Analysis of Plant-Plant Interactions Reveals the Presence of Potent Antileukemic Compounds.

A method to identify anticancer compounds in plants was proposed based on the hypothesis that these compounds are primarily present in plants to provide them with an ecological advantage over neighboring plants and other competitors. According to this view, identifying plants that contain compounds that inhibit or interfere with the development of other plant species may facilitate the discovery of novel anticancer agents. The method was developed and tested using Magnolia grandiflora, Gynoxys verrucosa, Picradeniopsis oppositifolia, and Hedyosmum racemosum, which are plant species known to possess compounds with cytotoxic activities. Plant extracts were screened for growth inhibitory activity, and then a thin-layer chromatography bioautography assay was conducted. This located the major antileukemic compounds 1, 2, 4, and 5 in the extracts. Once the active compounds were located, they were extracted and purified, and their structures were determined. The growth inhibitory activity of the purified compounds showed a significant correlation with their antileukemic activity. The proposed approach is rapid, inexpensive, and can easily be implemented in areas of the world with high biodiversity but with less access to advanced facilities and biological assays.

Distinguishing Vaccinium species by chemical fingerprinting based on NMR spectra, validated with spectra collected in different laboratories.

A method was developed to distinguish Vaccinium species based on leaf extracts using nuclear magnetic resonance spectroscopy. Reference spectra were measured on leaf extracts from several species, including lowbush blueberry (Vaccinium angustifolium), oval leaf huckleberry (Vaccinium ovalifolium), and cranberry (Vaccinium macrocarpon). Using principal component analysis, these leaf extracts were resolved in the scores plot. Analysis of variance statistical tests demonstrated that the three groups differ significantly on PC2, establishing that the three species can be distinguished by nuclear magnetic resonance. Soft independent modeling of class analogies models for each species also showed discrimination between species. To demonstrate the robustness of nuclear magnetic resonance spectroscopy for botanical identification, spectra of a sample of lowbush blueberry leaf extract were measured at five different sites, with different field strengths (600 versus 700 MHz), different probe types (cryogenic versus room temperature probes), different sample diameters (1.7 mm versus 5 mm), and different consoles (Avance I versus Avance III). Each laboratory independently demonstrated the linearity of their NMR measurements by acquiring a standard curve for chlorogenic acid (R(2) = 0.9782 to 0.9998). Spectra acquired on different spectrometers at different sites classifed into the expected group for the Vaccinium spp., confirming the utility of the method to distinguish Vaccinium species and demonstrating nuclear magnetic resonance fingerprinting for material validation of a natural health product.

High resilience of Pacific Island forests to a category- 5 cyclone.

Assessing how forests respond to, and recuperate from, cyclones is critical to understanding forest dynamics and planning for the impacts of climate change. Projected increases in the intensity and frequency of severe cyclones can threaten both forests and forest-dependent communities. The Pacific Islands are subject to frequent low-intensity cyclones, but there is little information on the effects of high intensity cyclones, or on how forest stewardship practices may affect outcomes. We assess the resistance and resilience of forests in three community-stewarded sites on the island of Tanna, Vanuatu, to the wind-related effects of 2015 Category-5 Cyclone Pam, one of the most intense cyclones to make landfall globally. Drawing on transect data established pre-and post-cyclone, we (1) test whether windspeed and tree structural traits predict survival and damage intensity, and whether this varies across sites; (2) assess post-cyclone regeneration of canopy, ground cover, seedlings, and saplings, and how community composition shifts over time and across sites. In sites that sustained a direct hit, 88 % of trees were defoliated, 34 % sustained severe damage, and immediate mortality was 13 %. Initial mortality, but not severe damage, was lower in areas that received an indirect hit and had lower windspeed. Larger trees and those with lighter wood had a higher probability of uprooting and snapping, respectively. Canopy and ground cover regenerated within three years and seedling and sapling regeneration was widespread across life histories, from pioneer to mature forest species. Three species of non-native vines recruited post-cyclone but within 5 years had largely declined or disappeared with canopy closure. Tanna's historical cyclone frequency, combined with customary stewardship practices that actively maintain a diversity of species and multiplicity of regeneration pathways, are likely responsible for the island's resistance and resilience to an intense tropical cyclone.

Costus spicatus tea failed to improve diabetic progression in C57BLKS/J db/db mice, a model of type 2 diabetes mellitus.

