

 

2.3 Bioavailability and metabolism

The absorption and bioavailability of active compounds from aromatic medicinal plants are critical for their therapeutic efficacy. Bioavailability refers to the proportion of a compound that reaches the systemic circulation and is available for biological activity. Factors such as solubility, stability, and permeability influence the absorption of these compounds. Essential oils, for instance, are rapidly absorbed through the skin and mucous membranes, making them effective for topical and inhalation therapies (Čabarkapa et al., 2020). However, the bioavailability of polyphenolics can be limited due to their poor solubility and stability in the gastrointestinal tract (Tasneem et al., 2019). Once absorbed, the active compounds undergo various metabolic pathways and biotransformation processes in the body. These metabolic pathways involve processes such as oxidation, reduction, hydrolysis, and conjugation. Essential oils are primarily metabolized in the liver, where they are converted into more water-soluble forms for excretion (Čabarkapa et al., 2020). Polyphenolics, on the other hand, are metabolized by gut microbiota, which can influence their bioactivity and health benefits (Tasneem et al., 2019).

 

Several factors affect the bioavailability of active compounds from aromatic medicinal plants. Delivery methods, such as encapsulation and nanoemulsion, can enhance the stability and absorption of these compounds (Čabarkapa et al., 2020). Interactions with other dietary components and individual variations in metabolism also play a significant role in determining the bioavailability and therapeutic efficacy of these compounds (Tasneem et al., 2019; Álvarez-Martínez et al., 2021). Certain flavonoids can inhibit the activity of metabolizing enzymes, thereby enhancing the bioavailability of co-administered compounds.

 

3 Pharmacological Effects

3.1 Anti-inflammatory effects

Several studies have demonstrated the anti-inflammatory properties of aromatic medicinal plants. For instance, curcumin from Curcuma longa have shown significant anti-inflammatory and immunomodulatory actions, including inhibitory effects on cellular and humoral immunity, lymphocyte activation, and apoptosis propagation (Tasneem et al., 2019). essential oils from plants such as Chinese mint (Mentha haplocalyx), eucalyptus (Eucalyptus globulus), and clove (Syzygium aromaticum) have shown significant anti-inflammatory effects in various in vitro and in vivo models. These studies often involve assays that measure the inhibition of pro-inflammatory cytokines, such as TNF-α and IL-6, as well as the suppression of inflammatory mediators like nitric oxide (NO) and prostaglandins (Park et al., 2022). Furthermore, essential oils from Thymus algeriensis and Artemisia herba-alba have demonstrated substantial anti-inflammatory activity in vivo (Ouahdani et al., 2021).

 

The anti-inflammatory mechanisms of these plants involve multiple pathways, such as curcumin, menthol, eugenol, and 1,8-cineole exert their effects by modulating key signaling pathways involved in inflammation. Curcumin modulate inflammation by affecting signal transduction networks, transcription factors, and the complement system (Tasneem et al., 2019). Park et al. (2022) alleviated neuroinflammation by modulating the CREB/Nrf2/HO-1 pathway, particularly under conditions induced by lipopolysaccharide (LPS). The experimental results showed that menthol (MH) extract inhibited the production of pro-inflammatory enzymes and mediators (such as nitric oxide, tumor necrosis factor-α, and interleukin-6) and reduced the generation of reactive oxygen species (ROS). Additionally, MH inhibited the NF-κB pathway, which plays a crucial role in regulating inflammatory responses (Figure 1). Eugenol has been shown to suppress the expression of COX-2, an enzyme responsible for the production of pro-inflammatory prostaglandins. Additionally, 1,8-cineole modulates the MAPK pathway, leading to a reduction in the production of inflammatory cytokines (Park et al., 2022). The essential oils from Thymus algeriensis and Artemisia herba-alba inhibit edema development induced by carrageenan, indicating their role in modulating inflammatory responses (Ouahdani et al., 2021).

 

Figure 1 Effects of MH on the (A) translocation, (B) cytosol, nuclear protein expression, phosphorylation of NF-κB p65, and phosphorylation and degradation of IκBα (Adopted from Park et al., 2022)

Image caption: figure demonstrates the inhibitory effect of MH on the NF-κB signaling pathway in LPS-induced BV2 cells. Figure 1A shows that LPS stimulation significantly increased the fluorescence intensity of NF-κB p65 in the nucleus, and this effect was reduced in a dose-dependent manner in the presence of MH, indicating that MH inhibited the nuclear translocation of NF-κB p65. Figure 1B shows that LPS treatment increased the phosphorylation and degradation of IκBα, activating NF-κB p65, while MH pretreatment reduced these changes. This further suggests that MH alleviates neuroinflammatory responses by inhibiting the degradation of IκBα and the phosphorylation of NF-κB p65 (Adapted from Park et al., 2022)

 

The anti-inflammatory properties of these plants suggest their potential use in treating chronic inflammatory diseases such as rheumatoid arthritis and atherosclerosis. For example, the combination of Syzygium aromaticum and Rosmarinus officinalis has shown synergistic anti-inflammatory effects, which could be beneficial in pain management and inflammation-related conditions (Déciga-Campos et al., 2021). These findings support the integration of these plants into therapeutic strategies for managing inflammation.

