Mining the chemical diversity of the hemp seed (Cannabis sativa L.) metabolome: discovery of a new molecular family widely distributed across hemp.

Hemp (Cannabis sativa L.) is a widely researched industrial crop with a variety of applications in the pharmaceutical, nutraceutical, food, cosmetic, textile, and materials industries. Although many of these applications are related to its chemical composition, the chemical diversity of the hemp metabolome has not been explored in detail and new metabolites with unknown properties are likely to be discovered. In the current study, we explored the chemical diversity of the hemp seed metabolome through an untargeted metabolomic study of 52 germplasm accessions to 1) identify new metabolites and 2) link the presence of biologically important molecules to specific accessions on which to focus on in future studies. Multivariate analysis of mass spectral data demonstrated large variability of the polar chemistry profile between accessions. Five main groups were annotated based on their similar metabolic fingerprints. The investigation also led to the discovery of a new compound and four structural analogues, belonging to a previously unknown chemical class in hemp seeds: cinnamic acid glycosyl sulphates. Although variability in the fatty acid profiles was not as marked as the polar components, some accessions had a higher yield of fatty acids, and variation in the ratio of linoleic acid to α-linolenic acid was also observed, with some varieties closer to 3:1 (reported as optimal for human nutrition). We found that that cinnamic acid amides and lignanamides, the main chemical classes of bioactive metabolites in hemp seed, were more concentrated in the Spanish accession Kongo Hanf (CAN58) and the French accession CAN37, while the Italian cultivar Eletta Campana (CAN48) demonstrated the greatest yield of fatty acids. Our results indicate that the high variability of bioactive and novel metabolites across the studied hemp seed accessions may influence claims associated with their commercialization and inform breeding programs in cultivar development.

The key phytochemistry of rosemary (Salvia rosmarinus) contributing to hair protection against UV.

Extracts from rosemary (Salvia Rosmarinus) are analysed for their phytochemistry using LC-MS and the phytochemistry identified. The same extracts were tested for their efficacy to act as antioxidants by both hydrogen-atom transfer (ORAC) and single electron transfer (FRAP). A correlation analysis was performed to identify the key phytochemistry responsible for antioxidant efficacy. The top performing extracts were then tested in a peptide model and in hair with the presence of UV to measure ability to protect against UV-induced peptide and protein damage. Polyphenols (e.g. rosmarinic acid, glycosides of selgin) and abietane diterpenes (e.g. carnosic acid) in rosemary were identified as the principal compounds which enables the extracts to protect hair from UV. OBJECTIVE: The objective of this work was to correlate the phytochemistry of rosemary (Salvia rosmarinus), a botanical with known antioxidant properties, to a UV protection benefit in hair. These data will give insights into mechanisms of UV damage, the ROS formed and their reactivity. METHODS: LC-MS was used to compare the compounds in 10 commercial extracts of rosemary. ORAC (oxygen radical antioxidant capacity) and FRAP (ferric reducing antioxidant power) were used to measure the antioxidant capacity of the rosemary extracts. The ORAC assay measures ability of an antioxidant to react with a peroxyl radical via hydrogen atom extraction and FRAP measures electron transfer through reduction of ferric iron (Fe3+ ) to ferrous iron (Fe2+ ) by antioxidants present in the samples. Correlation of extract composition with antioxidant measures was performed using principal component analysis. Selected extracts were assessed for their ability to protect hair from UV damage in a model peptide system and on hair. In addition, the same methods were used to test rosmarinic acid and carnosic acid, key phytochemistries in the rosemary extracts. The model system was a peptide and its decomposition on exposure to UV was monitored by LC-MS in the absence and presence of the rosemary extracts. Hair degradation in the presence of UV was measured by exposure of UV in an Atlas weatherometer followed by extraction of degraded protein in water. A fragment of the S100A3 protein was used as a marker of UV damage (m/z = 1278) and quantified via LC-MS. RESULTS: Ten rosemary extracts were assessed for antioxidant performance and correlated with their compositions. The phytochemistry in each extract varied widely with a total of 33 individual compounds identified. The differences were most likely driven by the solvent and extraction method used by the supplier with extracts varying in the proportion of polar or non-polar compounds. This did influence their reactivity in the ORAC and FRAP assays and their efficacy in preventing protein damage. Two of the key compounds identified were rosmarinic acid and carnosic acid, with rosmarinic acid dominating in extracts with mainly polar compounds and carnosic acid dominating in extracts with mainly nonpolar compounds. Extracts with higher rosmarinic acid correlated with ORAC and FRAP scores, with UV protection on hair and in the peptide model system. The extracts chosen for hair experiments showed hair protection. UV protection was also measured for rosmarinic and carnosic acid. CONCLUSIONS: Despite the variation in the profile of phytochemistries in the 10 rosemary extracts, likely driven by the chosen extraction method, all rosemary extracts had antioxidant activity measured. This study suggests that the polyphenols (e.g. rosmarinic acid, glycosides of selgin) and abietane diterpenes (e.g. carnosic acid) are the principal compounds which enables the extracts to protect hair from UV.

