A comprehensive account on ethnobotany, phytochemistry and pharmacological insights of genus Celtis.

The plants of Celtis L. genus have been traditionally used to cure aches, sore throats, fevers, cancer, sexually transmitted diseases, sexual weakness, diarrhea, stomach problems, amenorrhea, menstrual disorders, kidney stones, and pain. The review aims to give a comprehensive account of the current state of ethnopharmacology, phytochemistry, and biological activities of the Celtis genus, as well as to describe the potential area of future avenues. Information on the Celtis genus was obtained from internet sources such as Google Scholar, Web of Science, PubMed, ScienceDirect, and so on by using appropriate keywords, including ethnobotanical, pharmacological, pharmaceutical, bioactivity, phytochemistry, and botanical features of the Celtis genus. This review identified 14 species in the genus Celtis that have a phytopharmacological investigation, including C.africana Burm. f., C. australis L., C. occidentalis L., C. sinensis Pers., C. philippensis Blanco., C. tetrandra Roxb., C. tessmannii Rendle., C. jessoensis Koidz., C. adolfi-friderici Engl., C. iguanaea (Jacq.) Sarg., C. laevigata Wild., C. pallida Torr., C. zenkeri Engl., and C. tournefortii Lam. This genus contains many classified phytoconstituents, such as terpenoids, organic acids, flavonoids, and volatile compounds. Their extracts and pure substances have been shown to have the same anticancer, antibacterial, anti-inflammatory, antioxidant, hepatoprotective, cardioprotective, urease-inhibiting, and antidiarrheal properties as their traditional uses. In terms of current information on ethnopharmacology, phytochemicals, and pharmacological uses, the data acquired in this review could be beneficial and needed for future research. Some phytoconstituents (for instance, kaempferol, myricetin, quercetin, and eugenol) and extracts (for example, leaves, seeds, and ripe fruits extracts of C. australis) showed tremendous results in preliminary testing with promising antimicrobial, anticancer, and urease inhibitory effects. Further research and clinical investigations are needed to develop them as lead compounds and neutraceuticals, which may provide an advance over traditional medicinal systems.

Identification of hexahydrocannabinol (HHC), dihydro-iso-tetrahydrocannabinol (dihydro-iso-THC) and hexahydrocannabiphorol (HHCP) in electronic cigarette cartridge products.

PURPOSE: Since 2021, products claiming to contain hexahydrocannabinol (HHC) and hexahydrocannabiphorol (HHCP), which are tetrahydrocannabinol (THC) analogs, have been distributed via the Internet. Owing to the presence of three asymmetric carbons in their structure, HHC and HHCP have multiple stereoisomers. This study aimed to identify the actual stereoisomers of HHC and HHCP isolated from electronic cigarette cartridge products using nuclear magnetic resonance (NMR) spectroscopy. METHODS: Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-photodiode array-mass spectrometry (LC-PDA-MS) were used for the analyses of two major peaks and one minor peak in product A and two major peaks in product B. These five compounds were isolated by silica gel column chromatography, and their structures were analyzed by 1H, 13C-NMR and various two-dimensional NMR techniques, i.e., H-H correlation spectroscopy, heteronuclear multiple quantum coherence, heteronuclear multiple-bond correlation, and nuclear Overhauser effect spectroscopy. RESULTS: Three compounds isolated from product A were identified as rel-(6aR,9R,10aR)-hexahydrocannabinol (11ß-hexahydrocannabinol; 11ß-HHC), rel-(6aR,9S,10aR)-hexahydrocannabinol (11α-hexahydrocannabinol, 11α-HHC), and a minor compound (2R,5S,6R)-dihydro-iso-tetrahydrocannabinol (dihydro-iso-THC). Meanwhile, the structural isomers of the major compound isolated from product B were identified as rel-(6aR, 9R, 10aR)-hexahydrocannabiphorol (11ß-hexahydrocannabiphorol; 11ß-HHCP) and rel-(6aR, 9S, 10aR)-hexahydrocannabiphorol (11α-hexahydrocannabiphorol; 11α-HHCP). CONCLUSIONS: The presence of both 11ß-HHC and 11α-HHC in the HHC products analyzed in this study suggests that they were most likely synthesized via the reduction reaction of Δ8-THC or Δ9-THC. Dihydro-iso-THC was probably obtained as a byproduct of the synthesis of Δ8-THC or Δ9-THC from cannabidiol. Similarly, 11ß-HHCP and 11α-HHCP in the HHCP product could stem from Δ9-tetrahydrocannabiphorol.

Hemp (Cannabis sativa L., Kompolti cv.) and Hop (Humulus lupulus L., Chinook cv.) Essential Oil and Hydrolate: HS-GC-MS Chemical Investigation and Apoptotic Activity Evaluation.