 

3.2 Antimicrobial and antiviral effects

Aromatic medicinal plants have also been shown to possess antimicrobial and antiviral properties. Essential oils from Syzygium aromaticum and thyme (Thymus vulgaris) have shown broad-spectrum antimicrobial activity against bacteria, fungi, and viruses. Studies have demonstrated the effectiveness of these oils against pathogenic microorganisms, including antibiotic-resistant strains such as MRSA (Methicillin-resistant Staphylococcus aureus) and various strains of influenza viruses (García-Oliveira et al., 2022). Additionally, Piper species have demonstrated strong antibacterial and antifungal activities against human pathogens (Salehi et al., 2019).

 

The antimicrobial effects of these plants are primarily attributed to their high content of phenolic compounds and essential oils, such as eugenol, terpinen-4-ol, and thymol. These compounds disrupt microbial cell membranes, inhibit enzyme activity, and interfere with microbial DNA synthesis (Salehi et al., 2019; García-Oliveira et al., 2022). The essential oils from Piper species, for example, contain diverse secondary metabolites that contribute to their broad-spectrum antimicrobial activity (Salehi et al., 2019). Thymol and carvacrol, found in thyme oil, have been shown to interfere with bacterial quorum sensing, thereby inhibiting biofilm formation and microbial communication. Additionally, these compounds can inhibit viral replication by interfering with viral envelope proteins and enzymes essential for viral entry and replication.

 

The antimicrobial properties of these plants make them valuable in the development of natural preservatives for food and pharmaceuticals. They also hold potential as alternative treatments for infections, particularly in the face of increasing antibiotic resistance. The use of Piper species as natural antioxidants and antimicrobial agents in food preservation highlights their practical applications in ensuring food safety and extending shelf life (Salehi et al., 2019).

 

3.3 Antioxidant effects

Aromatic medicinal plants are rich in antioxidants, which have been extensively studied for their ability to neutralize free radicals and prevent oxidative stress. For instance, the essential oils from Thymus algeriensis and Artemisia herba-alba have shown significant free radical scavenging activity and iron reduction capacity (Ouahdani et al., 2021). Similarly, Essential oils from rosemary (Rosmarinus officinalis), lavender (Lavandula angustifolia), and lemon balm (Melissa officinalis) have demonstrated significant antioxidant activity in various assays, such as DPPH, ABTS, and FRAP assays (Koleilat et al., 2017; Gayoso et al., 2018). These studies highlight the capacity of these oils to scavenge free radicals and reduce lipid peroxidation.

 

The antioxidant effects of these plants are mainly due to their ability to neutralize free radicals and reduce oxidative stress. Phenolic compounds and flavonoids found in these plants donate hydrogen atoms or electrons to free radicals, stabilizing them and preventing cellular damage (Hamid et al., 2020; Ouahdani et al., 2021). Such as rosmarinic acid, carnosic acid, and linalool act by donating hydrogen atoms or electrons to neutralize free radicals, thereby preventing cellular damage. Rosmarinic acid, found in rosemary and lemon balm, has been shown to enhance the activity of endogenous antioxidant enzymes, such as superoxide dismutase (SOD) and catalase (Song et al., 2019). Carnosic acid, another potent antioxidant in rosemary, protects cell membranes from oxidative damage by stabilizing lipid structures.

 

The antioxidant properties of aromatic medicinal plants suggest their potential use in preventing and managing oxidative stress-related diseases such as cardiovascular diseases and cancer. Rosemary and lavender oils are incorporated into skincare products for their antioxidant benefits, which help protect the skin from UV damage and premature aging. The potential benefits include improved health outcomes, reduced oxidative damage, and enhanced overall well-being.

 

4 Mechanisms of Action

4.1 Molecular targets

The active ingredients in aromatic medicinal plants interact with specific cellular and molecular targets to exert their pharmacological effects. These targets include receptors, enzymes, ion channels, and signaling molecules. For instance, menthol from peppermint interacts with transient receptor potential (TRP) channels, specifically TRPM8, which is involved in the sensation of cold and pain modulation (Liu et al., 2020).

 

Active compounds from these plants influence various biological pathways. For example, research indicates that eugenol can alleviate inflammatory responses by inhibiting the nuclear factor-kappa B (NF-κB) signaling pathway (Wei et al., 2018). This plays a crucial role in regulating immune and inflammatory responses, capable of reducing the expression of inflammatory factors such as tumor necrosis factor-alpha (TNFα) and interleukin-6 (IL-6), thereby exerting anti-inflammatory effects. Similarly, rosmarinic acid found in rosemary and lemon balm modulates the MAPK pathway, which affects cell proliferation, differentiation, and apoptosis (Jang et al., 2018). These interactions highlight the broad therapeutic potential of aromatic medicinal plants by targeting multiple pathways involved in disease processes.

