Search

The Asteraceae is the largest family of plants in North America and is widely used as medicine by Indigenous peoples. This study investigated the medicinal ethnobotany of North American Asteraceae to identify taxa that appear preferentially selected or avoided for general and specific medicinal uses. Asteraceae-specific ethnobotanical reports recorded in the Native American Ethnobotany database were compiled and, using residual and binomial analyses, 14 tribes were compared and ranked as either over- or under-selected for medicine, food, or technology, and for different categories of medicinal applications. Statistical analysis supported the hypothesis that the selection of species for ethnobotanical purposes is non-random and does not depend on the size of the flora. The Anthemideae tribe was identified as over-selected for all types of applications, including most therapeutic categories, most significantly as pulmonary and orthopedic aids. Subsequent analysis revealed that the over-representation of this tribe was attributed mainly to Achillea millefolium L. and Artemisia spp. The significance of Anthemideae, particularly of Achillea and Artemisia species as highly selected medicinal taxa, emphasizes their cultural importance to Indigenous North Americans. Residual and binomial statistics generally provided parallel results, but supplementary statistical methods, further in-depth investigation of other use categories, and inclusion of plant distribution data may provide greater insight into traditional uses of Asteraceae in North America.

Cl-AlPc-based OTFTs can be used as low-cost, point-of-source cannabinoid sensors. Through device performance and film characterization, the film thickness and device geometry has been optimized for both solution and vapor THC samples.

The 17th Annual Natural Health Products Research Conference hosted by the NHP Research Society of Canada (NHPRS) will be held from June 7–9 & 14–16, 2021, virtually hosted by the University of Ottawa, in Ottawa, Ontario. Founded in 2003 by a collaboration of academic, industry, and government researchers from across Canada, the NHPRS is a Canadian federally incorporated non-profit organization. The goals of the NHPRS are; (a) to promote scientifically rigorous research and education on natural health products (NHPs), (b) to develop a national research community that encompasses academic, health professional, government and a broad-base of industry stakeholders, (c) to support national research priorities that best enable the informed and appropriate use of NHPs that are safe and efficacious, (d) to increase the capacity for NHP research and education, (e) to facilitate effective NHP knowledge transfer and translation, (f) to support the use of science-based product quality standards and the use of well-characterized materials and protocols in research and (g) to foster value-chain development through interdisciplinary NHP research collaborations and networking. To forward these objectives, the NHPRS has held annual research conferences since inception with themes that highlight the important trends in the ever growing and changing field of NHP Research. The theme of the 17th Annual Conference is “Canada and the Changing Global NHP Landscape”, featuring six "live" sessions (Global Stewardship and Sustainability, NHPs in the Time of the Pandemic, Indigenous Traditional Medicine, Global Advances in NHP Research, NHPs in Clinical Practice and Research, and Regulation, Policy, Commerce) and contributed sessions on topics such as cancer, cannabis, functional foods. natural product chemistry, and more. The abstracts are ordered alphabetically by presenting author last name, divided by topic sessions. For more information, please visit: http://www.nhprs.ca/.

Introduction: Methylphenidate is a psychostimulant prescribed for symptoms of attention-deficit hyperactivity disorder (ADHD), and is metabolized by carboxylesterase-1 (CES1) enzyme. There is little information on herbal medicine used for ADHD and their interactions with CES1. Methods: An in vitro study was conducted with extracts of 21 herbal medicines using a recombinant human CES1 enzyme inhibition assay to determine if the extracts influence enzyme activity. Extracts were initially evaluated for their potential to inhibit CES1 metabolism of 4-nitrophenyl acetate using an in vitro Microtiter plate enzyme inhibition assay. The most active extracts were serially diluted to establish the half maximal inhibitory concentration (IC50) and were then tested for potential irreversible inhibition using a time-dependence enzyme assay. Phytochemical characterization of selected extracts was performed, and marker compounds were evaluated for their potential to inhibit CES1-mediated metabolism of the probe substrate. Results: The extracts exhibited a range of inhibition of CES1 activity, ranging from 15–95% at a standard screening concentration of 200 ?g/mL. Rhodiola rosea was the most potent inhibitor of CES1 (IC50 = 4.7 ?g/mL). No time-dependent inhibitors of CES1 were identified in this study. At 10 ?g/mL, marker compounds of ginger, [8]-gingerol (60.3%) and [10]-gingerol (67.2%), showed significant (P < 0.05) inhibition of CES1 compared to vehicle control. Discussion: The herbal medicine extracts showed varying inhibition at supraphysiologic concentrations and no risk of mechanism-based inhibition was observed. Herbal medicine used by ADHD patients have the potential to interact with CES1-mediated metabolism. To mitigate potential risk of herb-drug interactions, patients using herbal medicine alongside CES1-metabolized drugs should discuss their use with healthcare professionals. Conclusions: In vivo and clinical research are required to determine if the potential interactions would significantly affect safety and efficacy of methylphenidate at a clinical level.