INTRODUCTION: Les extraits de romarin (Salvia Rosmarinus) sont analysés par LC-MS pour établir et identifier leur profil phytochimique. Les mêmes extraits ont été testés pour leur efficacité à agir comme antioxydants à la fois par transfert d'atome d'hydrogène (ORAC) et par transfert d'électrons uniques (FRAP). Une analyse de corrélation a été réalisée pour identifier les propriétés phytochimiques clés responsables de l'efficacité antioxydante. Les extraits les plus performants ont ensuite été testés dans un modèle peptidique et sur les cheveux en présences d'UV pour mesurer la capacité à protéger contre les dommages induits par les UV su les peptides et protéines. Les polyphénols (par ex. acide rosmarinique, glycosides de selgin) et les diterpènes d'abiétine (par ex. acide carnosique) dans le romarin ont été identifiés comme les principaux composés permettant aux extraits de protéger les cheveux des UV. OBJECTIF: L'objectif de ce travail était de mettre en corrélation la phytochimie du romarin (Salvia rosmarinus), une plante aux propriétés antioxydantes connues, et les bénéfices d'une protection contre les UV dans les cheveux. Ces données fourniront des informations sur les mécanismes des dommages causés par les UV, la formation du ROS et leur réactivité. MÉTHODES: La LC-MS a été utilisée pour comparer les composés de 10 extraits commerciaux de romarin. L'ORAC (Oxygen Radical Antioxidant Capacity/Capacité d'absorption des radicaux d'oxygène) et la FRAP (Ferric Reduction Antioxidant Power/Pouvoir antioxydant de réduction ferrique) ont été utilisés pour mesurer la capacité antioxydante des extraits de romarin. Le dosage ORAC mesure la capacité d'un antioxydant à réagir avec un radical peroxyl par extraction d'atome d'hydrogène et la FRAP mesure le transfert d'électrons par réduction du fer ferrique (Fe3+ ) en fer ferreux (Fe2+ ) par les antioxydants présents dans les échantillons. La corrélation entre la composition de l'extrait et les mesures des antioxydants a été effectuée en analysant les composants principaux. Les extraits sélectionnés ont été évalués pour leur capacité à protéger les cheveux des dommages causés par les UV dans un modèle de système peptidique et sur les cheveux. En outre, les mêmes méthodes ont été utilisées pour tester l'acide rosmarinique et l'acide carnosique, principales caractéristiques phytochimiques dans les extraits de romarin. Le système modèle était un peptide et sa décomposition à l'exposition aux UV a été suivie par LC-MS en l'absence et en présence des extraits de romarin. La dégradation des cheveux en présence d'UV a été mesurée par l'exposition aux UV dans un indicateur de désagrégation Atlas suivi de l'extraction de protéines dégradées dans l'eau. Un fragment de la protéine S100A3 a été utilisé comme marqueur de dommage UV (m/z = 1278) et quantifié par LC-MS. RÉSULTATS: Dix extraits de romarin ont été évalués en termes de performance antioxydante et mis en corrélation avec leurs compositions. La phytochimie de chaque extrait variait considérablement, avec un total de 33 composés individuels identifiés. Les différences étaient très probablement dues à la méthode du solvant et de l'extraction utilisée par le fournisseur avec des extraits variant dans la proportion de composés polaires ou non polaires. Cela a effectivement influencé leur réactivité dans les dosages ORAC et FRAP et leur efficacité dans la prévention des dommages protéiques. Deux des composés clés identifiés étaient l'acide rosmarinique et l'acide carnosique, l'acide rosmarinique dominant dans les extraits contenant principalement des composés polaires et l'acide carnosique dominant dans les extraits contenant principalement des composés non polaires. Les extraits avec un taux d'acide rosmarinique plus élevé étaient mis en corrélation avec les scores ORAC et FRAP, avec une protection UV sur les cheveux et dans le système de modèle peptidique. Les extraits choisis pour les expériences sur les cheveux ont montré une protection des cheveux. La protection contre les UV a également été mesurée pour l'acide rosmarinique et l'acide carnosique. CONCLUSIONS: Malgré la variation des profils phytochimiques dans les dix extraits de romarin, probablement induite par la méthode d'extraction choisie, l'activité antioxydante de tous les extraits de romarin a été mesurée. Les polyphénols (par ex. acide rosmarinique, glycosides de selgin) et les diterpènes d'abiétane (par ex. acide carnosique) dans le romarin ont été identifiés comme les principaux composés permettant aux extraits de protéger les cheveux contre les UV.