 

Key molecular targets of active compounds in aromatic medicinal plants include TRPM8 (Transient Receptor Potential Melastatin 8), COX-2, MAPK, and GABA (Gamma-Aminobutyric Acid) receptors. TRPM8 (Transient Receptor Potential Melastatin 8), activated by menthol, leading to analgesic effects (Liu et al., 2020); COX-2 (Cyclooxygenase-2), inhibited by eugenol, reducing inflammation (Ren et al., 2020); MAPK (Mitogen-Activated Protein Kinase), modulated by rosmarinic acid, influencing cell survival and inflammation; and GABA (Gamma-Aminobutyric Acid) receptors, interacted with by linalool from lavender, promoting anxiolytic and sedative effects (Taylor et al., 2020).

 

4.2 Pharmacokinetics and pharmacodynamics

The pharmacokinetics of active ingredients from aromatic medicinal plants involve their absorption, distribution, metabolism, and excretion (ADME). Essential oils are typically absorbed through the gastrointestinal tract, skin, or respiratory system. Once absorbed, these compounds are distributed throughout the body, often binding to plasma proteins. Metabolism primarily occurs in the liver, where enzymes such as cytochrome P450 oxidize, reduce, and conjugate these compounds to enhance their excretion (Xiong et al., 2019). Finally, the metabolites are excreted via urine, feces, or exhalation.

 

The pharmacodynamics of these active ingredients are characterized by dose-response relationships, where the effect of the compound increases with its concentration up to a certain point. Understanding these relationships helps in determining the therapeutic windows, the range of doses that elicit therapeutic effects without causing significant adverse effects. For example, Understanding the relationship between dose and response allows for the calculation of the dose that produces 50% of the maximum effect (ED50), which is crucial for rational drug dosage adjustments. While the dose-response curve obtained from in vitro experiments can provide scientific evidence for the safety and efficacy of a drug, dosage adjustments in actual clinical practice need to take into account individual patient differences (Warren, 2019).

 

Several factors influence the pharmacokinetics and pharmacodynamics of active compounds from aromatic medicinal plants, including delivery methods, chemical structure, and interactions with other compounds. Delivery methods such as oral, topical, and inhalation routes affect the absorption and bioavailability. The chemical structure, including lipophilicity and molecular size, influences distribution and membrane permeability. Additionally, interactions with other herbs or drugs can alter metabolism and efficacy.

 

4.3 Synergistic and antagonistic interactions

Synergistic interactions occur when the combined effect of multiple compounds exceeds the sum of their individual effects. This is often seen in essential oils, where compounds like menthol and eucalyptus oil work together to enhance anti-inflammatory and analgesic effects. Synergism can enhance therapeutic efficacy, allowing for lower doses and reducing the risk of adverse effects. Antagonistic interactions occur when one compound reduces or negates the effect of another. For example, the sedative effects of linalool might be diminished if combined with a stimulant compound (Yunusoğlu, 2021). Additionally, eugenol in clove oil has been shown to enhance the efficacy of certain antibiotics against resistant bacterial strains, highlighting a beneficial synergistic effect. Understanding these interactions is crucial for developing effective and safe herbal formulations.

 

5 Clinical Applications and Therapeutic Potential

5.1 Current clinical uses

Aromatic medicinal plants have been utilized for a variety of therapeutic applications due to their rich content of bioactive compounds. These plants are known for their antimicrobial, antifungal, anti-inflammatory, and antioxidant properties, which have been harnessed in traditional and modern medicine (Vallejo et al., 2017; Samarth et al., 2017; Kieliszek et al., 2020). For instance, essential oils from plants like myrrh (Commiphora myrrha (Nees) Engl.), ginger (Zingiber officinale), and turmeric (Curcuma longa) have shown promising biological activities that can protect against or alleviate various ailments (Kieliszek et al., 2020; Zhang et al., 2023). Additionally, clove (Syzygium aromaticum) has been used for its analgesic, antioxidant, anticancer, antiseptic, antidepressant, antispasmodic, anti-inflammatory, antiviral, antifungal, and antibacterial properties (Batiha et al., 2020).

 

Several clinical trials and case studies have been conducted to evaluate the effectiveness of aromatic medicinal plants. For example, essential oils and extracts from Greek aromatic plants have been studied for their antiproliferative potential against different types of cancer, showing promising results in inhibiting cancer cell growth (Fitsiou and Pappa, 2019). Similarly, the radioprotective activity of certain aromatic plants has been documented, highlighting their potential in protecting against radiation-induced damage (Samarth et al., 2017). Moreover, the use of essential oils and volatile compounds in Alzheimer's disease therapy has been explored, with some studies suggesting their potential in improving cognitive functions (Maggio et al., 2016).