AIM OF THE STUDY: Costus spicatus Sw. (Costaceae) is a prominent medicinal herb used by Dominicans in the Dominican Republic and the United States for the treatment of diabetes, a growing epidemic in the Hispanic community. An ethnobotanical survey of the Dominican community in New York City revealed the popular use of a tea from the insulina plant to treat hyperglycemia. Insulina was identified as Costus spicatus. We tested the ability of a tea made from the leaves of Costus spicatus to alter glucose homeostasis in C57BLKS/J (KS) db/db mice, a model of obesity-induced hyperglycemia with progressive beta cell depletion. MATERIALS AND METHODS: From 6 to 16 weeks of age, Experimental and Control animals (n=6/5) were given ad lib access to Costus spicatus tea or water, respectively. RESULTS: Weight gain and progression of hyperglycemia and insulinopenia between the Experimental and Control groups were statistically indistinguishable. There was no difference between groups in average fed or fasting glucose and insulin concentrations. Intraperitoneal (IP) insulin tolerance testing after the 10-week study period showed that Costus spicatus tea consumption did not alter insulin sensitivity. CONCLUSIONS: These data suggest that at the dose given, tea made from Costus spicatus leaves had no efficacy in the treatment of obesity-induced hyperglycemia. More investigation is needed to more fully explore dosages and the possible utility and biological activity of this common Dominican herbal remedy for the treatment of type 2 diabetes mellitus.

Identifying Phytochemicals from Biomedical Literature Utilizing Semantic Knowledge Sources.

Chemicals derived from plants (phytochemicals) are major concepts of interest in the study of medicinal plants. To date, efforts to catalogue and organize phytochemical knowledge have resorted to manual approaches. This study explored the potential to leverage publicly accessible semantic knowledge sources for identifying possible phytochemicals. Within the context of this feasibility study, putative phytochemicals were identified for more than 4,000 plants from the Medical Subject Headings Supplementary Concept Records and the Semantic MEDLINE Database. An examination of phytochemicals identified for five selected plant species using the method developed here reveals that there is a disparity in electronically catalogued phytochemical knowledge compared to information from Dr. Duke's Phytochemical and Ethnobotanical Databases maintained by the United States Department of Agriculture. The results therefore suggest that semantic knowledge sources for biomedicine can be utilized as a source for identifying potential phytochemicals and thus contribute to the overall curation of plant phytochemical knowledge.

Traditional lifestyles, transition, and implications for healthy aging: An Example from the remote island of Pohnpei, Micronesia.

Lifestyle-related, non-communicable diseases, such as diabetes, hypertension, and obesity have become critical concerns in the Pacific islands of Micronesia. We investigated the relationship between the diminution of traditional lifestyle practices and the decline in the health of the population in the State of Pohnpei, Federated States of Micronesia. To assess this, our interdisciplinary team developed two scales, one to rank individuals on how traditional their lifestyles were and one to rank individuals on the healthiness of their lifestyles. Participants' locations were categorized as living on a remote atoll, living on the main island, or as a transitional population. Pohnpeians living in transitional communities (e.g. recently moved from a remote atoll to the main island, or the reverse) ranked lowest on both the tradition and health scales, rather than ranking intermediate between the remote and main island groups as we had hypothesized. As predicted, individuals residing on the remote atolls were living the most traditional lifestyles and also had the healthiest lifestyles, based on our rating system. The higher an individual scored on the tradition scale, e.g. the more traditional life they lived, the higher they scored on the health scale, suggesting the importance of traditional lifestyle practices for maintaining health. These findings have significant implications for promoting health and longevity of Micronesians and other Pacific Island peoples. We suggest the process of transition be recognized as a significant lifestyle and health risk and be given the attention we give to other risk factors that negatively influence our health. Based on our findings, we discuss and recommend the revitalization of particular traditional lifestyle practices, which may advance healthy aging among Pohnpeians.

Feeling the pulse in Maya medicine: an endangered traditional tool for diagnosis, therapy, and tracking patients' progress.