Chemical Diversity of Flavan-3-Ols in Grape Seeds: Modulating Factors and Quality Requirements.

Grape seeds are a rich source of flavan-3-ol monomers, oligomers, and polymers. The diverse profile of compounds includes mainly B-type procyanidins (especially C4→C8 linked molecules) and the key monomers, catechin, and epicatechin that are positively implicated in the 'French Paradox'. Today grape seed nutraceuticals have become a multi-million-dollar industry. This has created incentives to elucidate the variations in chemistry across cultivars, to identify signs of adulteration, and to understand the intrinsic and extrinsic factors controlling the expression of metabolites in the seeds' metabolome. This review provides a critical overview of the existing literature on grape seed chemistry. Although the biosynthetic pathways for polymeric procyanidins in seeds have not yet been explained, abiotic factors have been shown to modulate associated genes. Research of extrinsic factors has demonstrated that the control of procyanidin expression is strongly influenced, in order of importance, by genotype (species first, then variety) and environment, as claimed anecdotally. Unfortunately, research outcomes on the effects of abiotic factors have low certainty, because effects can be specific to genotype or variety, and there is limited control over physical metrics in the field. Thus, to gain a fuller understanding of the effects of abiotic factors and biosynthetic pathways, and realise potential for optimisation, a more fundamental research approach is needed. Nevertheless, the current synthesis offers insight into the selection of species or varieties according to the profile of polyphenols, as well as for optimisation of horticultural practices, with a view to produce products that contain the compounds that support health claims.

Emergence of methicillin resistance predates the clinical use of antibiotics.

The discovery of antibiotics more than 80 years ago has led to considerable improvements in human and animal health. Although antibiotic resistance in environmental bacteria is ancient, resistance in human pathogens is thought to be a modern phenomenon that is driven by the clinical use of antibiotics1. Here we show that particular lineages of methicillin-resistant Staphylococcus aureus-a notorious human pathogen-appeared in European hedgehogs in the pre-antibiotic era. Subsequently, these lineages spread within the local hedgehog populations and between hedgehogs and secondary hosts, including livestock and humans. We also demonstrate that the hedgehog dermatophyte Trichophyton erinacei produces two ß-lactam antibiotics that provide a natural selective environment in which methicillin-resistant S. aureus isolates have an advantage over susceptible isolates. Together, these results suggest that methicillin resistance emerged in the pre-antibiotic era as a co-evolutionary adaptation of S. aureus to the colonization of dermatophyte-infected hedgehogs. The evolution of clinically relevant antibiotic-resistance genes in wild animals and the connectivity of natural, agricultural and human ecosystems demonstrate that the use of a One Health approach is critical for our understanding and management of antibiotic resistance, which is one of the biggest threats to global health, food security and development.

Protection of hair from damage induced by ultraviolet irradiation using tea (Camellia sinensis) extracts.