 

The effectiveness of aromatic medicinal plants in clinical settings has been supported by various studies. For instance, plant-derived compounds that modulate oxidative stress have shown potential in cancer therapeutics by inducing cytotoxicity and apoptosis in cancer cells (Vallejo et al., 2017). Additionally, the integration of these plants into the diet has been encouraged to boost overall health, supported by scientific evidence (Kieliszek et al., 2020). Patient outcomes have generally been positive, with many studies reporting significant improvements in symptoms and overall health status (Fitsiou and Pappa, 2019; Kieliszek et al., 2020; Batiha et al., 2020).

 

5.2 Potential for new therapies

Emerging research continues to explore the therapeutic potential of aromatic medicinal plants. Recent studies have focused on the bioactive compounds in these plants and their mechanisms of action, which could lead to the development of new therapies (Vallejo et al., 2017; Krishnaprabu et al., 2020). For example, the antioxidant and anti-inflammatory properties of polyphenols in medicinal plants are being investigated for their potential health benefits (Šarčević-Todosijević et al., 2023). Additionally, the discovery of new bioactive molecules through phytochemical screening has opened up possibilities for novel drug development (Krishnaprabu et al., 2020).

 

Aromatic medicinal plants hold potential for new therapeutic areas beyond their traditional uses. For instance, their radioprotective properties could be harnessed in cancer treatment to protect healthy cells from radiation damage (Samarth et al., 2017). Furthermore, the neuroprotective effects of essential oils and volatile compounds are being explored for the treatment of neurodegenerative diseases like Alzheimer's (Maggio et al., 2016). The antimicrobial and antiviral properties of these plants also present opportunities for developing new treatments for infectious diseases (Batiha et al., 2020).

 

While the therapeutic potential of aromatic medicinal plants is promising, there are several challenges in developing new therapies. One major challenge is the need for more clinical trials to establish the efficacy and safety of these plants in routine healthcare (Maggio et al., 2016). Additionally, the variability in the chemical composition of plant extracts can affect their pharmacological activity, necessitating standardization and quality control (Fitsiou and Pappa, 2019). However, these challenges also present opportunities for further research and innovation in the field of natural product-based drug development (Krishnaprabu et al., 2020; Šarčević-Todosijević et al., 2023).

 

5.3 Integration into conventional medicine

The integration of aromatic medicinal plants with conventional treatments is an area of growing interest. These plants can serve as adjuvants to enhance the efficacy of synthetic drugs or reduce their side effects (Kieliszek et al., 2020). For example, combining plant-derived antioxidants with conventional cancer therapies could improve treatment outcomes by protecting healthy cells from oxidative damage (Vallejo et al., 2017). Additionally, the use of essential oils in aromatherapy has been shown to complement conventional treatments for various conditions, including stress and anxiety (Maggio et al., 2016).

 

Aromatic medicinal plants are widely used in complementary and alternative medicine (CAM) approaches. Their use is based on ethnobotanical evidence and cultural acceptance, making them a popular choice for chronic treatments. CAM approaches often involve the use of whole plant extracts or essential oils, which can provide a synergistic effect due to the presence of multiple bioactive compounds (Kieliszek et al., 2020). These approaches are particularly valuable in regions where access to conventional medicine is limited.

 

Despite their potential, there are several barriers to the integration of aromatic medicinal plants into conventional medicine. These include a lack of standardized protocols for their use, limited clinical evidence, and regulatory challenges (Maggio et al., 2016). To overcome these barriers, more rigorous clinical trials are needed to validate the therapeutic claims of these plants. Additionally, establishing guidelines for the standardization and quality control of plant extracts can help ensure their safety and efficacy (Fitsiou and Pappa, 2019). Collaboration between researchers, healthcare providers, and regulatory agencies is essential to facilitate the integration of these plants into mainstream medicine (Krishnaprabu et al., 2020; Šarčević-Todosijević et al., 2023).

 

6 Safety and Toxicology

6.1 Toxicological profiles

Aromatic medicinal plants have been extensively studied for their safety profiles in both preclinical and clinical settings. Preclinical studies typically involve in vitro and in vivo experiments to evaluate the toxicological effects of these plants' active compounds. For instance, studies on essential oils such as menthol, eugenol, and linalool have shown low toxicity levels when used within recommended doses (Ribeiro-Silva et al., 2022; Wu et al., 2023). Clinical trials have further validated these findings, demonstrating that these compounds are generally safe for humans when used appropriately. Studies have shown that linalool, when used in combination, can significantly enhance its antibacterial effects while maintaining low toxicity levels (Silva et al., 2021). This makes it potentially valuable for applications in food preservation and the medical field. However, it is essential to consider the specific context and formulation, as some compounds may exhibit toxic effects under certain conditions.