Recent research demonstrates that Echinacea possesses cannabimimetic activity with potential applications beyond common contemporary uses for relief of cold and flu symptoms. In this study, we investigated the in vitro inhibitory effect of root extracts of Echinacea purpurea and Echinacea angustifolia on fatty acid amide hydrolase, the main enzyme that degrades the endocannabinoid anandamide. The objective was to relate variation in bioactivity between commercial Echinacea genotypes to their phytochemical profiles and to identify determinants of activity using biochemometric analysis. Forty root extracts of each of species were tested for inhibition of fatty acid amide hydrolase and analyzed by HPLC-DAD/MS to identify and quantitate alkylamides and caffeic acid derivatives. Fatty acid amide hydrolase inhibition ranged from 34 – 80% among E. angustifolia genotypes and from 33 – 87% among E. purpurea genotypes. Simple linear regression revealed the caffeic acid derivatives caftaric acid and cichoric acid, and the alkylamide dodeca-2E,4Z-diene-8,10-diynioc acid 2-methylbutylamide, as the strongest determinants of inhibition in E. purpurea (r* = 0.53, 0.45, and 0.20, respectively) while in E. angustifolia, only CADs were significantly associated with activity, most notably echinacoside (r* = 0.26). Regression analysis using compound groups generated by hierarchical clustering similarly indicated that caffeic acid derivatives contributed more than alkylamides to in vitro activity. Testing pure compounds identified as determinants of activity revealed cichoric acid (IC50 = 45 ± 4 µM) and dodeca-2E,4E,8Z,10E-tetraenoic acid isobutylamide (IC50 = 54 ± 2 µM) as the most active. The results suggest that several phytochemicals may contribute to Echinaceaʼs cannabimimetic activity and that ample variation in genotypes exists for selection of high-activity germplasm in breeding programs.

Modern Natural Health Products (NHPs) are medicinal dosage forms also known and regulated variously as Dietary Supplements, Phytomedicines and Complementary and Alternative Medicines in different countries. Related regulated food products include Medicinal Foods, Supplemented Foods, Functional Foods, and Nutraceuticals, while cannabis products are regulated separately. All of the above are derived from or inspired by natural products that have existed for millennia. Most have been developed through ancient medical systems (e.g. Traditional Chinese Medicine and Ayurveda) and distinct indigenous cultures, which combine worldview, beliefs, practices, into “ways of knowing”. Others were identified or advanced through scientific discovery and biotechnology. Together, they have contributed—and will continue to contribute—to human health and economy at a global scale.

Organic thin-film transistors (OTFTs) have shown promise for a range of sensing applications, with phthalocyanine-based OTFTs demonstrated as sensors for atmospheric parameters, volatile gases, and small organic molecules including cannabinoids. However, the process of fabricating, testing, and optimizing OTFTs in a laboratory setting requires highly specialized equipment, materials, and expertise. To determine if sensor development can be expedited and thus reduce manufacturing burden, spectroelectrochemistry is applied to rapidly screen for molecular interactions between metal-free phthalocyanines and a variety of metal phthalocyanines (MPcs) and the cannabinoids Δ9-tetrahydrocannabinol (THC) or cannabidiol (CBD), with and without a cannabinoid-sensitive chromophore (Fast Blue BB). Spectral analyses are corroborated by 2D-NMR and related to measured OTFT performance. Spectroelectrochemical changes to the Q band region of the phthalocyanine spectra in the presence of analytes can be used to predict the response of OTFTs. Thus, with spectroelectrochemistry, a range of potential materials for OTFT small organic molecule-sensing applications can be quickly analyzed, and phthalocyanines with a preferred response can be selected.

Quality control is imperative for Cannabis since the primary cannabinoids, Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD), elicit very different pharmacological effects. THC/CBD ratios are currently determined by techniques not readily accessible by consumers or dispensaries and which are impractical for use in the field by law-enforcement agencies. CuPc- and F16-CuPc-based organic thin-film transistors have been combined with a cannabinoid-sensitive chromophore for the detection and differentiation of THC and CBD. The combined use of these well-characterized and inexpensive p- and n-type materials afforded the determination of the CBD/THC ratio from rapid plant extracts, with results indistinguishable from high-pressure liquid chromatography. Analysis of the prepyrolyzed sample accurately predicted postpyrolysis THC/CBD, which ultimately influences the psychotropic and medicinal effects of the specific plant. The devices were also capable of vapor-phase sensing, producing a unique electrical output for THC and CBD relative to other potentially interfering vaporized organic products. The analysis of complex medicinal plant extracts and vapors, normally reserved for advanced analytical infrastructure, can be achieved with ease, at low cost, and on the spot, using organic thin-film transistors.

Metabolomics, the application of analytical chemistry methodologies to survey the chemical composition of a biological system, is used to globally profile and compare metabolites in one or more groups of samples. Given that metabolites are the terminal end-products of cellular metabolic processes, or 'phenotype' of a cell, tissue, or organism, metabolomics is valuable to the study of the maternal-fetal interface as it has the potential to reveal nuanced complexities of a biological system as well as differences over time or between individuals. The placenta acts as the primary site of maternal-fetal exchange, the success of which is paramount to growth and development of offspring during pregnancy and beyond. Although the study of metabolomics has proven moderately useful for the screening, diagnosis, and understanding of the pathophysiology of pregnancy complications, the placental metabolome in the context of a healthy pregnancy remains poorly characterized and understood. Herein, we discuss the technical aspects of metabolomics and review the current literature describing the placental metabolome in human and animal models, in the context of health and disease. Finally, we highlight areas for future opportunities in the emerging field of placental metabolomics.