In this study, essential oils (EOs) and hydrolates (Hys) from Italian hemp (Cannabis sativa L. Kompolti cv.) and hop (Humulus Lupulus L., Chinook cv.) supply chains were chemically characterized and tested to investigate their apoptotic potential for the first time. Headspace-Gas Chromatography-Mass Spectrometry (HS-GC-MS) techniques were performed to describe their volatile chemical profile, highlighting a composition rich in terpene derivatives such as monoterpenes and sesquiterpenes among which ß-myrcene, limonene, ß-caryophyllene and α-humulene were the main constituents of EOs; in contrast, linalool, cis-p-menth-2,8-dien-1-ol, terpinen-4-ol, α-terpineol, caryophyllene oxide, and τ-cadinol were found in the Hys. The cytotoxicity activity on human leukemia cells (HL60), human neuroblastoma cells (SH-SY5Y), human metastatic adenocarcinoma breast cells (MCF7), human adenocarcinoma breast cells (MDA), and normal breast epithelial cell (MCF10A) for the EOs and Hys was studied by MTT assay and cytofluorimetric analysis and scanning and transmission electron microscopy were performed to define ultrastructural changes and the mechanism of cells death for HL 60 cells. An induction of the apoptotic mechanism was evidenced for hemp and hop EOs after treatment with the corresponding EC50 dose. In addition, TEM and SEM investigations revealed typical characteristics induced by the apoptotic pathway. Therefore, thanks to the integration of the applied methodologies with the used techniques, this work provides an overview on the metabolomic profile and the apoptotic potential of hemp and hop EOs and, for the first time, also of Hys. The findings of this preliminary study confirm that the EOs and Hys from Cannabis and Humulus species are sources of bioactive molecules with multiple biological effects yet to be explored.

Chemical composition and biological activities of whole and dehulled hemp (Cannabis sativa L.) seeds.

This study aimed to determine a complete chemical composition of eight different varieties of whole hemp seeds and eight samples of commercial dehulled hemp seeds. We also evaluated the phenolic profiles and antioxidant, cytotoxic, and antimicrobial properties of hydromethanolic seed extracts. Whole hemp seeds contain much more fibre than dehulled hemp seeds, which contain more fat and protein. Sucrose and raffinose were the most abundant soluble sugars, and citric and oxalic acids were the most abundant organic acids. In the hydromethanolic hemp seed extracts, we detected the phenolic acids ferulic acid-hexoside and syringic acid. Whole hemp seed extracts exhibited better antioxidant activity than dehulled hemp seed extracts, especially in the TBARS assay. Cytotoxic activity against NCI-H460 cells was also observed. The dehulled hemp seed extracts displayed antibacterial activity, especially against Bacillus cereus, Listeria monocytogenes, and Enterococcus faecalis, and antifungal activity to a lesser extent.

Vegetative propagation of hops (Humulus lupulus L. ): Historical approach and perspectives / Propagação vegetativa do lúpulo (Humulus lupulus L. ): Abordagem histórica e perspectivas

Hop (Humulus lupulus L.) female inflorescences are important raw materials used to produce beers, cosmetics, and medicines. Vegetative propagation is the preferred way of obtaining seedlings for commercial cultivations as female plants produce more lupulin than male plants, a component of commercial interest. It can be carried out by macropropagation (stem cuttings or rhizomes) or micropropagation. This review aimed to systematize different techniques of hop vegetative propagation, with no time frame, from searches in the main academic research bases: Capes Journal Portal, Scielo, Scopus, Web of Science, Science Direct, Google Scholar, and ResearchGate. Most studies are related to micropropagation, mainly addressing different plant regulators and concentrations, as well as types of explants and culture media, strategies to produce virus-free plants, artificial lighting, and cryopreservation. Experiments with stem cuttings are more common regarding macropropagation, but factors such as size and origin of cuttings, rooting period, and the response of different cultivars need to be better evaluated. Cultivation by cuttings allows the production of clones of female plants and micropropagation the production of virus-free clones in a short time and less physical space. Currently, micropropagation has been widely applied to cryopreservation.(AU)

As inflorescências femininas do lúpulo (Humulus lupulus L.) são matérias-primas importantes utilizadas na produção de cervejas, cosméticos e medicamentos. Como as plantas femininas produzem mais lupulina que as masculinas, componente de interesse comercial, a propagação vegetativa é a forma preferencial de obtenção de mudas para os cultivos comerciais. Esta pode ser realizada por macropropagação (estaquia caulinar ou rizomas) ou micropropagação. O objetivo desta revisão foi sistematizar as diferentes técnicas de propagação vegetativa do lúpulo, sem recorte temporal, a partir de buscas nas principais bases de pesquisa acadêmica: Portal de Periódicos Capes, Scielo, Scopus, Web of Science, Science Direct, Google Acadêmico e Research Gate. A maioria dos trabalhos são relacionados à micropropagação, abordando principalmente diferentes reguladores vegetais e concentrações, além de tipos de explantes e meios de cultura, estratégias para produzir plantas livres de vírus, iluminação artificial e criopreservação. Quanto à macropropagação, experimentos com estaquia caulinar são mais comuns, porém fatores precisam ser melhor avaliados tais como tamanho e origem das estacas, período de enraizamento e resposta de diferentes cultivares. O cultivo por estacas permite a produção de clones de plantas femininas e a micropropagação a produção de clones isentos de vírus, em pouco tempo e em menor espaço físico. Atualmente, a micropropagação tem sido muito aplicada à criopreservação.(AU)

