ß-sitosterol isolated from the leaves of Trema orientalis (Cannabaceae) promotes viability and proliferation of BF-2 cells.

Trema orientalis is a pioneer species in the cannabis family (Cannabaceae) that is widely distributed in Thai community forests and forest edges. The mature leaves are predominantly used as an anti-parasite treatment and feed for local freshwater fish, inspiring investigation of their phytochemical composition and bioactivity. The purpose of this work was to investigate the bioactive compounds in T. orientalis leaf extract and their cytotoxicity in the BF-2 fish cell line (ATCC CCL-91). Flash column chromatography was used to produce 25 mL fractions with a mixture solvent system comprised of hexane, diethyl ether, methanol, and acetone. All fractions were profiled with HPLC-DAD (mobile phase methanol:aqueous buffer, 60:40 v/v) and UV detection (wavelengths 256 and 365 nm). After drying, a yellowish powder was isolated from lipophilic leaf extract with a yield of 280 µg/g dry weight. Structure elucidation by nuclear magnetic resonance (NMR) indicated it to consist of pure ß-sitosterol. The lipophilic extract and pure compound were evaluated for cytotoxicity using BF-2 cells. MTT assays showed both leaf extract and pure compound at 1 µg/mL to increase cell viability after 24 h treatment. The respective half maximal inhibitory concentration (IC50) values of leaf extract and ß-sitosterol were 7,027.13 and 86.42 µg/ml, indicating a lack of toxicity in the BF-2 cell line. Hence, T. orientalis can serve as a source of non-toxic natural lipophilic compounds that can be useful as bioactive ingredients in supplement feed development.

The putative cannabinoid-secreting trichome of Trema micrantha (L.) Blume (Cannabaceae).

Trema, a genus of the popularly known Cannabaceae, has recently been the subject of cannabinoid bioprospection. T. micrantha is a tree with pharmacological potential widely used in folk medicine. It has two types of glandular trichomes, bulbous and filiform, spread throughout the plant body. Considering the proximity of this species to Cannabis sativa and Trema orientalis, species containing cannabinoids, the glandular trichomes of T. micrantha are also expected to be related to the secretion of these compounds. Thus, this study aims to detail the morphology of secretory trichomes during the synthesis, storing and release of metabolites in T. micrantha. We tested the proposition that they could be a putative type of cannabinoid-secreting gland. Pistillate and staminate flowers and leaves were collected and processed for ontogenic, histochemical, and ultrastructural analyses. Both types of glandular trichomes originate from a protodermal cell. They are putative cannabinoid-secreting sites because: (1) terpene-phenols and, more specifically, cannabinoids were detected in situ; (2) their secretory subcellular apparatus is consistent with that found in C. sativa: modified plastids, polyribosomes, an extensive rough endoplasmic reticulum, and a moniliform smooth endoplasmic reticulum. Plastids and smooth endoplasmic reticulum are involved in the synthesis of terpenes, while the rough endoplasmic reticulum acts in the phenolic synthesis. These substances cross the plasma membrane by exocytosis and are released outside the trichome through cuticle pores. The study of the cell biology of the putative cannabinoid glands can promote the advancement of prospecting for natural products in plants.

Impact of copper-based fungicides on the antioxidant quality of ethanolic hop extracts.

Hops contain a variety of compounds possessing antioxidant capacity including phenolic and polyphenolic compounds as well as α- and ß- acids. These compounds may contribute to the oxidative stability of beer during brewing and storage. Hop plants may be treated with copper-based fungicides (CBFs) which have been shown to increase the total copper content of harvested hop cones; however, copper ions are well known to catalyze the generation of reactive oxygen species production in beer and may negatively impact its oxidative stability. Increased copper content in CBF-treated hops has been previously shown to have deleterious effects on the aroma quality of hops and beer. The impact of CBFs on the antioxidant content and quality of hops has not been previously investigated. In this study, ethanolic extracts of CBF-treated hops are evaluated for their metal content and antioxidant quality in order to determine whether excess copper from CBF treatment negatively impacts their antioxidant capacity.

Chemical characterization, pharmacological effects, and toxicity of an ethanol extract of Celtis pallida Torr. (Cannabaceae) aerial parts.