Historical ethnobotanies of indigenous peoples of the North American prairies reveal treatment of many painful conditions by Echinacea spp. Recent evidence suggests a pharmacological basis for such use as the bioactivity of E. angustifolia and E. purpurea is mediated, in part, through activation of the endocannabinoid system (ECS). Whereas the cannabimimetic effects of individual echinacea products and alkylamides have been described, the activity of crude extracts has not been compared between cannabinoid (CB) receptors or across species or genotypes. Moreover, few studies have explored echinacea’s engagement of the ECS for historic treatments or new therapeutic applications in peripheral inflammatory pain. We hypothesized that 1) the in vitro effects of root extracts on CB receptor internalization would vary with species and phytochemistry, and that echinacea root extracts would reduce inflammatory pain in vivo through activation of the ECS. Root extracts of different E. angustifolia and E. purpurea accessions were prepared, analyzed by HPLC-DAD to quantify caffeic acid derivatives and alkylamides (AKA), and tested for agonist and antagonist activities using receptor redistribution assays. Linear regression of activity relative to phytochemistry identified predictive compounds that were assessed individually in redistribution assays. Extracts were evaluated in the Hargreaves model of chronic inflammatory pain in rats with co-administration of selective CB1/2 antagonists to gauge involvement of the ECS. CB receptor agonist activity varied among accessions of both species with linear regression revealing a significant relationship between CB1 activity and AKA2 for E angustifolia, and AKA 9 + 10 for E purpurea. CB2 activity was positively related with AKA 9 + 10 and total AKAs in E. angustifolia. Four isolated AKA demonstrated agonist activity in the CB2, but not CB1, assay. In the inflammatory pain model, oral administration of either E angustifolia or E. purpurea root extract produced dose-dependent analgesic effects that were partially reversed by co-administration of CB receptor antagonists. This study demonstrates that in vitro effects of crude echinacea root extracts on CB receptors is predicted by phytochemistry. In vivo, echinacea has potential applications for peripheral inflammatory pain such as arthritis and burns, reflecting the traditional uses of Indigenous North Americans.

The legalization of cannabis has magnified the importance of determining the quantity and identification of cannabinoids. Both industry and consumers are highly interested in the content of cannabinoids available in their products, while health care professionals and regulators are concerned with the safety of cannabis. Quantification of major cannabinoids in products answers some of these concerns. Currently, popular methods of quantifying cannabinoids use high-pressure liquid chromatography (HPLC). Still, these HPLC methods are limited to quantifying a small number of cannabinoids unless more powerful but more costly instruments are employed to achieve better analysis, such as UHPLC and mass spectrometry. We propose a quick method that successfully separates and quantifies 14 cannabinoids in a single run using capillary electrophoresis (CE) coupled with a UV detector in 25 minutes. Our CE method demonstrated the limit of detection between 1.2–1.8 µg/mL, with the detection range reaching up to 50 µg/mL.

In the past decade, the predominant prohibition model for cannabis use has shifted towards a regulated legal model, most widely in the context of medical purposes. In 2018, Canada became the first G7 country to legalize cannabis for adult use, implementing a two-phase roll-out of cannabis regulations. A stated goal of the new legal framework is to minimize harms by providing a safe supply of cannabis to Canadian consumers. One way that this can be achieved is through appropriate Quality Control and Quality Assurance (QC/QA) measures. Canada has implemented stringent QC/QA measures for all classes of cannabis, which include requirements such as labelling THC and CBD content per product and limiting THC doses. This paper will provide an overview of the current QC/QA measures in Canada, highlighting differences based on class of cannabis and consider the strengths and weaknesses of the current standards. QC/QA standards represent a key safety feature that can enable informed purchasing and provide consumers with necessary information about various cannabis products. As Canada continues to progress its cannabis policies, QC/QA measures provide a key consideration for ensuring Canada meets its objective of providing a safe supply of cannabis to Canadian consumers.

Extraction of plants for drug discovery, ethnopharmacology or application as nutraceuticals and natural health products (NHPs; also known as dietary supplements, phytomedicines and complementary and alternative medicines) is described for scientists new to the field. Primary identification and handling of plant material for extraction is a key first step. Simple solvent extractions are described with routine procedures in the lab to produce crude extracts for bioassay screening or industrial use. Specialized methods targeting specific groups of compounds but rarely producing pure compounds can also be used. These methods include soxhlet, accelerated solvent, subcritical water, supercritical CO2, ultrasound, microwave, countercurrent, steam distillation and other technologies. Ultimately, researchers must identify the active principles and/or unique marker compounds in their extracts and this is accomplished by bioassay guided isolation or newer metabolomics methods. Once bioactive compounds are identified, standardized products can be produced that are reproducible and can be compared to clinically tested materials.

Context: Plants of the genus Echinacea (Asteraceae) are among the most popular herbal supplements on the market today. Recent studies indicate there are potential new applications and emerging markets for this natural health product (NHP). Objective: This review aims to synthesize recent developments in Echinacea biotechnology and to identify promising applications for these advances in the industry. Methods: A comprehensive survey of peer-reviewed publications was carried out, focusing on Echinacea biotechnology and impacts on phytochemistry. This article primarily covers research findings since 2007 and builds on earlier reviews on the biotechnology of Echinacea. Results: Bioreactors, genetic engineering and controlled biotic or abiotic elicitation have the potential to significantly improve the yield, consistency and overall quality of Echinacea products. Using these technologies, a variety of new applications for Echinacea can be realized, such as the use of seed oil and antimicrobial and immune boosting feed additives for livestock. Conclusions: New applications can take advantage of the well-established popularity of Echinacea as a NHP. Echinacea presents a myriad of potential health benefits, including anti-inflammatory, anxiolytic and antibiotic activities that have yet to be fully translated into new applications. The distinct chemistry and bioactivity of different Echinacea species and organs, moreover, can lead to interesting and diverse commercial opportunities.