BACKGROUND: Damage to hair by UV is relevant to most people, and for many, it is a major source of hair damage. Prevention of UV damage is of high interest to cosmetic companies. OBJECTIVES: Describe UV damage mechanisms and link these mechanisms to measurable changes in hair protein composition and color changes resulting from breakdown of yellow-colored kynurenines. Test the power of botanical antioxidants, specifically Camellia sinensis (tea) extracts to prevent this protein damage and color change. Link specific phytochemistry of extract samples to hair performance. METHODS: Camellia sinensis (tea) extracts were analyzed by LC-MS to identify the key composition chemistries. ORAC (Oxygen Radical Antioxidant Capacity) was used to measure ability of the extract to react with a peroxyl radical via a hydrogen abstraction mechanism. Hair protein structural damage was measured by quantification of a biomarker peptide that is specific to UV-induced damage and hair color changes were measured with a spectrophotometer. RESULTS: Levels of key phytochemistry in the extracts, specifically the catechins, correlated with prevention of UV-induced protein damage and prevention of color changes due to kynurenine breakdown. Extracts with higher phytochemistry levels also had higher ORAC scores indicating that they were more effective antioxidants. CONCLUSIONS: Camellia sinensis (tea) extracts can be used as effective protective treatments for hair protection but this efficacy is linked to extract concentrations of key chemistries (catechins).

Comparative Metabolomics of Reproductive Organs in the Genus Aesculus (Sapindaceae) Reveals That Immature Fruits Are a Key Organ of Procyanidin Accumulation and Bioactivity.

Fruit from A. hippocastanum L. are used commercially for chronic venous insufficiency (CVI). The isomeric mixture of pentacyclic triterpenoid saponins (ß-aescin) exert anti-inflammatory effects. Hence, research has focused on ß-aescin, yet the diversity, accumulation, and bioactivity of organ-specific secondary metabolites represent missed pharmacological opportunities. To this end, we applied an untargeted metabolomics approach by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to the chemical profiles of flowers, immature fruits, and pedicels from 40 specimens across 18 species of Aesculus. Principal component analysis (PCA), orthogonal partial least squares (OPLS-DA), and molecular networking revealed stronger chemical differences between plant organs, than between species. Flowers are rich in glycosylated flavonoids, pedicels in organic acids and flavonoid aglycones, and immature fruits in monomeric flavan-3-ols and procyanidins. Although a high diversity of flavonoids and procyanidins was observed, the relative amounts differed by plant organ. Fruit extracts demonstrated the strongest antifungal (Saccharomyces cerevisiae) and antioxidant activity, likely from the procyanidins. Overall, secondary metabolite profiles are organ-specific, and fruits accumulate antifungal and antioxidant compounds. Due to the chemical similarity between species, similar effects may be achieved between species. This creates incentives for further exploration of the entire genus, in bioprospecting for potential therapeutic leads.

Differences in diterpenoid diversity reveal new evidence for separating the genus Coleus from Plectranthus.

Covering: up to 2019The large and medicinally important tropical plant genus Plectranthus (Lamiaceae) was recently split into three separate genera on the basis of molecular and morphological evidence; Plectranthus sensu stricto, Coleus and Equilabium. We found striking differences between the diterpenoids which strongly support this taxonomic split. Coleus is characterised by abietanes oxygenated at C-14 such as royleanones, spirocoleons and acylhydroquinones, which could be useful chemotaxonomic markers to distinguish this genus from Plectranthus s.s. In contrast, the abietanes in Plectranthus s.s. lack C-14 oxygenation, but are often acylated with unusual acids. Equilabium species do not seem to produce diterpenoids. The structures of the nearly 240 abietanes so far reported from Coleus and Plectranthus and their distribution are presented. The aim of this Highlight is to provide an overview of the differences in diterpenoid diversity between these newly defined genera, which are relevant to predict which previously understudied species could hold untapped potential for their medicinal and other economic uses, and to underpin future research on how these plants have evolved to synthesise distinct abietane types.

Barbeya oleoides Leaves Extracts: In Vitro Carbohydrate Digestive Enzymes Inhibition and Phytochemical Characterization.