Metabolomic Analysis of Cannabinoid and Essential Oil Profiles in Different Hemp (Cannabis sativa L.) Phenotypes.

Hemp (Cannabis sativa L.) cannabinoids and terpenoids have therapeutic effects on human and animal health. Cannabis plants can often have a relatively high heterogeneity, which leads to different phenotypes that have different chemical profiles despite being from the same variety. Little information exists about cannabinoid and terpenoid profiles in different hemp phenotypes within the same variety. For this study, 11 phenotypes from three different varieties ("Carmagnola" selected (CS), "Tiborszallasi" (TS), and "Finola" selection (FS)) were analyzed. The components of essential oil (29) were analyzed using gas chromatography with flame ionization detection (GC/FID), and 10 different cannabinoids of each phenotype were determined using high-performance liquid chromatography (HPLC). Principal component analysis (PCA) and analysis of variance (ANOVA) showed that according to the components of essential oil, FS and TS plants were more uniform than CS plants, where there were great differences between CI and CII phenotypes. The content of cannabinoid CBD-A was the highest in all four FS phenotypes. By comparing cannabinoid profiles, FS was clearly separated from TS and CS, while these two varieties were not clearly distinguishable. Phenotypes TV and CI had the highest total content of Δ-9-THC, while all phenotypes of FS had the highest total content of CBD. The highest total content of CBG was determined in phenotype CI. Obtained results are useful for the development of new supplementary ingredients, for different pharmacy treatments, and for further breeding purposes.

Bioactive spirans and other constituents from the leaves of Cannabis sativa f. sativa.

In this paper, 17 compounds (1-17) were isolated from the leaves of Hemp (Cannabis sativa f. sativa). Among the isolates, two were determined to be new spirans: cannabispirketal (1), and α-cannabispiranol 4'-O-ß-D-glucopyranose (2) by 1D and 2D NMR spectroscopy, LC-MS, and HRESIMS. The known compounds 7, 8, 10, 13, 15, and 16 were isolated from Hemp (C. sativa f. sativa) for the first time. Furthermore, compounds 8 and 13 were isolated from the nature for the first time. All isolated compounds were evaluated for cytotoxicity on different tissue-derived passage cancer cell lines through cell viability and apoptosis assay. Among these compounds, compounds 5, 9 and 16 exhibited a broad-spectrum antitumor effect via inhibiting cell proliferation and promoting apoptosis. These results obtained have provided valuable clues to the understanding of the cytotoxic profile for these isolated compounds from Hemp (C. sativa f. sativa).

Novos aspectos sobre a intoxicação por Trema micrantha (Cannabaceae) em equídeos / New aspects of Trema micrantha (Cannabaceae) poisoning in equids

Trema micrantha é uma planta arbórea distribuída amplamente no Brasil. Descrevem-se nesse trabalho novos aspectos epidemiológicos e patológicos relacionados com a intoxicação por essa planta em equídeos. Dois equinos adultos da raça Crioula e dois asininos, de localidades distintas, foram intoxicados naturalmente por Trema micrantha, após consumirem grande quantidade da planta que ficou disponível a eles após uma poda. Além disso, um cavalo adulto, de outra propriedade, que estava passando por restrição alimentar, se intoxicou após consumir as partes baixas de um grande número de árvores jovens de T. micrantha. Clinicamente, em todos os equídeos a doença se caracterizou por alterações neurológicas, que apareceram três a quatro dias após o consumo da planta, com curso clínico de dois a quatro dias (abreviadas por eutanásia). Os principais achados de necropsia foram observados no fígado, que estavam levemente aumentados e com padrão lobular evidente e no sistema nervoso central (SNC), com múltiplas áreas amareladas, focos de malacia e hemorragia, principalmente, no tronco cerebral e cerebelo. Histologicamente, havia necrose hepática aguda, edema acentuado no SNC com degeneração fibrinoide da parede de vasos associada a hemorragia e trombose e, frequentemente, com infiltrado de neutrófilos. Outras alterações observadas nos encéfalos foram: grande quantidade de astrócitos de Alzheimer tipo II na substância cinzenta, acúmulos de células Gitter e degeneração Walleriana próxima a focos de lesões vasculares.