Plants of the genus Echinacea have received extensive research attention, yet relatively little is known about the fruit (cypselae) of this economically important genus. The goal of this study was to investigate and compare the cypselae of four species of Echinacea: E. purpurea (L.) Moench, E. angustifolia DC., E. pallida (Nutt.) Nutt., and E. paradoxa (Norton) Britton, with respect to chemistry and germination. The phytochemical content of various fruit fractions was analyzed by high performance liquid chromatography with diode-array detection, identifying several alkylamides and caffeic-acid derivatives of medicinal value in all of the tissues. Alkylamides were most concentrated in the perianth and glands, whereas phenolics were generally enriched within the seed. Germination studies of intact cypselae and de-coated seeds demonstrate that the influence of cypsela coatings on germination depends on the growth environment. Removal of the perianth and pericarp resulted in decreased germination in soil, but significantly improved the germination of seeds of all species when sown in a sterile agar medium. In contrast, a higher percentage of intact cypselae of E. purpurea, E. angustifolia, and E. pallida germinated in soil than in sterile agar. These results suggest an ecophysiological role for phytochemicals in seed germination and will inform the cultivation and breeding of less common Echinacea species as well as the development of new products from currently unused plant material.

Ethnopharmacological relevance The Maya have traditionally used copal, Protium copal, as incense during ceremonies since pre-Columbian times. Anecdotally, copal (when burned as incense), is thought to elicit mentally uplifting and calming effects. The main objective of this study was to determine whether the incense elicits anxiolytic-like behavior in animal models using rats. A second objective was to characterize active constituents and discern potential mechanism(s) of action, specifically the involvement of the GABAergic and endocannabinoid (eCB) systems. Despite the extensive Central American use of this resin, there are currently no known scientific behavioral or pharmacological studies done with the incense. Materials and methods Quantification of the triterpenes in the copal resin and cold trapped incense was achieved by HPLC MS. Behavioral effects in rats were assessed using the elevated plus maze (EPM), social interaction (SI) test, conditioned emotion response (CER) and Novel object recognition (NOR) paradigms. Rats were exposed to burning copal (200 mg) over 5 min in a smoking chamber apparatus and then immediately tested in each behavioral paradigm. Follow-up SI tests were done using two antagonists flumazenil (1 mg/kg) and AM251 (1 mg/kg) administered systemically. Inhibition of MAGL (monoacylglycerol lipase) was measured by microplate assay with recombinant human enzyme and probe substrate. Results Phytochemical analysis revealed that copal resin and incense had high α- and β-amyrins and low lupeol triterpene content. Exposure to Protium copal incense significantly reduced anxiety-like behavior in the SI and CER tests. In contrast, no anxiolytic effects were observed in the EPM. The CER effect was time dependent. Both flumazenil and AM251 blocked the anxiolytic activity of copal revealing the involvement of GABAergic and endocannabinoid systems. Copal, as well as the identified triterpenes, potently inhibited monoacylglycerol lipase (MAGL) activity in vitro (IC50 ≤ 811 ng/mL). Conclusions This is the first study to show that copal incense from Protium copal elicits anxiolytic-like effects in fear and social interaction models as evidenced by a reduced learned fear behavior and an increase in active social interaction. It's high α and β-amyrin content suggests behavioral effects may be mediated, in part, by the known action of these terpenes at the benzodiazepine receptor. Furthermore, P. copal's observed activity through the eCB system via MAGL offers a new potential mechanism underlying the anxiolytic activity.

Background: Some pediatric patients with attention-deficit/hyperactivity disorder (ADHD) use natural health products (NHPs) such as herbal remedies. Although herbal remedies are generally considered to be safe when they are used appropriately, they may contain active components that can interact with medications being used concurrently, with potential for NHP-drug interactions leading to adverse events. Objectives: The objectives of this study were (1) to identify adverse event reports (AERs) involving commonly used herbal remedies and ADHD prescription medicines in children and adolescents; (2) to evaluate the quality of collected AERs; and (3) to assess whether NHP-drug interactions can be causally linked to reported adverse events. Methods: We systematically searched the FDAble database (FDAble.com) for herbal remedies commonly used by patients (4-18 years old) also taking ADHD drugs from 1997 to 2015. We assessed the completeness of the AERs and used three causality assessment tools modified for NHPs (Naranjo Adverse Drug Reaction Probability Scale, HORN Drug Interaction Probability Scale, and World Health Organization Uppsala Monitoring Centre Scale). Results: Of the 23 identified AERs involving both an herbal remedy and an ADHD prescription medication, most involved multiple (>3) substances with inadequate detail to assess multiple potential interactions. Following data extraction and evaluation of completeness, five AERs involving only one herbal remedy and one ADHD medication were evaluated for causality. An NHP-drug interaction was assessed to be probable in one case and to be possible in another. Both these reports involved a methylphenidate formulation and St. John's wort. Conclusions: Eighteen of the 23 identified AERs involving both an herbal remedy and an ADHD drug also involved other multiple ingredient products. The reporting quality was poor for the five AERs examined. Further research is needed to study the interaction between St. John's wort and methylphenidate.

Cannabis sativa and related products are widely used, but their potential to cause significant clinical interactions remains unclear, particularly for cannabinoid-enriched or otherwise concentrated products. The pharmacokinetics of most cannabis products is not known. Where information is known, there is wide variation. Extrapolation of limited clinical data is complicated by the complexity and variability of cannabis products as well as their delivery through various routes of administration. In vitro evidence shows that the major cannabinoids are substrates for numerous metabolic enzymes, including the cytochrome P450 metabolizing enzymes. Whereas many consumers consider cannabis products to be safe relative to alternative prescription or narcotic drugs, clinical reports of cannabis-related drug interactions and adverse events are increasing in frequency. Patients using these products, whether for medical or nonmedical purposes, together with conventional therapeutic agents may be at increased risk of adverse events, including therapeutic failure, and require enhanced monitoring.