This study investigated the in vitro inhibitory potential of different solvent extracts of leaves of Barbeya oleoides on key enzymes related to type 2 diabetes mellitus (α-glucosidase and α-amylase) in combination with an aggregation assay (using 0.01% Triton X-100 detergent) to assess the specificity of action. The methanol extract was the most active in inhibiting α-glucosidase and α-amylase, with IC50 values of 6.67 ± 0.30 and 25.62 ± 4.12 µg/mL, respectively. However, these activities were significantly attenuated in the presence of 0.01% Triton X-100. The chemical analysis of the methanol extract was conducted utilizing a dereplication approach combing LC-ESI-MS/MS and database searching. The chemical analysis detected 27 major peaks in the negative ion mode, and 24 phenolic compounds, predominantly tannins and flavonol glycosides derivatives, were tentatively identified. Our data indicate that the enzyme inhibitory activity was probably due to aggregation-based inhibition, perhaps linked to polyphenols.

Topical and nutricosmetic products for healthy hair and dermal antiaging using "dual-acting" (2 for 1) plant-based peptides, hormones, and cannabinoids.

One of the side effects of oral antiaging retinoids is increased hair shedding. Retinoids promote the expression of TGF-ß2 from fibroblasts, which stimulate collagen expression but silences keratinocytes. Since keratinocytes normally influence differentiation of dermal papilla cells at the base of the hair follicle, retinoids feasibly inhibit hair growth via the increased expression of TGF-ß2, which inhibits Wnt/ß-catenin signaling. Fortunately, the plant kingdom provides an array of alternatives as dual-acting nutricosmetics and topicals that work independently of TGF-ß2 to confer dermal antiaging and hair health effects. These alternatives include "plant hormones" such as cytokinins and phytoestrogens. Many cytokinins are agonists of the G-coupled adenosine receptors. Partial agonism of adenosine receptors promotes collagen synthesis independently of TGF-ß2 signaling. Adenosine expression is potentially also the mechanism of minoxidil in promotion of scalp hair growth. Because of crosstalk between adenosine and cannabinoid receptors it makes sense to try combinations of specific CB2 agonists and cytokinins (or phytoestrogens). However, dual-acting cosmetics including peptides with high numbers of positively charged amino acids, such as lysine or arginine, offer real potential as they can be processed from multiple botanical candidates, including almond, fenugreek, pea sprouts, soy, and seaweeds. The current review summarizes much of what is known about retinoid alternatives in the plant kingdom and identifies potentially fruitful new areas of research.

Pharmacodynamics of Aloe vera and acemannan in therapeutic applications for skin, digestion, and immunomodulation.

Scientific studies of Aloe vera have tentatively explained therapeutic claims from a mechanistic perspective. Furthermore, in vitro outcomes demonstrate that the breakage of acemannan chains into smaller fragments enhances biological effects. These fragments can intravenously boost vaccine efficacy or entrain the immune system to attack cancer cells by mannose receptor agonism of macrophage or dendritic cells. With oral consumption, epithelialisation also occurs at injured sites in the small intestine or colon. The main advantage of dietary acemannan is the attenuation of the digestive process, increasing satiety, and slowing the release of sugars from starches. In the colon, acemannan is digested by microbes into short-chain fatty acids that are absorbed and augment the sensation of satiety and confer a host of other health benefits. In topical applications, an acemannan/chitosan combination accelerates the closure of wounds by promoting granular tissue formation, which creates a barrier between macrophages or neutrophils and the wound dressing. This causes M2 polarisation, reversal of inflammation, and acceleration of the re-epithelialisation process. This review summarises and explains the current pharmacodynamic paradigm in the context of acemannan in topical, oral, and intravenous applications. However, due to contradictory results in the literature, further research is required to provide scientific evidence to confirm or nullify these claims.

Cyanogenic Derivatives as Chemical Markers for the Authentication of Commercial Products of Bamboo Shoots.