 

While aromatic medicinal plants are generally considered safe, they can cause side effects and have contraindications. Common side effects include skin irritation, allergic reactions, and gastrointestinal disturbances. For example, tea tree oil may cause contact dermatitis in sensitive individuals. A study reported two cases where patients developed eczema-like lesions after using products containing tea tree oil, one of whom was a 4-year-old girl (Martínez Campayo et al., 2020). High doses of peppermint oil can cause heartburn or gastroesophageal reflux, and it should be used with caution during pregnancy, breastfeeding, and certain medical conditions. Sanders (2023) found that a postpartum woman developed a systemic rash after taking capsules containing peppermint oil, indicating that peppermint oil may trigger allergic reactions in some cases. Additionally, rosemary oil should be used with caution in patients with epilepsy due to its potential neurostimulatory effects (Liu et al., 2020).

 

The toxicological effects of aromatic medicinal plants are often dose-dependent. At recommended doses, these plants are usually safe and well-tolerated. However, excessive intake can lead to adverse effects. For example, high doses of eugenol from clove oil can cause liver toxicity, and excessive consumption of menthol can result in central nervous system depression (Ribeiro-Silva et al., 2022). It is crucial to adhere to established dosing guidelines to avoid such toxic effects and ensure safe use.

 

6.2 Regulatory considerations

Regulations and guidelines for the use of aromatic medicinal plants vary across regions. In the United States, the Food and Drug Administration (FDA) regulates these plants under the Dietary Supplement Health and Education Act (DSHEA) (Sarma et al., 2021). Essential oils and plant extracts are categorized as dietary supplements, and manufacturers must ensure their safety and efficacy (Shipkowski et al., 2018). In the European Union, the European Medicines Agency (EMA) provides guidelines for the use of herbal medicinal products, requiring evidence of safety and therapeutic efficacy (Qu et al., 2018; Knoess and Wiesner, 2019).

 

Regulatory challenges include the lack of standardized quality control measures and variability in plant composition. Differences in cultivation, harvesting, and processing methods can affect the consistency and potency of the final product. Recent updates in regulations aim to address these issues by implementing stricter quality control standards and requiring more rigorous clinical evidence. For example, the EMA has introduced new guidelines for the standardization of herbal medicinal products to ensure consistent quality and safety (Knöss, 2018).

 

Regulatory frameworks for aromatic medicinal plants differ globally. In the United States, the FDA's approach focuses on post-market surveillance and consumer safety, while the EMA in the European Union emphasizes pre-market evaluation and approval (Qu et al., 2018; Sarma et al., 2021). In Asia, countries like China and India have their traditional medicine systems integrated with modern regulatory practices. China's State Administration for Market Regulation (SAMR) and India's Ministry of AYUSH regulate the use of herbal medicines, ensuring they meet safety and efficacy standards (Seethapathy et al., 2019; Nirmal et al., 2022). Comparing these frameworks highlights the need for international harmonization to facilitate global trade and ensure consumer safety.

 

6.3 Risk assessment and management

Risk assessment of aromatic medicinal plants involves identifying potential hazards, evaluating exposure levels, and determining the likelihood of adverse effects. Methods include toxicological testing, clinical trials, and post-market surveillance. Toxicological testing in preclinical studies assesses acute, subacute, and chronic toxicity (Pande et al., 2018; Sorokina et al., 2019). Clinical trials provide data on safety and efficacy in humans, while post-market surveillance monitors adverse events and ensures ongoing safety.

 

Effective risk management strategies involve implementing safety guidelines, educating consumers, and monitoring product use. Safety guidelines include recommended dosages, contraindications, and potential side effects. Educating consumers about the proper use of these plants and their products is crucial for preventing misuse and adverse effects. Monitoring product use through adverse event reporting systems helps identify emerging safety concerns and allows for timely intervention.

 

Case studies highlight the practical application of risk assessment and management strategies. The safety evaluation of tea tree oil involved extensive preclinical testing and clinical trials to assess its dermatological effects (Martínez Campayo et al., 2020). Risk management strategies included labeling requirements to inform consumers about potential allergic reactions. Another case study on the use of eucalyptus oil in respiratory therapies demonstrated the importance of dose regulation and monitoring for adverse effects, leading to the development of standardized dosing guidelines to ensure safe use.

 

7 Case Study: Syzygium aromaticum

7.1 Overview of Syzygium aromaticum

Syzygium aromaticum, commonly known as clove, is a highly aromatic plant native to the Maluku Islands in Indonesia. It is now also cultivated in other tropical regions such as China, Madagascar, India, Sri Lanka (Chniguir et al., 2019). The plant is characterized by its fragrant flower buds, which are harvested and dried to produce the spice known as clove. Clove trees can grow up to 20 meters tall and have large, glossy leaves and clusters of small, red flowers. The plant is known for its rich content of essential oils and bioactive compounds, which contribute to its diverse pharmacological properties (Chniguir et al., 2019; Xue et al., 2022).