Quantification of major cannabinoids in cannabis products is normally performed using high-pressure liquid chromatography (HPLC)-based methods. We propose a cost-effective alternative method that successfully separates and quantifies 14 cannabinoids in a single run using capillary electrophoresis (CE) coupled with a UV detector in 18 min. The separation is carried out in 60% acetonitrile in the presence of 6.5 mM sodium hydroxide and 25 µM β-cyclodextrin, resulting in good separation of cannabinoids. Our CE method demonstrated the limit of detection between 1.2–1.8 µg/mL, with the linear range reaching up to 50 µg/mL. We validated the method performance by testing a plant extract and quantifying cannabinoid content. This method is the first to separate 14 cannabinoids in one run using a CE system with UV detection.

Ethnopharmacological relevance Larix laricina , a native tree of North America, is a highly respected medicinal plant used for generations by Indigenous Peoples across its range, including the Cree of northern Quebec who use the bark to treat symptoms of diabetes. This study investigates the antioxidant capacity and bioavailability of active constituents identified in L. laricina bark extracts. Materials and methods (1) Oxygen radical absorbance capacity (ORAC) assay was employed to test antioxidant capacity of organic extracts (80% ethanol) from bark of L. laricina as well as fractions, isolated compounds, and media samples collected during permeability assays. (2) Caco-2 cell monolayer cultures were used to determine the permeability of identified antioxidants, which were quantified in basolateral media samples using liquid chromatography – tandem mass spectrometry (HPLC-ESI-MS/MS). Results Crude ethanolic extract possessed strong antioxidant potential in vitro (7.1±0.3 Trolox equivalents (TE) μM/mg). Among the 16 L. laricina fractions obtained by chromatographic separation, fraction 10 (F10) showed the highest antioxidant capacity (21.8±1.7 µm TE/mg). Among other identified antioxidants, the stilbene rhaponticin (isolated from F10) was the most potent (24.6±1.1 µm TE/mg). Caco-2 transport studies revealed that none of the identified compounds were detectable in basolateral samples after 2-h treatment with crude extract. In monolayers treated with F10 (60% rhaponticin), small quantities of rhaponticin were increasingly detected over time in basolateral samples with an apparent permeability coefficient (P app ) of 1.86×10 −8 cm/s (0–60 min). To model potential effects on blood redox status, we evaluated the antioxidant capacity of collected basolateral samples and observed enhanced activity over time after exposure to both extract and F10 (75 μg/mL) relative to control. Conclusions By profiling the antioxidant constituents of L. laricina bark, we identified rhaponticin as the most potent oxygen radical scavenger and observed low permeability in Caco-2 cell monolayers but an increase in basolateral antioxidant capacity.

Natural health products (NHPs), including herbal medicines, are a modality of complementary and alternative medicine (CAM) commonly used by pediatric patients with attention-deficit hyperactivity disorder (ADHD). Most families of pediatric patients find NHP treatment to be beneficial; however, clinical evidence of efficacy remains weak or lacking. Evidence of herbal medicine safety is similarly scarce, particularly with respect to herb-drug interactions and adverse events (AEs) associated with concurrent use of NHPs and ADHD prescription drugs. To support both families and physicians managing ADHD care, this review focuses on integrating available data on the safety and efficacy of herbal medicines commonly used by pediatric ADHD patients. In addition to reviewing results from clinical trials, patient surveys, and experimental studies relating to commonly used herbal medicines, the paper discusses adverse event reports involving concurrent use of herbs and ADHD drugs, identified through the FDAble database. While NHP and other CAM offer patients alternative treatment options with potential benefits as well as risks, additional research is needed to support open discussion and evidence-based decision making by families and physicians.

Objectives A novel anxiolytic natural health product (NHP) containing Souroubea sympetala and Platanus occidentalis is available for the companion animal market and is currently being developed for clinical evaluation. Addressing the risk of potential NHP–drug interactions, this study investigated S. sympetala and P. occidentalis plant extracts, and their identified bioactive compounds, for effects on the activity of cytochrome P450 (CYP) isozymes and the metabolism of the conventional anti-anxiety medication diazepam. Methods Souroubea sympetala and P. occidentalis extracts, a 1 : 1 blend of the two extracts, and five triterpenes were tested for inhibitory effects on human recombinant CYP3A4, CYP2D6, CYP2C9 and CYP2C19 activity using a fluorometric plate assay. Direct effects on the metabolism of diazepam were evaluated using human liver microsomes with drug and metabolite quantification by ultra-high-pressure liquid chromatography and mass spectroscopy. Key findings The active substances betulinic acid (BA) and ursolic acid (UA) strongly inhibited CYP3A4 activity while UA and lupeol moderately inhibited CYP2C19. All extracts exhibited strong activity against the tested isozymes at 50–100 μg/ml. BA and all plant extracts blocked the formation of major diazepam metabolites. Conclusions Betulinic acid, UA and both the extracts and blended product are expected to affect the metabolism of diazepam when given in high dose.

Wild berries, integral parts of North American Natives’ traditional nutrition, are a rich source of phytochemicals involved in the prevention of metabolic syndrome. Although phenolic compounds are generally considered to be the functional components in fruits, the identification of specific compounds in relation to this beneficial activity remains to be uncovered. With the long-term goal of establishing the contribution of phytochemical compounds to their beneficial health effects, we investigated in a comprehensive manner the composition of twelve native Canadian berries: Saskatoon berry, alpine bearberry, chokeberry, black crowberry, honeysuckle, chokecherry, cloudberry, elderberry, lowbush blueberry, alpine blueberry, lingonberry and highbush cranberry. All harvested in the province of Quebec, these fruits were analyzed for their phenolic composition (anthocyanins, flavonols, flavan-3-ols, phenolic acids, proanthocyanidins and ellagitannins) and their content in abscisic acid derivatives ( cis - and trans -ABA, abscisic acid-glucose ester, 7′-hydroxy-abscisic acid, neo -phaseic acid, phaseic acid and dihydrophaseic acid). Over 60 different phenolic compounds were identified and quantified in berries. Along with the characterization of abscisic acid, this study reports for the first time a complete characterization of proanthocyanidins in these fruits, highbush cranberry presenting the highest content of this class of compounds.