The authentication of bamboo shoots found in the marketplace is complex because the chemical profile of processed and unprocessed material is different. During processing, heat derivatives of the potentially toxic cyanogenic glycoside taxiphyllin are produced. Here, we report the isolation and structure elucidation of the two major diarylbutenedinitrile derivatives, which are cis and trans isomers of the rare 2,3-bis(4-hydroxyphenyl)but-2-enedinitrile from a commercial extract of bamboo shoots. These compounds, absent in fresh bamboo shoots, were produced by boiling the shoots of Bambusa vulgaris and were associated with a decrease in levels of taxiphyllin. Furthermore, (E)-2,3-bis(4-hydroxyphenyl)but-2-enedinitrile was quantified in all 16 of the commercial products tested. Its abundance was found to be highly variable, ranging from 1 to 3 mg/g in preserved bamboo shoots and from 10 to 160 mg/mL in commercial aqueous extracts. Of the 15 authenticated bamboo samples tested for taxiphyllin, it was found only in the shoots of B. vulgaris and Gigantochloa verticillata, which represent two edible bamboo species. Our results indicate that diarylbutenedinitriles can be used as markers for the authentication of boil-processed bamboo shoots obtained from taxiphyllin-containing edible species and organs.

Natural flavone tricin exerted anti-inflammatory activity in macrophage via NF-κB pathway and ameliorated acute colitis in mice.

BACKGROUND: Ulcerative colitis is a subtype of inflammatory bowel disease, characterized by relapsing inflammation in the gastrointestinal tract with limited treatment options. Previous studies suggested that the natural compound tricin, a flavone isolated from rice bran, could suppress chemically-induced colitis in mice, while our recent study also demonstrated the anti-metastatic effect of tricin in colon tumor-bearing mice. HYPOTHESIS/PURPOSE: Here we further investigated the underlying mechanism of the inhibitory effects of tricin on lipopolysaccharides-activated macrophage RAW264.7 cells and explored the efficacy of tricin in acute colitis mouse model induced by 4.5% dextran sulfate sodium (DSS) for 7 days. METHODS: Tricin (75, 100, and 150 mg/kg) or the positive control drug sulfasalazine (200 mg/kg) were orally administered to mice for 7 days. Stool consistency scores, stool blood scores, and body weight were recorded daily. Disease activity index (DAI) was examined on day 7, and colon tissues were collected for biochemical analyses. The fecal microbiome of colitis mice after tricin treatment was characterized for the first time in this study using 16S rDNA amplicon sequencing. RESULTS: Results showed that tricin (50 µM) remarkably reduced nitric oxide production in lipopolysaccharides-activated RAW264.7 cells and the anti-inflammatory activity of tricin was shown to act through the NF-κB pathway. Besides, tricin treatment at 150 mg/kg significantly reversed colon length reduction, reduced myeloperoxidase activities and DAI scores, as well as restored the elevated myeloid-derived suppressive cells population in acute colitis mice. The influence from DSS on gut microbiota, such as the increased population of Proteobacteria phylum and Ruminococcaceae family, was shown to be relieved after tricin treatment. CONCLUSION: Our present study firstly demonstrated that tricin ameliorated acute colitis by improving colonic inflammation and modulating gut microbiota profile, which supports the potential therapeutic use of tricin for colitis treatment.

A Natural Flavone Tricin from Grains Can Alleviate Tumor Growth and Lung Metastasis in Colorectal Tumor Mice.

Tricin, a flavone isolated from rice bran, has been shown to be chemopreventive in a colorectal cancer (CRC) mouse model. This study aimed to illustrate the inhibitory activities of tricin in colon cancer cells and in a metastatic CRC mouse model. BALB/c mice injected with mouse Colon26-Luc cells into the rectum wall were treated with tricin (37.5 mg/kg) daily for 18 days. Orthotopic colon tumor growth and metastasis to lungs were assessed by in vivo bioluminescence imaging. Results showed that tricin suppressed Colon-Luc cells motility and downregulated phosphorylated Akt, Erk1/2 and NF-κB expressions of human colon cancer HT-29 cells. While tricin treatment suppressed tumor growth and lung metastasis as well as altered the populations of myeloid-derived suppressor cells and regulatory T cells in spleens. In summary, the tumor microenvironment modulatory and anti-metastatic effects of tricin in colon cancer mouse model were shown for the first time, suggesting the potential development of tricin-containing food supplements for CRC patients.

Antiinflammatory and Hepatoprotective Medicinal Herbs as Potential Substitutes for Bear Bile.