Trema micrantha is a tree widely distributed in Brazil. This paper describes new epidemiological and pathological aspects of this plant poisoning in equids. Two adult horses, Criollo breed and two donkeys from different locations were naturally poisoned by Trema micrantha after consuming large amounts of the plant which was available to them due to pruning. Also an adult horse from another ranch, which was undergoing dietary restriction, was poisoned after consuming the lower parts of a large number of young T. micrantha trees. Clinically, in all cases, the disease was characterized by neurological symptoms that started 3 to 4 days after consumption of the plant, with a clinical course that varied 2 to 4 days (abbreviated by euthanasia). The main gross findings were in the liver, which had slightly increased lobular pattern evident, and in the central nervous system (CNS) with multiple yellowish areas, foci of malacia and hemorrhage, especially in the brainstem and cerebellum. Histologically, there was acute liver necrosis with marked edema of the CNS with fibrinoid vessel wall degeneration, hemorrhage and thrombosis as well as neutrophil infiltration. Other changes in the brain were Alzheimer type II astrocytes in the gray matter, accumulation of Gitter cells and Wallerian degeneration near to vascular lesion foci.

Intoxicação experimental por Trema micrantha (Cannabaceae) em equinos / Experimental Trema micrantha (Cannabaceae) poisoning in horses

O objetivo desse estudo foi confirmar a toxidez e caracterizar os aspectos clínicos e patológicos da intoxicação por Trema micrantha em equinos. Três equinos, pôneis, com idade entre 2 e 7 anos consumiram espontaneamente folhas de T. micrantha em doses únicas de 30g/kg, 25g/ kg e 20g/kg. Os três animais adoeceram e evoluíram para morte. Outro equino recebeu 15 e 25g/kg da planta com intervalo de 30 dias entre as doses e não apresentou alteração clínica. Coletas diárias de sangue foram realizadas para análises bioquímicas. Os principais sinais clínicos apresentados foram apatia, desequilíbrios, dificuldade de deglutição, decúbito esternal, decúbito lateral, movimentos de pedalagem, coma e morte. Os três equinos afetados apresentaram elevação da atividade sérica de gama-glutamil transferase, dos níveis séricos de amônia e diminuição da glicemia. Esses animais foram necropsiados e fragmentos de diversos órgãos foram coletados para análise histopatológica e imuno-histoquímica. Os principais achados patológicos foram encontrados no fígado e no encéfalo dos três animais. O fígado apresentava, macroscopicamente, acentuação do padrão lobular; enquanto que, no encéfalo havia áreas amareladas na superfície de corte, mais evidentes na substância branca do cerebelo. Microscopicamente, o fígado apresentava tumefação hepatocelular, necrose de coagulação predominantemente centrolobular e hemorragia associada. No encéfalo, havia edema perivascular generalizado e astrócitos Alzheimer tipo II na substância cinzenta. Esses astrócitos apresentaram marcação fraca ou negativa na imuno-histoquímica anti-GFAP e marcação positiva do antígeno S-100. A dose letal mínima de folhas de T. micrantha estabelecida nesse experimento foi de 20g/kg. A ampla distribuição e palatabilidade desta planta, associadas à alta sensibilidade da espécie equina, constatada nesse experimento, reforçam a importância da planta em casos acidentais de intoxicação em equinos.

The aim of this study was to confirm the toxicity and characterize the clinical and pathological aspects of poisoning by Trema micrantha in horses. The green leaves of the plant were spontaneously consumed by three ponies at single doses of 20, 25, and 30g/kg. The three ponies developed clinical disease characterized by apathy, equilibrium deficit, deglutition difficulty, sternal or lateral recumbency, paddling, coma and death. Affected ponies had also enhanced seric activity of gamma-glutamyl transferase, seric ammonia apart of diminished glycemia. An additional pony, that received 15 and 25g/kg of T. micrantha with a 30 day interval between the two doses, showed no clinical disease. The three ponies were necropsied and fragments from organs were collected for histology and immunostaining (IS) procedures. The main pathological findings were in the liver and encephalon. There were enhanced lobular pattern of the liver and yellowish areas in the cut surface of the encephalon, especially visualized in the cerebellar white matter. Microscopically, there was hepatocellular swelling and coagulative hepatic necrosis predominantly centrilobular with hemorrhages. Generalized perivascular edema and Alzheimer type II astrocytes were observed in the encephalon. The Alzheimer type II astrocytes showed weak or absent anti-glial fibrillar acid protein IS associated with positive IS for S-100 protein. The minimal lethal dose of Trema micrantha leaves was established at 20g/kg. The sensibility of horses to this plant, its wide distribution and high palatability reinforces the importance of Trema micrantha in accidental poisoning of horses.