Background.Inflammation, together with related oxidative stress, is linked with the etiology of kwashiorkor, a form of severe acute malnutrition in children. A diet rich in anti-inflammatory and antioxidant phytochemicals may offer potential for the prevention and treatment of kwashiorkor. We selected and assayed five leafy green vegetables, two wild fruits, and six medicinal plants from Kenya for their antioxidant and anti-inflammatory properties. Consensus regarding medicinal plant use was established from ethnobotanical data.Methods. Antioxidant activity and phenolic content were determined using the oxygen radical absorbance capacity (ORAC) assay and Folin-Ciocalteu procedure, respectively. Anti-inflammatory activity was assessedin vitrotargeting the inflammatory mediator tumour necrosis factor-alpha (TNF-α).Results. Mangifera indica(leaves used medicinally) showed the greatest antioxidant activity (5940 ± 632 µM TE/µg) and total phenolic content (337 ± 3 mg GAE/g) butAmaranthus dubius(leafy vegetable) showed the greatest inhibition of TNF-α(IC50= 9 ± 1 μg/mL), followed byOcimum americanum(medicinal plant) (IC50= 16 ± 1 μg/mL). Informant consensus was significantly correlated with anti-inflammatory effects among active medicinal plants (r2=0.7639, P=0.0228).Conclusions. Several plant species commonly consumed by Kenyan children possess activity profiles relevant to the prevention and treatment of kwashiorkor and warrant further investigation.

There is growing evidence that fruit polyphenols exert beneficial effects on the metabolic syndrome, but the underlying mechanisms remain poorly understood. In the present study, we aimed to analyse the effects of polyphenolic extracts from five types of Arctic berries in a model of diet-induced obesity. Male C57BL/6 J mice were fed a high-fat/high-sucrose (HFHS) diet and orally treated with extracts of bog blueberry (BBE), cloudberry (CLE), crowberry (CRE), alpine bearberry (ABE), lingonberry (LGE) or vehicle (HFHS) for 8 weeks. An additional group of standard-chow-fed, vehicle-treated mice was included as a reference control for diet-induced obesity. OGTTs and insulin tolerance tests were conducted, and both plasma insulin and C-peptide were assessed throughout the OGTT. Quantitative PCR, western blot analysis and ELISAs were used to assess enterohepatic immunometabolic features. Faecal DNA was extracted and 16S rRNA gene-based analysis was used to profile the gut microbiota. Treatment with CLE, ABE and LGE, but not with BBE or CRE, prevented both fasting hyperinsulinaemia (mean ± SEM [pmol/l]: chow 67.2 ± 12.3, HFHS 153.9 ± 19.3, BBE 114.4 ± 14.3, CLE 82.5 ± 13.0, CRE 152.3 ± 24.4, ABE 90.6 ± 18.0, LGE 95.4 ± 10.5) and postprandial hyperinsulinaemia (mean ± SEM AUC [pmol/l × min]: chow 14.3 ± 1.4, HFHS 31.4 ± 3.1, BBE 27.2 ± 4.0, CLE 17.7 ± 2.2, CRE 32.6 ± 6.3, ABE 22.7 ± 18.0, LGE 23.9 ± 2.5). None of the berry extracts affected C-peptide levels or body weight gain. Levels of hepatic serine phosphorylated Akt were 1.6-, 1.5- and 1.2-fold higher with CLE, ABE and LGE treatment, respectively, and hepatic carcinoembryonic antigen-related cell adhesion molecule (CEACAM)-1 tyrosine phosphorylation was 0.6-, 0.7- and 0.9-fold increased in these mice vs vehicle-treated, HFHS-fed mice. These changes were associated with reduced liver triacylglycerol deposition, lower circulating endotoxins, alleviated hepatic and intestinal inflammation, and major gut microbial alterations (e.g. bloom of Akkermansia muciniphila, Turicibacter and Oscillibacter) in CLE-, ABE- and LGE-treated mice. Our findings reveal novel mechanisms by which polyphenolic extracts from ABE, LGE and especially CLE target the gut–liver axis to protect diet-induced obese mice against metabolic endotoxaemia, insulin resistance and hepatic steatosis, which importantly improves hepatic insulin clearance. These results support the potential benefits of these Arctic berries and their integration into health programmes to help attenuate obesity-related chronic inflammation and metabolic disorders. All raw sequences have been deposited in the public European Nucleotide Archive server under accession number PRJEB19783 ( https://www.ebi.ac.uk/ena/data/view/PRJEB19783 ).

Evidence supports the health promoting benefits of berries, particularly with regard to the prevention and management of chronic diseases such cardio- and cerebrovascular disease, diabetes and Alzheimer’s disease. Two related pathophysiological features common to many of these conditions are oxidative stress and the accumulation of advanced glycation endproducts (AGEs). Whereas antioxidant properties are well-established in several species of berries and are believed central to their protective mechanisms, few studies have investigated the effects of berries on AGE formation. Here, employing a series of complementary in vitro assays, we evaluated a collection of wild berry extracts for 1) inhibitory effects on fluorescent-AGE and Ne- (carboxymethyl)lysine-albumin adduct formation, 2) radical scavenging activity and 3) total phenolic and anthocyanin content. All samples reduced AGE formation in a concentration-dependent manner that correlated positively with each extract’s total phenolic content and, to a lesser degree, total anthocyanin content. Inhibition of AGE formation was similarly related to radical scavenging activities. Adding antiglycation activity to the list of established functional properties ascribed to berries and their phenolic metabolites, our data provide further insight into the active compounds and protective mechanisms through which berry consumption may aid in the prevention and treatment of chronic diseases associated with AGE accumulation and toxicity. As widely available, safe and nutritious foods, berries represent a promising dietary intervention worthy of further investigation.