Practitioners of traditional Chinese medicine (TCM) commonly prescribe medicinal formulations relying on the purported synergism of a combination of plant species, sometimes incorporating animal parts and minerals. Bear bile, obtained from either wild or farmed bears, is a commonly used constituent of traditional medicine formulations. With several bear species now listed under Convention on International Trade in Endangered Species of Wild Fauna and Flora as threatened with extinction and with bear farming being actively campaigned against on ethical grounds, it is important to seek and promote alternatives to the use of bear bile as medicine. This chapter describes and evaluates the scientific data relating to the efficacy of bear bile and potential alternatives to its use, including the use of bile from other animal species, the use of synthetic chemical alternatives, and the use of herbal substitutes. Scientific studies have confirmed the efficacy of bear bile as an antiinflammatory and a hepatoprotective agent. Ursodeoxycholic acid (UDCA), the active component of bear bile is used in a synthetic form in Western medicine and can serve as an alternative to bear bile in the treatment and management of certain cholestatic liver conditions. In TCM practice, bile from domesticated animal species (such as cattle, chicken, and pig) has been used as a substitute for bear bile. Following evaluation of TCM literature and pharmacological/clinical data, the authors propose six plant species, either as single herbs or in combination, Gardenia jasminoides (zhi zi; ), Scutellaria baicalensis (huáng qín; ), Coptis chinensis (huáng lián, ), Phellodendron amurense (huáng bai; ), Andrographis paniculata (chuan xin lian; ), and Rheum palmatum (dà huang; ), two medicinal Kampo formulations, Orengedokuto, Dia-Orengedokuto (which originated from traditional Chinese herbal formula Huanglian Jiedu Tang, ), and two individual phytochemicals (berberine and andrographolide) as alternatives to bear bile. The proposed herbal alternatives are frequently found listed in traditional formulations also containing bear bile, usually with different therapeutic roles ascribed to them. The existing evidence base for the effectiveness of herbal alternatives is sufficiently strong for TCM practitioners and consumers to consider using these without the addition of bear bile. This consideration is driven by the imperative to protect populations of bears from overexploitation in the wild and when farmed. However, for the identified alternatives to be accepted by users, it is essential that researchers and TCM practitioners collaborate effectively to initiate consumer behavior change.

Medicinally Used Asarum Species: High-Resolution LC-MS Analysis of Aristolochic Acid Analogs and In vitro Toxicity Screening in HK-2 Cells.

Species of Asarum are used in traditional Chinese medicine and, similar to members of the genus Aristolochia, they contain aristolochic acid analogs (AAAs). These compounds are known for their nephrotoxic and carcinogenic effects. So far, the phytochemistry and nephrotoxicity of species of Asarum is not well studied. A high-resolution LC-MS-based metabolomic approach was used to study the phytochemical variation in medicinally used Asarum species. The cytotoxicity of the samples was assessed using human kidney (HK-2) cells. The majority of samples contained potentially nephrotoxic AAAs, including 9-methoxy aristolactam (AL) IV, AL I, and AL IV. These compounds were present in methanol as well as water extracts. AAAs were detected in all parts of the plant. The majority of the extracts were not cytotoxic to HK-2 cells at the doses tested. However, other mechanisms relating to aristolochic acid nephropathy and cancer development, such as DNA adduct formation may occur. The results of this study provide a model for assessing lesser-known plant species for toxicity.

Biochemical Profile of Heritage and Modern Apple Cultivars and Application of Machine Learning Methods To Predict Usage, Age, and Harvest Season.

The present study represents the first major attempt to characterize the biochemical profile in different tissues of a large selection of apple cultivars sourced from the United Kingdom's National Fruit Collection comprising dessert, ornamental, cider, and culinary apples. Furthermore, advanced machine learning methods were applied with the objective to identify whether the phenolic and sugar composition of an apple cultivar could be used as a biomarker fingerprint to differentiate between heritage and mainstream commercial cultivars as well as govern the separation among primary usage groups and harvest season. A prediction accuracy of >90% was achieved with the random forest method for all three models. The results highlighted the extraordinary phytochemical potency and unique profile of some heritage, cider, and ornamental apple cultivars, especially in comparison to more mainstream apple cultivars. Therefore, these findings could guide future cultivar selection on the basis of health-promoting phytochemical content.