ETHNOPHARMACOLOGICAL RELEVANCE: Celtis pallida Torr (Cannabaceae) is employed as a folk medicine for the treatment of inflammation, pain, skin infections, and diarrhea, among other diseases. AIM OF THE STUDY: The purpose of this work was to assess the chemical composition, the in vitro and in vivo toxicity, the antimicrobial, anti-inflammatory, antidiarrheal, antinociceptive, locomotor, and sedative effects of an ethanolic extract obtained from Celtis pallida aerial parts (CPE). MATERIALS AND METHODS: The composition of CPE was carried out by GC-MS. The in vitro and in vivo toxic activity of CPE was estimated with the comet assay (10-1000 µg/ml) for 5 h in peripheral blood mononuclear cells, and the acute toxicity test (500-5000 mg/kg p.o.), for 14 days, respectively. The antimicrobial effect of CPE was evaluated using the minimum inhibitory concentration (MIC) assay, whereas the antidiarrheal activity (10-200 mg/kg p.o.) was calculated using the castor oil test. The antinociceptive effects of CPE (50-200 mg/kg p.o.) were estimated with the acetic acid and formalin tests, as well as the hot plate test. The sedative and locomotor activities of CPE (50-200 mg/kg p.o.) were assessed with the pentobarbital-induced sleeping time test and the rotarod test, respectively. RESULTS: The main compound found in CPE was the triterpene ursolic acid (22% of the extract). CPE at concentrations of 100 µg/ml or higher induced genotoxicity in vitro and showed low in vivo toxicity (LD50 > 5000 mg/kg p.o.). Additionally, CPE lacked (MIC > 400 µg/ml) antimicrobial activity but exerts antinociceptive (ED50 = 12.5 ±â€¯1.5 mg/kg) and antidiarrheal effects (ED50 = 2.8 mg/kg), without inducing sedative effects or altering the locomotor activity. The antinociceptive activity of CPE suggests the participation of adrenoceptors, as well as the nitric oxide/cyclic guanosine monophosphate (cGMP) pathway. CONCLUSION: C. pallida exerts its antinociceptive effects probably mediated by the nitric oxide/cyclic guanosine monophosphate (cGMP) pathway.

Assessment of antimutagenic action of Celtis glabrata Steven ex Planch. (Cannabaceae) extracts against base pair exchange and frame shift mutations on Salmonella typhimurium TA98 and TA100 strains by Ames test.

CONTEXT: Celtis glabrata is used in Turkey for the treatment of various health disorders. OBJECTIVE: The acetone, chloroform, ethanol, and methanol extracts of C. glabrata leaf, fruit, and seed were investigated to evaluate their antimutagenic activities. MATERIAL AND METHODS: The antimutagenicity of these extracts was determined by Ames test against mutagens (4-nitro-O-phenylenediamine, 2-aminofluorene (2-AF), and sodium azide (SA)). The extracts were used at concentrations between 5 and 0.005 mg/plate. RESULTS: The ethanol extracts of leaves exhibited strong antimutagenicity (70%) against 2-AF with S9 at 5 mg/plate on TA98. But methanol (61%, 53%) and acetone (53%, 52%) also revealed strong inhibition rates at concentrations of ≥ 0.5 mg/plate. Among the extracts, the highest activity (96%) was obtained from acetone extract against SA without S9, followed by chloroform extract (91%) at a dose of 5 mg/plate on TA100 with S9. Ethanol (without S9) and chloroform (with S9) extracts showed strong antimutagenicity at all doses. Exception of chloroform and acetone (without S9), all fruit extracts (with/without S9) manifested strong antimutagenicity at doses of ≥ 0.5 mg/plate on TA98 strain. Ethanol extracts revealed 68% inhibition against 2-AF on TA98. Acetone and ethanol extracts manifested 84% and 82% inhibition against SA on TA100, respectively. All the extracts of seeds revealed strong inhibition against 2-AF at ≥ 0.5 mg/plate doses on TA98, but acetone extract showed excellent antimutagenicity (94%). Moreover, the chloroform (74, 73, 63, 54%), acetone (74, 72, 70, 65%) and methanol (74, 67, 63, 61%) extracts of seeds revealed strong antimutagenic activity on TA100 against SA with S9. DISCUSSION AND CONCLUSION: This plant may be natural source of antimutagenic agents.