A new anti-anxiety dietary supplement was developed for the animal health market, by combining 2 triterpene containing botanicals, Souroubea sympetala (Marcgraviaceae) with Platanus occidentalis (Platanaceae) A validated method for a quality control of the botanical blend was achieved using HPLC-APCIMS. The method resulted in the detection and quantitative determination of betulinic acid (1), and ursolic acid (2) in P. occidentalis and 1, 2, lupeol (3), β-amyrin (4) and α-amyrin (5) in S. sympetala and the finished product ZentrolTM. Detections were at low ng on column for 1 and 2 and in low µg range for 3, 4 and 5 using calibration curves within 10-100 ng (R2 > 0.993). Recovery of spiked samples for all the recoveries observed were > 94%. Inter-day and intraday variations were 0.8-3.5% and 5-10.4%, respectively. These results indicate the suitability of the developed analytical method to detect and quantify triterpenes of raw materials used in the manufacture of natural health products Se desarrolló un nuevo suplemento dietético contra la ansiedad para el mercado de la salud animal, mediante la combinación de 2 botánicos que contienen triterpenos, Souroubea sympetala (Marcgraviaceae) con Platanus occidentalis (Platanaceae) Se logró un método validado para un control de calidad de la mezcla botánica utilizando HPLC-APCIMS . El método resultó en la detección y determinación cuantitativa de ácido betulínico (1) y ácido ursólico (2) en P. occidentalis y 1, 2, lupeol (3), β-amirina (4) y α-amirina (5) en S. sympetala y el producto terminado ZentrolTM. Las detecciones fueron de ng bajo en la columna para 1 y 2 y en un rango de µg bajo para 3, 4 y 5 utilizando curvas de calibración dentro de 10-100 ng (R2 > 0,993). La recuperación de las muestras enriquecidas para todas las recuperaciones observadas fue > 94 %. Las variaciones interdiarias e intradiarias fueron 0,8-3,5% y 5-10,4%, respectivamente. Estos resultados indican la idoneidad del método analítico desarrollado para detectar y cuantificar los triterpenos de las materias primas utilizadas en la fabricación de productos naturales para la salud. University of Ottawa, Canada Universidad Nacional, Costa Rica Universidad Federal, Brasil Stonehedge Bioproducts, Canada INECOL, México Escuela de Química

Palely pigmented inflorescences of cultivated Cleome hassleriana Chodat (spider flower) have a unique two-toned appearance shown in the present study to be due to a loss of petal pigmentation within 24 h of anthesis, accompanied by an equally unique loss of petal mass. A similar loss occurs in deeply pigmented petals but is less evident to the eye because of the high initial content due to the presence, in the petal mesophyll, of globular anthocyanic vacuolar inclusions (AVIs). Inflorescences of the wild species, Cleome serrulata Pursh. (Rocky Mountain bee flower) are also two-toned because of the deeper pink colour of the unopened bud. No AVIs were seen. The pink colour of the bee flower petals is due to the same five acylated cyanidin glycosides as those previously isolated from mauve petals of spider flower. The structural pattern of the spider flower anthocyanins is shared with at least three genera of the Brassicaceae.

Obesity is an epidemic in most developed countries and novel therapeutic approaches are needed. In the course of a screening project of medicinal plants used by the Eastern James Bay Cree of Canada and having potential for the treatment of diabetes, we have identified several products that inhibit adipogenesis, suggesting potential antiobesity activities. The inhibitory activity of two of these, the extract of the inner bark of the deciduous trees Alnus incana ssp. rugosa (Speckled Alder) and Populus balsamifera L. (Balsam Poplar), was analyzed using the 3T3-L1 cell model of adipogenesis. Intracellular triglyceride accumulation, pre-adipocyte proliferation, and PPAR- γ activity were measured. Alnus incana extracts acted early in the differentiation process but did not affect clonal expansion of pre-adipocytes nor the morphological transformation from fibroblast-like to rounded fat-laden cells. Alnus incana extracts were found to act as partial agonists toward PPAR- γ activity. In contrast, Populus balsamifera extracts completely abrogated adipogenesis, severely limited clonal expansion of pre-adipocytes and generally maintained cells in an undifferentiated fibroblast-like morphology. Populus balsamifera extracts exerted antagonistic action against PPAR- γ activity. It is concluded that, through their actions on the adipocyte, these plant products may be useful for the treatment of obesity and related metabolic diseases.

Introduction – Ericaceae medicinal plants are traditionally used by the Eeyou Istchee Cree and other northern peoples of North America to treat type 2 diabetic symptoms. Because of the importance of phenolics as potential cures for degenerative diseases including type 2 diabetes, an analytical method was developed to detect them in the leaf extracts of 14 Ericaceae plants. Objective – To develop an optimised method which is applicable to a relatively large number of Ericaceae plants using their leaf extracts. For this purpose phenolics with a wide range of polarity, including a glucosylated benzoquinone, two phenolic acids, three flavanols, a flavanone, a flavone and five flavonols, were included in this study. Methodology – Characterisation of phytochemicals in extracts was undertaken by automated matching to the UV spectra to those of an in house library of plant secondary metabolites and the authentication of their identity was achieved by reversed phase-high-performance chromatography–diode array detection–atmospheric pressure chemical ionisation/mass selective detection. Results – Twenty-six phenolics were characterised within 26 min of chromatographic separation in 80% ethanol extracts of 14 Ericaceae plants. The calibration curves were linear within 0.5–880 µg/g dry mass of the plant with regression values better than 0.995. The limits of detection ranged from 0.3 for µg/mL for (+)-catechin to 2.6 µg/mL for chlorogenic acid. This is a first study dealing with relatively large number of Ericaceae extracts and is applicable to other plants of same family. Copyright © 2010 John Wiley & Sons, Ltd.