 

Clove has a long history of use in traditional medicine and culinary practices. In traditional Chinese medicine (TCM) and Ayurveda, clove has been used to treat digestive issues, toothaches, and respiratory conditions. The spice was highly valued in ancient trade routes and was a significant part of early global commerce. Historically, clove was also used as a preservative due to its antimicrobial properties, and it played a role in various cultural rituals and traditional health practices.

 

Economically, clove is a valuable commodity in the global spice market (Xue et al., 2022). It is extensively used in culinary applications, particularly in Asian, African, and Middle Eastern cuisines. Clove oil, extracted from the buds, leaves, and stems, is used in pharmaceuticals, dentistry, and aromatherapy. Culturally, clove has been associated with its strong aroma and flavor, symbolizing warmth and festivity in various traditions. Its use in traditional medicine underscores its cultural significance in promoting health and well-being.

 

7.2 Active ingredients in Syzygium aromaticum

The primary active compound in clove is eugenol, which constitutes up to 85% of clove oil. Eugenol is known for its analgesic, antiseptic, and anti-inflammatory properties (Beltrán-Villalobos et al., 2017; Xue et al., 2022). Other significant compounds include isoeugenol, eugenol acetate, β-caryophyllene, and α-humulene. These compounds contribute to the broad pharmacological profile of clove, enhancing its therapeutic applications.

 

Clove oil is typically extracted through steam distillation, which effectively isolates the essential oils from the plant material (Pires et al., 2019). Identification and quantification of the active compounds are performed using techniques such as gas chromatography (GC) and high-performance liquid chromatography (HPLC), coupled with mass spectrometry (MS). Rakhmawati et al. (2022) conducted a chemical analysis of locally produced clove bud oil using gas chromatography-mass spectrometry (GC-MS), identifying 10 major components, including eugenol, trans-caryophyllene, and eugenyl acetate. They also confirmed that the oil met the SNI standards. In another study, total eugenol, eugenol acetate and β-stigmasterol were accurately quantified in clove oil using Fourier transform infrared spectroscopy (ATR-FTIR) and gas chromatography-mass spectrometry (GC-MS) techniques (Tarhan, 2021). The results showed that these techniques can be used for accurate and simultaneous quantification of major compounds in clove oil. 

 

7.3 Pharmacological effects and mechanisms of Syzygium aromaticum

Clove (Syzygium aromaticum) exhibits a wide range of pharmacological activities, including antibacterial, analgesic, anti-inflammatory, and neuroprotective effects (Chniguir et al., 2019; Déciga-Campos et al., 2021; Teles et al., 2021; Benmakhlouf et al., 2022). These properties make it effective in treating infections, pain, inflammation, and oxidative stress-related conditions.

 

Teles et al. (2021) analyzed the chemical composition of clove (Syzygium aromaticum) essential oil and its antibacterial, antioxidant, and anti-trypanosomal activities. The study found that the main component of clove essential oil is eugenol (53.23%), which showed a stronger inhibitory effect against Staphylococcus aureus with a minimum inhibitory concentration of 50 µg/mL. The essential oil also demonstrated significant antioxidant activity and inhibitory effects on Trypanosoma cruzi, the causative agent of Chagas disease, with low selective toxicity to cells, indicating its potential as a novel alternative therapy for Chagas disease (Figure 2). The essential oil and extracts of clove exhibited remarkable antimicrobial properties against various bacterial and fungal strains, making it a potential candidate for treating infections.

 

Figure 2 BALB/c peritoneal macrophages infected with Trypanosoma cruzi and treated with Syzygium aromaticum essential oil or eugenol for 24 hours (Adopted from Teles et al., 2021)

Image caption: The figure shows the infection parameters of BALB/c peritoneal macrophages infected with Trypanosoma cruzi after treatment with different concentrations of clove (Syzygium aromaticum) essential oil and eugenol. Figure 2(a) demonstrates that essential oil at concentrations of 200 µg/mL and 100 µg/mL significantly reduced the number of amastigotes (A non-flagellated form of Trypanosoma cruzi) per 100 cells (p < 0.0068 and p < 0.0460, respectively). Figures 2(b) and 2(d) indicate that 200 µg/mL of eugenol also significantly reduced the number of amastigotes per 100 cells (< 0.0095) and the number of amastigotes per infected cell (< 0.0112). The microscopic images Figure 2(e) show a reduction in the intracellular amastigotes after treatment. These results suggest that clove essential oil and eugenol have significant potential as anti-trypanosomal agents (Adapted from Teles et al., 2021)

 

Chniguir et al. (2019) reported that the aqueous extract of Syzygium aromaticum (SAAE) exhibits significant anti-inflammatory effects by inhibiting the activity of myeloperoxidase (MPO) and metalloproteinases (MMP-2 and MMP-9). The study showed that SAAE effectively inhibits the production of reactive oxygen species (ROS) by neutrophils in vitro and alleviates lipopolysaccharide (LPS)-induced lung inflammation in mice in vivo. It reduces the protein concentration and total cell count in bronchoalveolar lavage fluid. These findings suggest that SAAE has potential value in the development of anti-inflammatory drugs.