Acute oral toxicity of Celtis iguanaea (Jacq. ) Sargent leaf extract (Ulmaceae) in rats and mice / Toxicidade oral aguda do extrato de folhas de Celtis iguanaea (Jacq. ) Sargent em ratos e camundongos

RESUMO Celtis iguanaea (Jacq.) Sargent is popularly used to treat urinary infections, kidneys, breast, body aches, rheumatism, asthma, cramps, poor digestion and as a diuretic medicine. This study aims to determine the acute toxicity of the aqueous leaf extract of Celtis iguanaea (Jacq.) Sargent in rodents. After the collection processes, identification, drying and grinding, the lyophilized powder of the leaves produced, by infusion, the aqueous extract and it was dissolved in saline 0.9%. The administration was made by gavage at a dose of 2000 mg kg-1to rats and mice of both genders. The oral toxicity was determined according to the OECD 423 guide. Signs of toxicity were observed for 15 days and classified from 0 to 4 respectively as missing, rare, mild, moderate and severe. The weight of the animals and the physiological parameters such as food intake and excrements production were observed. All animal tissue samples were collected for histological analysis. The extract was included in Type 5 (substance with LD50 higher than 2000 mg kg-1 and less than 5000 mg kg-1), being considered of low toxicity, but the histopathologycal findings suggested nephrotoxicity and cardiotoxicity. The absolute weight of the kidneys and the heart of the male rats and mice increased, but there was no significant raise in the relative weight of the animals’ organs.

RESUMO Celtis iguanaea (Jacq.) Sargent é uma planta usada popularmente para tratar infecções do trato urinário, rim, mama, dores no corpo, reumatismo, asma, cólicas, má digestão e também é usada como diurético. Este trabalho objetivou determinar a toxicidade aguda do extrato aquoso de folhas de Celtis iguanaea (Jacq.) Sargent em roedores. Após os processos de coleta, identificação, secagem e moagem, o pó liofilizado das folhas da planta foi utilizado para produzir o seu extrato aquoso por infusão e então dissolvido em solução salina a 0.9 %. A administração foi feita por gavagem na dose de 2000 mg kg-1 em ratos e camundongos de ambos os sexos. A toxicidade oral foi determinada de acordo com o guia 423 da OECD. Sinais de toxicidade foram observados por 15 dias e tabulados de 0 a 4, respectivamente, como ausentes, raros, leves, moderados e graves. Foi acompanhado o peso dos animais e parâmetros fisiológicos tais como alimentação e excreções. Amostras do tecido de todo o animal foram coletadas para análise histológica. A toxicidade encontrada para o extrato foi incluída na classe 5 (substâncias com DL50 superior a 2000 mg kg-1 e menor que 5000 mg kg-1) sendo considerada baixa, porém, as observações histopatológicas sugerem nefrotoxicidade e cardiotoxicidade. O peso absoluto dos rins e coração de ratos e camundongos machos aumentou, porém, não houve aumento significativo no peso relativo dos órgãos dos animais.

Anti-ulcerogenic and antisecretory effects of Celtis iguanaea (Jacq. ) Sargent hexane leaf extract / Efeito anti-ulcerogênico e antisecretório do extrato hexânico das folhas de Celtis iguanaea (Jacq. ) Sargent

The Celtis iguanaea (Jacq.) Sargent (Cannabaceae) is one of the native species of the Cerrado region of Brazil widely used in folk medicine to treat dyspepsia. The objective of the present study was to evaluate the gastroprotective effect of the Celtis iguanaea (Jacq.) Sargent (HE) hexane leaf extract in the lesion and gastric secretion models.Antiulcerogenic activity of the Celtis iguanaea (HE) hexane leaf extract was observed with the experimental models, such as indomethacin and pyloric ligation-induced gastric ulcers. In order to evaluate the antisecretory activity of this extract, isolated Rana catesbeiana mucosa and pyloric ligation in mice were used. The HE treatment reduced the lesion index of indomethacin and pyloric ligation-induced ulcer. This extract also reduced the gastric acid secretion and total acidity (increasing the gastric pH) in mice. The secretion of H+ was reduced in the basal values (15.58 ± 1.99 µEq H+/g/15 min) when isolated Rana catesbeiana mucosa was incubated with HE. Intraduodenal administration of HE reduced the gastric secretion produced by bethanecol or histamine. The antiulcerogenic and antisecretory efficacy of HE in this study suggest anticholinergic and antihistaminergic mechanism or interruption of intracellular events that are linked to acid secretion.