Introduction – Commercially available herbal mixture FE, a proprietary natural health product manufactured by Flora Manufacturing and Distributing Ltd (Flora), is a unique North American traditional herbal product. FE is a chemically complex mixture of eight herbs and has not been subjected to phytochemical analysis. Objective – To develop analytical methods to undertake detailed phytochemical analyses of FE, and its eight contributing herbs, including burdock (Arctium lappa L.), sheep sorrel (Rumex acetosella L.), Turkish rhubarb (Rheum palmatum L.), slippery elm Muhl. (Ulmus rubra), watercress (Nasturtium officinale R. Br.), red clover (Trifolium pratense L.), blessed thistle (Cnicus benedictus L.) and kelp (Laminaria digitata Lmx.). Methodology – The identification was undertaken by a combination of reversed phase high performance liquid chromatography–diode array detection–atmospheric pressure chemical ionisation–mass selective detection (RP-HPLC-DAD-APCI-MSD) analysis and phenolics metabolomic library matching. Results – New separation methods facilitated the identification of 43 markers in the individual herbs which constitute FE. Sixteen markers could be identified in FE originating from four contributing herbs including four caffeoyl quinic acids, three dicaffeoyl quinic acids and two caffeic acid derivatives from A. lappa, luteolin-7-O-glucoside, luteolin, five apigenin glycosides and apigenin from R. acetocella and N. officinale and sissostrin from T. pretense. A validated method for quantitative determination of three markers is reported with good intraday, interday and interoperator repeatability using a reliable alcohol based extraction technique. Conclusion – FE and its contributing herbs predominantly contain phenolics. This methodology can be applied to further develop full-scale validation of this product. Copyright © 2009 John Wiley & Sons, Ltd.

The incidence of type 2 diabetes mellitus has reached epidemic proportions worldwide. Canadian aboriginal communities, particularly the Cree Nation of Eeyou Istchee, have been identified as a high-risk population. Culturally acceptable treatment options are limited notably for diabetic complications resulting in peripheral neuropathy. Here, we describe results of an ongoing collaborative research project with Cree of Eeyou Istchee to identify botanicals capable of protecting peripheral neuronal precursors from glucose toxicity and glucose deprivation in vitro. Polar fractions of three plant organs (needles, cone, and bark) collected from Picea glauca (Moench) Voss (Pinaceae), were tested for toxicity under normoglucose, glucotoxicity, and glucose deprivation conditions. The profile of phenolic metabolites in each extract was first characterized by high-performance liquid chromatography-diode array detection-atmospheric pressure chemical ionisation mass spectrometry (HPLC-DAD-APCI/MS). We report here that these fractions are well-tolerated by PC12 neuronal precursors under normoglucose conditions. LD50 concentrations of needle extracts exceeded 100 µg/mL, whereas the LD50 of bark and cone extracts was 40 and 36.4 µg/mL, respectively. We further show that the cytoprotective properties of minhikw after glucose challenge are concentration-dependent and organ-specific. Needle

The anti-tumour activities of many plant phenolics at high concentrations (>100 μmol/L) suggest their potential use as dietary supplements in cancer chemoprevention and cancer chemotherapy. However, it is not clear what impact phenolic compounds have at the physiological concentrations obtained through consumption of high phenolic diets on neoplastic cells. In the present study, 54 naturally occurring phenolics were evaluated at physiologically relevant concentrations for their capacity to alter PC12 cell viability in response to serum deprivation, the chemotherepeutic agent etoposide, and the apoptogen C2-ceramide. Surprisingly, novel mitogenic, cytoprotective, and antiapoptotic activities were detected. Quantitative structure–activity relationship modelling indicated that many of these activities could be predicted by compound lipophilicity, steric bulk, and (or) antioxidant capacity, with the exception of inhibition of ceramide-induced apoptosis. Where quantitative structure–activity relationship analysis was insufficient, biochemical assessment demonstrated that the benzoate orsellinic acid blocked downstream caspase-12 activation following ceramide challenge. These findings demonstrate substantive mitogenic, cytoprotective, and antiapoptotic biological activities of plant phenolics on neoplastic cells at physiologically relevant dietary concentrations that should be considered in chemopreventive and chemotherapeutic strategies.

Non-enzymatic glycation and the accumulation of advanced glycation end products (AGEs) are associated with various disease states, including complications of diabetes and aging. Secondary metabolites from several plant species are known to inhibit non-enzymatic glycation and the formation of AGEs, including flavonoids found in the style (silk) of Zea mays (maize). Thirteen modern maize inbreds and one land race were tested for in vitro inhibition of non-enzymatic glycation of bovine serum albumin. Many of the tested extracts exhibited inhibitory activity, in particular the newest inbreds, which were bred for resistance to gibberella ear rot (Fusarium graminearum) and common smut (Ustilago maydis). The most active maize genotype (CO441), displaying an IC50 of 9.5 microg/mL, was more effective than aminoguanidine, a known inhibitor of glycation. Zapalote chico, a land race with high maysin content, showed only moderate inhibitory activity compared with the modern maize genotypes. Antiglycation activity was highly correlated with the total phenolic content of silk extracts and mildly correlated with resistance to certain fungal infections. The results identify modern resistant and high phenolic maize inbreds as promising candidates for the development of natural AGE inhibitors for the prevention and treatment of diabetic complications and the degenerative effects of aging.