Variation of theanine, phenolic, and methylxanthine compounds in 21 cultivars of Camellia sinensis harvested in different seasons.

This is the first study to use chemometric methods to differentiate among 21 cultivars of Camellia sinensis from China and between leaves harvested at different times of the year using 30 compounds implicated in the taste and quality of tea. Unique patterns of catechin derivatives were observed among cultivars and across harvest seasons. C. sinensis var. pubilimba (You 510) differed from the cultivars of C. sinensis var. sinensis, with higher levels of theobromine, (+)-catechin, gallocatechin, gallocatechin gallate and theasinensin B, and lower levels of (-)-epicatechin, (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG), respectively. Three cultivars of C. sinensis var. sinensis, Fuyun 7, Qiancha 7 and Zijuan contained significantly more caffeoylquinic acids than others cultivars. A Linear Discriminant Analysis model based on the abundance of 12 compounds was able to discriminate amongst all 21 tea cultivars. Harvest time impacted the abundance of EGC, theanine and afzelechin gallate.

Anti-oxidant, anti-inflammatory, analgesic and antipyretic activities of grapevine leaf extract (Vitis vinifera) in mice and identification of its active constituents by LC-MS/MS analyses.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Vitis vinifera is used in traditional medicine for diarrhea, hepatitis and stomachaches. The objective of this study was to investigate the anti-oxidant, anti-inflammatory, analgesic and antipyretic properties of the hydroalcoholic leaf extract of Vitis vinifera (EVV) on experimental models to provide scientific basis for its use. MATERIALS AND METHODS: The EVV was chemically characterized by LC-MS/MS analyses. The in vitro antioxidant activities of the EVV extract were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay and Ferric reducing antioxidant power assay (FRAP). Analgesic activity using acetic acid induced writhing and formalin test in mice, anti-inflammatory activity using carrageenan induced paw oedema and acetic acid-induced vascular permeability in mice, and antipyretic activity using Brewer's yeast induced pyrexia in rats were evaluated at 100mg/kg, 200mg/kg, and 400mg/kg doses of the extract. RESULTS: The extract (EVV) was found to contain resveratrol, quercetin, catechin, flavone, flavonols, anthocyanin, gallic acid and epicatechin. EVV produced significant dose-response anti-inflammatory activity against carrageenan-induced paw edema. EVV at dosages of 100, 200 and 400mg/kgbw significantly reduced carrageenan-induced paw edema by 34.48% (P<0.05), 36.20% (P<0.05), and 41.37% (P<0.05) at 5h after carrageenan injection, respectively. Also EVV extract reduces significantly acetic acid-induced vascular permeability in mice dose dependently. EVV (100, 200 and 400mg/kgbw) produced significant dose-response analgesic activity in the formalin test. However, the low percentage inhibition (50%) suggests that it is not a centrally acting analgesic. Extract at dosages of 100, 200 and 400mg/kg bw, p.o. significantly reduced acetic acid-induced writhing by 48.15% (p<0.05), 57.97% (p<0.05), and 68.09% (p<0.05), respectively. The extract also caused marked dose-dependent inhibition of formalin-induced pain in the second phase (p<0.05). Statistical significant reduction in rectal temperatures was observed in standard group at 21 and 23h, and in 200mg/kg and 400mg/kg doses of the extract at 23h (p<0.05) compared with the 19h. CONCLUSIONS: The results obtained indicated potential analgesic, anti-inflammatory and antipyretic effects of them hydroalcoholic leaf extract of V. vinifera observed at doses tested which support the claim for the traditional use of the plant in the treatment of various inflammatory and pain diseases.

Protoilludane, Illudane, Illudalane, and Norilludane Sesquiterpenoids from Granulobasidium vellereum.

Two new and seven known sesquiterpene compounds were isolated from an agar plate culture of Granulobasidium vellereum, isolated from a log of Ulmus sp. The two new structures were elucidated with spectroscopic methods as an illudalane derivative, granulolactone (1), and a 15-norilludane, granulodione (9). The acaricidal and insecticidal activities of the isolated compounds were examined in vitro against two major horticultural pests, the two-spotted spider mite Tetranychus urticae and the glasshouse thrips Heliothrips haemorrhoidalis, respectively.