Celtis iguanaea (Jacq.) Sargent (Cannabaceae) é uma das espécies nativas do cerrado brasileiro, sendo amplamente utilizada na medicina popular para o tratamento de dispepsia. O objetivo do presente trabalho foi avaliar o efeito gastroprotetor do extrato hexânico das folhas de Celtis iguanaea (Jacq.) Sargent (EH) em modelos de secreção e de lesões gástricas. A atividade antisecretória do extrato hexânico das folhas de Celtis iguanaea (EH) foi avaliada no modelo de mucosa isolada de Rana catesbeiana e pelo modelo de ligadura pilórica em camundongos. A avaliação da atividade anti-ulcerogênica do EH foi determinada no modelo de úlceras gástricas induzidas por indometacina e ligadura pilórica. Quando a mucosa de Rana catesbeiana isolada foi incubada com o EH, a secreção de H+ foi reduzida em comparação com o valor basal (15,58 ± 1,99 µEq H+/g/15 min). O tratamento com o EH reduziu o índice de lesões na úlcera induzida por indometacina e ligadura pilórica. O EH também reduziu o volume de secreção, a acidez total, e aumentou o pH gástrico. A administração intraduodenal reduziu a secreção gástrica produzida por betanecol ou histamina. A eficácia anti-ulcerogênica e antisecretória do EH neste estudo, sugere um mecanismo anticolinérgico e antihistaminérgico ou interrupção de eventos intracelulares que estão ligados à secreção ácida.

Bioactive phenolic amides from Celtis africana.

Nine compounds have been isolated for the first time from Celtis africana, namely trans-N-coumaroyltyramine (1), trans-N-feruloyltyramine (2), trans-N-caffeoyltyramine (3), lauric acid (4), oleic acid (5), palmitic acid (6), lupeol (7), ß-sitosterol (8) and oleanolic acid (9), respectively. Their structures have been elucidated by different spectroscopic techniques. The isolated compounds were screened for their antioxidant, anti-inflammatory and acetylcholinestrease enzyme inhibitory activities. Compounds 1-3 showed significant antioxidant and anti-inflammatory activities and weak to moderate acetylcholinestrease enzyme inhibition activity.

Cannabaceae pollen in the atmosphere of Brianza, Northen Italy.

AIM: We have noted several patients suffering for rhinitis and/or asthma symptoms in the summer months without positive skin tests for pollens blossoming in the corresponding season. So we tried to determine the pollination pattern of pollens usually not tested, because no company produces the corrisponding extract. METHODS: We determined airborne pollination by means of a volumetric pollen trap for five years from 2001 to 2005. RESULTS: We found, in addition to well-known aeroallergens, a significant percentage of Cannabaceae from the end of July to mid September with the highest concentration in 2004 (140 gr/m3). DISCUSSION: The potential allergenicity of Cannabis pollen was supported in the United States, where they found some patients with rhinitis or asthma during the pollen seasons and positive prick test for Cannabis, but the clinical significance of Cannabaceae pollen as an aeroallergen in Europe is still undefined.

Cannabis use in sickle cell disease: a questionnaire study.

Cannabinoids are increasingly being considered for the management of various painful conditions, and could be considered as an option for treating acute pain in sickle cell disease (SCD). The objective of this study was to determine the extent of use of cannabis in the community for pain and other symptom relief, and its side effects during self-administration in patients with SCD. Patients attending Central Middlesex Hospital in London were invited to complete a structured self-administered anonymous questionnaire. Eighty-six young adults with HbSS, HbSC and HbSbetathalassaemia disease (median age 30 years) participated in the study. Results showed that 31 (36%) had used cannabis in the previous 12 months to relieve symptoms associated with SCD. The main route in all but two patients was by smoking. The main reasons for use were to reduce pain in 52%, and to induce relaxation or relieve anxiety and depression in 39%. Symptoms related to sedation and mood effects were reported in 77% of patients. The majority of patients (58%) expressed their willingness to participate in studies of cannabis as a medicine. We conclude that research in the use of cannabinoids for pain relief in SCD would be both important and acceptable to adult patients.