 

The study analyzed the analgesic and anti-inflammatory effects of the combination of Syzygium aromaticum (clove) and Rosmarinus officinalis (rosemary) when applied locally in rats (Déciga-Campos et al., 2021). The results showed that both herbs exhibited significant concentration-dependent analgesic and anti-inflammatory effects when used individually. Through isobolographic analysis (Figure 3), it was determined that a 1:1 combination of the two herbs resulted in synergistic effects, with the actual efficacy of the combination being significantly higher than the theoretical value. This suggests that herbal combinations used in traditional medicine may indeed have enhanced therapeutic effects. Furthermore, the study emphasizes the need for pharmacological research into herbal interactions to ensure their efficacy and safety. This finding provides scientific evidence for the application of herbs in pain treatment, supporting their potential in both traditional and modern medicine.

 

Figure 3 Isobolographic analysis of the antinociceptive (a) and anti-inflammatory (b) effect of S aromaticum and R. officinalis in combination (1 :1 ratio) (Adopted from Déciga-Campos et al., 2021)

Image caption: Figure 3 presents the equivalent graph analysis of the combination of rosemary (Rosmarinus officinalis) and clove (Syzygium aromaticum) in analgesic and anti-inflammatory tests. In terms of analgesic effect (Figure (a)), the theoretical value Zadd was 24.8±1.3 μg/paw, while the actual combination value Zmix was 0.45±0.1 μg/paw, indicating a significant synergistic effect. In terms of anti-inflammatory effect (b), the theoretical value Zadd was 14.4±2.2 μg/paw, while the actual combination value Zmix was 5.2±0.6 μg/paw, also showing a significant synergistic effect. These results suggest that the combination of rosemary and clove exhibits a synergistic effect in alleviating pain and inflammation, enhancing the efficacy of both (Adapted from Déciga-Campos et al., 2021)

 

Additionally, Aljarari (2023) explored the neuroprotective effects of clove on aluminum chloride (AlCl3)-induced Alzheimer's disease (AD) in rats. The results showed that clove extract has significant antioxidant properties and can improve cognitive function in AD rats. The study found that clove exhibited neuroprotective effects in the Alzheimer's disease model by improving cognitive function, enhancing acetylcholinesterase levels, and regulating oxidative stress in the brain.

 

7.4 Safety and risk management of clove

When used appropriately, clove is generally safe. However, high doses of eugenol, the main active component, can lead to adverse reactions such as hepatotoxicity and gastrointestinal irritation. Adhering to the recommended dosage is crucial to avoid these toxic effects and ensure safe use. Additionally, individuals allergic to clove or other members of the Myrtaceae family should avoid using it (Ugagu et al., 2021).

 

Effective risk management involves conducting comprehensive risk assessments and following safety guidelines. This includes using standardized extracts, adhering to recommended dosages, and educating consumers about potential risks and proper usage. Monitoring adverse reactions and adjusting treatment plans as necessary are also important strategies to ensure safe and effective use.

 

Clove holds significant therapeutic potential in various fields, including anti-inflammatory, analgesic, antidepressant, and antibacterial applications. To fully harness its medicinal properties and ensure safe use, further research and clinical trials are necessary.

 

8 Challenges and Future Directions

8.1 Research gaps

Despite the extensive research on aromatic medicinal plants, significant gaps remain in our understanding of their pharmacological effects. For instance, while many studies have identified the presence of bioactive compounds, the specific mechanisms through which these compounds exert their effects are often not well understood (Petrović et al., 2019; Saleh-E-In and Choi, 2021; Mesmar et al., 2022). Additionally, there is a lack of comprehensive studies linking traditional uses of these plants with their pharmacological activities, which hinders the development of standardized therapeutic applications (Saleh-E-In and Choi, 2021; Šarčević-Todosijević et al., 2023).

 

Further research is needed to isolate and identify more active compounds from aromatic medicinal plants and to establish the links between these compounds and their pharmacological activities (Saleh-E-In and Choi, 2021; Mesmar et al., 2022). Studies focusing on the structure-activity relationship and in vivo mechanistic studies are particularly important to validate the therapeutic potential of these plants (Ebrahimi et al., 2020; Saleh-E-In and Choi, 2021). Moreover, there is a need for pharmacokinetic evaluation tests to validate the bioavailability and safety of these compounds.

 

To address these gaps, a multidisciplinary approach involving ethnobotany, pharmacology, and advanced analytical techniques should be employed. Collaborative efforts between researchers and traditional medicine practitioners can provide valuable insights into the traditional uses of these plants and guide pharmacological studies (Saleh-E-In and Choi, 2021; Šarčević-Todosijević et al., 2023). Additionally, the use of advanced technologies such as omics techniques can help in understanding the complex interactions between the bioactive compounds and their targets (Christaki et al., 2020).