Throughout history, diagnostic tools utilizing the human senses, such as pulse diagnosis, have developed all over the world. In many areas where medical technology is limited or absent, they persist, whereas in other areas these skills are in danger of extinction. The practice of pulse diagnosis by the accomplished Maya healer, Don Elijio Panti, who lived in Belize, Central America, was observed over the final decade of his life and work. Don Elijio used pulse palpation as a diagnostic tool, therapeutic tool, and as a means for tracking patients' progress. He could diagnose a wide array of both physical and spiritual afflictions and was observed diagnosing 42 different conditions or states throughout this period by feeling the pulse. He recognized at least 28 distinct pulse types. Herein, the authors report the detailed system of an endangered diagnostic tradition as practiced by the late, acclaimed Maya healer, including pulse-type descriptions and corresponding diagnoses. Pulse diagnosis is still practiced today among some of Belize's diminishing population of traditional healers, although no practice appears to be as developed as that of the previous generation of Maya healers. Furthermore, it is unlikely that there are new practitioners of pulse diagnosis in the Maya community to maintain and build on the disappearing tradition. Given the unfortunate paucity of data on Maya pulse diagnosis, the practice of pulse diagnosis in Traditional Chinese Medicine (TCM) is used as an illustrative framework for documenting Don Elijio's practice. Corresponding diagnoses from TCM and Don Elijio's system are compared, elucidating similarities between the two disparate medical systems.

Harnessing Biomedical Natural Language Processing Tools to Identify Medicinal Plant Knowledge from Historical Texts.

The growing amount of data describing historical medicinal uses of plants from digitization efforts provides the opportunity to develop systematic approaches for identifying potential plant-based therapies. However, the task of cataloguing plant use information from natural language text is a challenging task for ethnobotanists. To date, there have been only limited adoption of informatics approaches used for supporting the identification of ethnobotanical information associated with medicinal uses. This study explored the feasibility of using biomedical terminologies and natural language processing approaches for extracting relevant plant-associated therapeutic use information from historical biodiversity literature collection available from the Biodiversity Heritage Library. The results from this preliminary study suggest that there is potential utility of informatics methods to identify medicinal plant knowledge from digitized resources as well as highlight opportunities for improvement.

Identifying Plant-Human Disease Associations in Biomedical Literature: A Case Study.

The impact of ethnobotanical data from surveys of traditional medicinal uses ofplants can be enhanced through the validation of biomedical knowledge that may be embedded in literature. This study aimed to explore the use of informatics approaches, including natural language processing and terminology resources, for extracting and comparing ethnobotanical leads from biomedical literature indexed in MEDLINE. Using ethnobotanical data for plant species described in Primary Health Care Manuals of the Micronesian islands of Palau and Pohnpei, the results of this study were done relative to disease concepts from the "Mental, Behavioral And Neurodevelopmental Disorders " ICD-9-CM category. The results from this feasibility study suggest that informatics methods can be used to extract and prioritize relevant ethnobotanical information from biomedical knowledge literature.

A traditional method of Cinnamomum carolinense preparation eliminates safrole from a therapeutic Pohnpean tea.

Cinnamomum carolinense, locally known as madeu, is a tree endemic to the volcanic mountains of the Island of Pohnpei in the Eastern Carolines of the South Pacific. The bark is harvested from trees and brewed to make a medicinal tea and hot beverage that is regularly consumed. Many species of Cinnamomum contain the known hepatocarcinogen safrole, sparking concern regarding habitual consumption of this beverage. HPLC-PDA analysis confirmed the presence of the carcinogen in alcoholic extracts of Cinnamomum carolinense bark shavings (0.435%, w/w), but safrole was not detected in the tea. The limit of detection and limit of quantitation of safrole were determined to be 1.25 and 3.75 microg/mL, respectively. The traditional preparation method, which boils the bark shavings, degrades the safrole.

Traditional preparation of Phaleria nisidai, a Palauan tea, reduces exposure to toxic daphnane-type diterpene esters while maintaining immunomodulatory activity.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Phaleria nisidai Kaneh. (Thymelaeaceae) are brewed into a tea commonly used as a tonic, strengthening beverage and immune enhancer in Palau, Micronesia. Recently, the leaves of P. nisidai have been shown to contain toxic daphnane diterpene esters which may pose a public health threat to Palauans. AIMS OF THE STUDY: This project documents the use frequency, preparation and side effects of P. nisidai. The content of daphnane diterpene esters in aqueous and methanol extracts and infusions prepared by healers in Palau is compared to assess the risk of daphnane ingestion associated with traditional consumption. Quantitative results are correlated with an in vitro assessment of the immunomodulating activity of the extracts. MATERIALS AND METHODS: Research participants, comprising traditional healers and laypeople, were interviewed concerning use patterns and side effects of P. nisidai. Several traditional healers prepared and provided boiled tea samples for chemical analysis. Leaves were collected and methanolic and aqueous extractions were prepared in the laboratory. Peripheral blood mononuclear cells (PBMCs) were cultured with various concentrations of methanol and aqueous leaf extracts and their output of IFNγ was measured using ELISA. Cell proliferation was also assessed using the MTT assay. The concentration of selected daphnane diterpene esters in healer-prepared infusions, lab methanol and lab aqueous extracts was quantified using ultraperformance liquid chromatography-mass spectrometry-triple quadrupole detection (UPLC-MS-TQD). RESULTS: Through structured interviews it was determined that P. nisidai tea was used frequently, with many participants drinking it daily. The reported side effects were mild, and with the exception of diarrhea (n=2), no side effect was mentioned more than once. Methanol extracts contained 4.0µg simplexin, 17.6µg acetoxyhuratoxin and 2.3µg huratoxin per g dry leaf material. In traditional water infusions provided by healers and in standardized lab-prepared aqueous extracts all three compounds were below the limit of detection (16.3ng/mL) using our UPLC-MS-TQD method. Methanol and aqueous extracts increased the release of IFNγ by PBMCs (p<0.05); however, methanol extracts were significantly more active than aqueous extracts (p<0.05). Methanol and aqueous extracts significantly increased proliferation of PBMCs, causing at least 60% more cell proliferation than negative control (p<0.05). CONCLUSIONS: The presence of daphnane diterpene esters in a frequently consumed traditional beverage was initially viewed as a public health concern, though interview data reveal that Palauans do not observe toxicity or side effects associated with their use of P. nisidai tea. Concurrently, daphnanes are present in methanolic extracts but not detected in aqueous preparations indicating that the traditional method of preparation avoids the extraction of these potentially toxic compounds, while still maintaining immunostimulant activity.

An ethnobotany of the Lukomir Highlanders of Bosnia & Herzegovina.

BACKGROUND: This aim of this study is to report upon traditional knowledge and use of wild medicinal plants by the Highlanders of Lukomir, Bjelasnica, Bosnia and Herzegovina (B&H). The Highlanders are an indigenous community of approximately 60 transhumant pastoralist families who speak Bosnian (Bosanski) and inhabit a highly biodiverse region of Europe. This paper adds to the growing record of traditional use of wild plants within isolated communities in the Balkans. METHODS: An ethnobotanical study using consensus methodology was conducted in Lukomir in Bjelasnica's mountains and canyons. Field work involved individual semi-structured interviews during which informants described plants, natural product remedies, and preparation methods on field trips, garden tours, while shepherding, or in settings of their choice. Plant use categories were ranked with informant consensus factor and incorporated into a phylogenetic tree. Plants cited were compared to other ethnobotanical surveys of the country. RESULTS: Twenty five people were interviewed, resulting in identification of 58 species (including two subspecies) from 35 families, which were cited in 307 medicinal, 40 food, and seven material use reports. Individual plant uses had an average consensus of five and a maximum consensus of 15 out of 25. There were a number of rare and endangered species used as poisons or medicine that are endemic to Flora Europaea and found in Lukomir. Ten species (including subspecies) cited in our research have not previously been reported in the systematic ethnobotanical surveys of medicinal plant use in B&H: (Elymus repens (L.) Gould, Euphorbia myrsinites L., Jovibarba hirta (L.) Opiz, Lilium bosniacum (Beck) Fritsch, Matricaria matricarioides (Less.) Porter ex Britton, Phyllitis scolopendrium (L.) Newman, Rubus saxatilis L., Silene uniflora Roth ssp. glareosa (Jord.) Chater & Walters, Silene uniflora Roth ssp. prostrata (Gaudin) Chater & Walters, Smyrnium perfoliatum L.). New uses not reported in any of the aforementioned systematic surveys were cited for a total of 28 species. Thirteen percent of medicinal plants cited are endemic: Helleborus odorus Waldst. et Kit., Gentiana lutea L., Lilium bosniacum (Beck) Fritsch, Silene uniflora Roth ssp. glareosa (Jord.) Chater & Walters., Silene uniflora Roth ssp. prostrata (Gaudin) Chater & Walters, Salvia officinalis L., Jovibarba hirta (L.) Opiz, and Satureja montana L. CONCLUSIONS: These results report on the cohesive tradition of medicinal plant use among healers in Lukomir, Bosnia and Herzegovina. This work facilitates the community's development by facilitating local and international conversations about their traditional medicine and sharing insight for conservation in one of Europe's most diverse endemic floristic regions, stewarded by one of Europe's last traditional Highland peoples.