 

8.2 Technological and methodological advances

Recent innovations in research methodologies, such as the development of robust LC-MS/MS methods, have significantly improved the screening and quantification of phytochemicals in medicinal and aromatic plants (Yılmaz et al., 2020). These advancements allow for more accurate and comprehensive analysis of plant extracts, facilitating the identification of bioactive compounds and their pharmacological properties (Yılmaz et al., 2020). The use of advanced analytical techniques, including liquid chromatography-tandem mass spectrometry (LC-MS/MS) and other high-throughput screening methods, has enhanced the reliability and reproducibility of phytochemical analysis (Yılmaz et al., 2020). These techniques enable the detailed characterization of complex plant extracts and the identification of novel bioactive compounds with potential therapeutic applications (Petrović et al., 2019; Yılmaz et al., 2020).

 

The integration of these technological and methodological advances into pharmacological research can lead to the discovery of new drug leads and the development of more effective and safer therapeutic agents derived from aromatic medicinal plants. These advancements also provide a better understanding of the mechanisms of action of these compounds, which is crucial for their clinical application (Petrović et al., 2019; Yılmaz et al., 2020).

 

8.3 Policy and funding considerations

Policy decisions play a crucial role in shaping the research and development landscape for aromatic medicinal plants. Supportive policies that promote the conservation and sustainable use of these plants can enhance research efforts and ensure the availability of plant resources for future studies (Ebrahimi et al., 2020; Šarčević-Todosijević et al., 2023). Additionally, policies that encourage collaboration between academic institutions, industry, and traditional medicine practitioners can facilitate the translation of research findings into practical applications (Šarčević-Todosijević et al., 2023).

 

Securing funding for research on aromatic medicinal plants remains a significant challenge. While there are funding opportunities available from government agencies, non-profit organizations, and industry, competition for these funds is often intense (Ebrahimi et al., 2020; Šarčević-Todosijević et al., 2023). Moreover, the interdisciplinary nature of this research requires substantial investment in advanced technologies and collaborative efforts, which can be financially demanding.

 

To support the advancement of research on aromatic medicinal plants, it is essential to advocate for increased funding and policy support. This includes promoting the importance of this research to policymakers and funding agencies, highlighting its potential to contribute to drug discovery and public health. Additionally, establishing dedicated funding programs and research centers focused on medicinal plant research can provide the necessary resources and infrastructure to drive innovation in this field.

 

9 Concluding Remarks

This study has explored the pharmacological effects of aromatic medicinal plants, focusing on their active ingredients and mechanisms of action. It was found that essential oils and other extracts from aromatic plants have shown significant antiproliferative potential against various cancer types. The diversity and high availability of these plants make them promising candidates for cancer chemoprevention (Fitsiou and Pappa, 2019). The pharmacological activities of medicinal plants are largely attributed to secondary metabolites, such as polyphenols, which exhibit antioxidant and anti-inflammatory effects. These properties contribute to the beneficial health effects of these plants (Šarčević-Todosijević et al., 2023). The findings underscore the need for further research to isolate and characterize the active principles responsible for the observed biological activities. Understanding the molecular mechanisms of these compounds will be crucial for developing new therapeutic agents (Fitsiou and Pappa, 2019; Ganaie, 2021).

 

The demonstrated anticancer, antibacterial, and antimutagenic properties of aromatic medicinal plants suggest their potential as cost-effective and accessible sources of therapeutic agents. These plants could be integrated into traditional and modern medical practices to enhance treatment efficacy and reduce side effects (Fitsiou and Pappa, 2019; Ganaie, 2021; Šarčević-Todosijević et al., 2023). Aromatic medicinal plants possess a wide range of pharmacological activities due to their diverse and potent active ingredients. These plants offer promising avenues for developing new treatments for cancer, bacterial infections, and other diseases. Researchers should focus on isolating and characterizing the active compounds in these plants to better understand their mechanisms of action. Healthcare practitioners should consider incorporating these natural products into treatment regimens, given their efficacy and low toxicity.

 

A comprehensive evaluation of aromatic medicinal plants highlights their significant potential in medical and therapeutic applications. With their rich history and proven pharmacological properties, aromatic medicinal plants hold great promise for the future of medicine. By combining traditional knowledge with modern scientific research, we can unlock new therapeutic potentials and contribute to more holistic and sustainable medical practices.

 

Acknowledgments

This work was completed at the Traditional Chinese Medicine Research Center of Cuixi Academy of Biotechnology. The author would like to thank Dr. X.J. Fang, the director of Cuixi Academy of Biotechnology for his guidance and systematic suggestions on the draft of the paper. Thanks are also extended to the two anonymous peer reviewers for their rigorous review and valuable questions, which made the paper more systematic and complete.

 

Funding

This project was supported by the Cuixi Academy of Biotechnology's Undergraduate Vacation Study Program (Grant No. 20230709).

 

Conflict of Interest Disclosure

The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.

 

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