Biochemical and evolutionary aspects of arthropod predation on ferns.

The widely held assumption that very few arthropods feed on ferns was questioned following field observations of arthropod damage on ferns in the state of Veracruz, Mexico. The extent and type of damage was recorded and it was found that in a measured locality, ferns were no less attacked than the angiospermous flora. As chemistry and arthropod host relationships have been shown to be so closely intertwined, plants collected in the field were analysed for both condensed tannins and cyanogenic glycosides, compounds known to be effective deterrents in temperate climates. Although all ferns tested contained tannins these did not appear to inhibit predation. Cyanogenic glycosides were present in only 3% of the fern species analysed, and it is, therefore, unlikely that they play a significant role as defensive compounds in the ferns examined.A literature search revealed a large number of ferns cited as being arthropod hosts. Approximately 420 named species of arthropods have been recorded, the majority of which are from the orders Coleoptera, Hymenoptera, Lepidoptera, and Hemiptera. Both evolutionary primitive (sawflies) and advanced (moths) arthropods are reported to be present on ferns suggesting possible coevolution of arthropods and ferns both before and after the radiation of angiosperms.

Alseis yucatanensis: a natural product from Belize that exhibits multiple mechanisms of vasorelaxation.

An aqueous extract of the bark of Alseis yucatanensis was studied to determine its mechanism of action in the relaxation of endothelium-denuded rat aortic tissues. The extract relaxed both norepinephrine (NE) and KCl-contracted vessels, with ED(50)'s of 0.12 and 1.73 mg/mL, respectively. In NE-contracted vessels, two phases of relaxation were evident which were separated in both time and dose range. At high concentrations, a rapid relaxation was seen that was due to the blocking of internal (ED(50)=0.49 mg/mL) and external (ED(50)=2.34 mg/mL) calcium channels. A second, slowly developing (i.e., long-term) relaxation to baseline was seen at lower concentrations. The time to complete relaxation was dose-dependent. This long-term response was not seen in KCl-contracted vessels, was prolonged by TEA, and could be reversed by the addition of KCl to the bath. These data suggest that the long-term relaxation is due to the opening of potassium channels.

Double incision wound healing bioassay using Hamelia patens from El Salvador.

Hamelia patens Jacq. (Rubiaceae) has received little attention in the laboratory for its wound healing ability even though it is commonly used as a treatment for wounds throughout Central America. A double incision wound healing bioassay was carried out with a crude extract of Hamelia patens collected from El Salvador. Animals were divided into three groups. Group I (n = 14) had the left incision treated with 5% (w/w) Hamelia patens and the contralateral side with petroleum jelly (PJ). Group II (n = 14) had the left incision treated with 10% (w/w) ointment and the contralateral side with petroleum jelly. Group III (n = 10) had the left incision treated with petroleum jelly and the contralateral side left untreated. Breaking strength of the incisions was measured on day 7 and day 12. For Groups I and II, there was no significant difference between treatment and control incisions at day 7. On day 12, there was a significant difference between the treated and control incisions for Groups I and II. There was no significant difference between petroleum jelly and untreated incisions for Group III on day 7 and day 12. Hamelia patens does increase breaking strength of wounds significantly more than the control group. Further wound healing studies of this plant are warranted.

Ethnobotanical literature survey of medicinal plants in the Dominican Republic used for women's health conditions.

This ethnobotanical literature survey is part of an on-going study in New York City investigating Dominican and Chinese healing systems and the herbal treatments used for the following women's conditions: uterine fibroids (benign tumors of uterine smooth muscle); menorrhagia (excessive uterine bleeding); endometriosis (growth of endometrial tissue outside of the uterus); and hot flashes (sudden brief sensations of heat commonly experienced during menopause). The objectives of this survey were: (1) to search literature on medicinal plants used in the Dominican Republic and identify those used for the above listed conditions and their symptoms; (2) to compare the use between herbal treatments reported in the literature with those prescribed by Dominican healers in New York City; and (3) to evaluate the extent to which healers may have changed their use of plants in order to adapt to availability in the New York City environment. A total of 87 plant species were reported in the Dominican literature for these conditions and symptoms. Nineteen species overlapped from the literature survey and the fieldwork with Dominican healers in New York City, representing 29% (n=65) of the plants prescribed by healers in New York City. This study offers a model to investigate changes in plant use as people migrate to urban centers where they are surrounded by diverse cultures, healing systems, and new environments.

Does the name really matter? The importance of botanical nomenclature and plant taxonomy in biomedical research.

ETHNOPHARMACOLOGICAL RELEVANCE: Medical research on plant-derived compounds requires a breadth of expertise from field to laboratory and clinical skills. Too often basic botanical skills are evidently lacking, especially with respect to plant taxonomy and botanical nomenclature. Binomial and familial names, synonyms and author citations are often misconstrued. The correct botanical name, linked to a vouchered specimen, is the sine qua non of phytomedical research. Without the unique identifier of a proper binomial, research cannot accurately be linked to the existing literature. Perhaps more significant, is the ambiguity of species determinations that ensues of from poor taxonomic practices. This uncertainty, not surprisingly, obstructs reproducibility of results-the cornerstone of science. MATERIALS AND METHODS: Based on our combined six decades of experience with medicinal plants, we discuss the problems of inaccurate taxonomy and botanical nomenclature in biomedical research. This problems appear all too frequently in manuscripts and grant applications that we review and they extend to the published literature. We also review the literature on the importance of taxonomy in other disciplines that relate to medicinal plant research. RESULTS AND DISCUSSION: In most cases, questions regarding orthography, synonymy, author citations, and current family designations of most plant binomials can be resolved using widely-available online databases and other electronic resources. Some complex problems require consultation with a professional plant taxonomist, which also is important for accurate identification of voucher specimens. Researchers should provide the currently accepted binomial and complete author citation, provide relevant synonyms, and employ the Angiosperm Phylogeny Group III family name. Taxonomy is a vital adjunct not only to plant-medicine research but to virtually every field of science. CONCLUSIONS: Medicinal plant researchers can increase the precision and utility of their investigations by following sound practices with respect to botanical nomenclature. Correct spellings, accepted binomials, author citations, synonyms, and current family designations can readily be found on reliable online databases. When questions arise, researcher should consult plant taxonomists.

Merging bioactivity with liquid chromatography-mass spectrometry-based chemometrics to identify minor immunomodulatory compounds from a Micronesian adaptogen, Phaleria nisidai.

This study presents a strategy based on repeatable reversed-phase LC-TOF-MS methods and chemometric statistical tools, including untargeted PCA and supervised OPLS-DA models, to identify low-yielding compounds with potent immunostimulant activity in Phaleria nisidai (Thymelaeaceae), a plant with a history of use as an adaptogen on the islands of Palau in Micronesia. IFNγ ELISA assays were used to classify chromatographic fractions according to imunomodulatory activity prior to LC-TOF-MS chemometric analysis to target and identify compounds likely to contribute to observed activity. Simplexin, a daphnane diterpene ester, was identified for the first time from this genus and caused an increase in the production of cytokines (IFNγ, IL1ß, IL6, and IL13) by peripheral blood mononuclear cells. Five other daphnane diterpene esters were tentatively identified for the first time from this plant based on mass spectral data and are marker metabolites distinguishing active from inactive fractions. This analytical approach increased the efficiency of bioactivity-guided fractionation and has the potential to minimize redundant isolation and identify minor constituents with potent activity from a complex matrix.

Stability of black cohosh triterpene glycosides and polyphenols: potential clinical relevance.

Concurrent to a clinical trial of black cohosh for menopausal hot flashes, the long-term stability of the black cohosh, over the duration of the clinical trial, was examined. Analytical results showed that the major constituents, both triterpene glycosides and polyphenols, were stable over the 3-year period of testing. These results indicate that a black cohosh product stored for several years in a controlled environment does not undergo significant changes in its major constituents. These results have implications not only for clinical research in natural products, but for basic science, as well as the dietary supplements industry.