Cannabis Sativa Extract Research Articles

Virtual screening and in vitro experiments highlight cannabidiol as a drug-like phosphodiesterase 9 inhibitor.

The growing interest on the therapeutic potential against neurodegeneration of Cannabis sativa extracts, and of phytocannabinoids in particular, is paralleled by a limited understanding of the undergoing biochemical pathways in which these natural compounds may be involved. Computational tools are nowadays commonly enrolled in the drug discovery workflow and can guide the investigation of macromolecular targets for such molecules. In this contribution, in silico techniques have been applied to the study of C. sativa constituents at various extents, and a total of seven phytocannabinoids and four terpenes were considered. On the side of ligand-based virtual screening, physico-chemical descriptors were computed and evaluated, highlighting the phytocannabinoids possessing suitable drug-like properties to potentially target the central nervous system. Our previous findings and literature data prompted us to investigate the interaction of these molecules with phosphodiesterases (PDEs), a family of enzymes being studied for the development of therapeutic agents against neurodegeneration. Among the compounds, structure-based techniques such as docking and molecular dynamics (MD), highlighted cannabidiol (CBD) as a potential and selective PDE9 ligand, since a promising calculated binding energy value (-9.1 kcal/mol) and a stable interaction in the MD simulation timeframe were predicted. Additionally, PDE9 inhibition assay confirmed the computational results, and showed that CBD inhibits the enzyme in the nanomolar range in vitro, paving the way for further development of this phytocannabinoid as a therapeutic option against neurodegeneration.

Phytochemical study using HPLC-UV/GC–MS of different of Cannabis sativa L seeds extracts from Morocco

Within the framework of the valorization of cannabis seeds harvested in Ketama located in the region of Tangier-Tetouan-Al Hoceima, Morocco, a phytochemical study was carried out. The hexane extract of cannabis seeds was prepared and analyzed by gas chromatography-mass spectroscopy (GC–MS). The study proved that the natural extracts of Cannabis Sativa L seeds contain a variety of phenolic compounds to which various biological activities are attributed. Therefore, we have chosen to evaluate the quantitative estimation of total polyphenols (the Folin-Ciocalteu method) and subsequently the chemical composition of the different organic extracts by HPLC-UV. The quantitative analyses of total polyphenols determined that ethanolic and aqueous extracts were the richest with polyphenols compound. The results obtained by GC–MS reveal the presence of linoleic acid (omega 6) and 7- octadecenoic acid as main composition; first comes linoleic acid (omega 6) with 42.92% portion, followed by 7- octadecenoic acid with 22.91% portion respectively. Moreover, the analysis of the organic extracts by high-performance liquid chromatography revealed the probable presence of catechin di-hydrate, 6 hydroxy flavone, ferulic acid, 8 methoxy flavone, hesperidin acid, rutin and cinnamic acid in the dichloromethane extract, naringin acid was the main component in the ethanolic extract with a percentage of 41. 92% followed by rutin, hesperidin acid, O-dianisidine, ferulic acid, chlorogenic acid, coumarin, and cinnamic acid. On the other hand, aqueous extract was found to be rich in catechin di-hydrate, O. dianisidine, cinnamic acid, p-coumaric acid, caffeic acid, benzoic acid, and sinapinic acid.

Supercritical Extract of Cannabis sativa Inhibits Lung Metastasis in Colorectal Cancer Cells by Increasing AMPK and MAPKs-Mediated Apoptosis and Cell Cycle Arrest.

Colorectal cancer (CRC) is one of the diseases with the highest rates of prevalence and mortality despite therapeutic methods in the world. In particular, there are not enough methods to treat metastasis of CRC cells to distant organs. Cannabis sativa Linne (C. sativa) is a popular medicinal plant used by humans to treat many diseases. Recently, extracts of C. sativa have shown diverse pharmacological effects as a result of choosing different extraction methods. In this study, we performed experiments to confirm the inhibitory effect and related mechanisms of supercritical extract of C. sativa on metastatic CRC cells. The effect of SEC on the viability of CRC cell lines, CT26 and HCT116, was determined using CCK reagent. Flow cytometry was performed to confirm whether SEC can promote cell cycle arrest and apoptosis. Additionally, SEC reduced proliferation of CT26 and HCT116 cells without causing toxicity to normal colon cell line CCD-18Co cells. SEC treatment reduced colony formation in both CRC cell lines, promoted G0/G1 phase arrest and apoptosis in CT26 and HCT116 cells through AMPK activation and MAPKs such as ERK, JNK, and p38 inactivation. Moreover, oral administration of SEC decreased pulmonary metastasis of CT26 cells. Our research demonstrates the inhibitory effect of SEC on CRC cell proliferation and metastasis. Thus, SEC might have therapeutic potential for CRC treatment.

Antibacterial activities and biosynthesis of nanoparticles using hemp extracts

Background: The use of plants in the biosynthesis of nanoparticles is a fast-growing technique and has gained much interest from researchers over the years.Aim: This study reported the utilisation of leaf extracts of Cannabis sativa L. (hemp) for the biosynthesis of silver nanoparticles (AgNPs).Methods: In this study, techniques such as transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) microanalysis and Fourier transform infrared (FT-IR) were carried out for the synthesis and characterisation of nanoparticles. The antibacterial efficacy of the synthesised nanoparticles was evaluated by using the agar diffusion and macrobroth dilution methods. The antibacterial properties of the biosynthesised AgNPs were evaluated against both Gram-negative and Gram-positive bacteria.Results: The formation of AgNPs was confirmed by colour change in plant extracts and further characterised by Ultraviolet-visible (UV-Vis) spectrophotometry TEM, SEM, EDX and FT-IR analyses. Fourier transform infrared analysis revealed the efficient capping and stabilisation properties of these particles and the nature of the capping agent. Silver nanoparticles prepared from leaf extracts showed effective antibacterial activity against Escherichia coli.Conclusion: Biosynthesised AgNPs showed a broad-spectrum antimicrobial susceptibility range and therefore represent promising antimicrobial agents. This is the first reported study for hemp leaf extracts and contributes to the environmentally friendly and cost-effective technique of biosynthesising nanoparticles for drug development.Contribution: This study contributes to the current medicinal properties of cannabis. Furthermore, it reflects that cannabis can also be utilized at a nanoscale with effective antibacterial efficacy.

Antimicrobial and Cytotoxic Effects of Cannabinoids: An Updated Review with Future Perspectives and Current Challenges.

The development of new antibiotics is urgently needed to combat the threat of bacterial resistance. New classes of compounds that have novel properties are urgently needed for the development of effective antimicrobial agents. The extract of Cannabis sativa L. has been used to treat multiple ailments since ancient times. Its bioactivity is largely attributed to the cannabinoids found in its plant. Researchers are currently searching for new anti-infective agents that can treat various infections. Although its phytocannabinoid ingredients have a wide range of medical benefits beyond the treatment of infections, they are primarily associated to psychotropic effects. Different cannabinoids have been demonstrated to be helpful against harmful bacteria, including Gram-positive bacteria. Moreover, combination therapy involving the use of different antibiotics has shown synergism and broad-spectrum activity. The purpose of this review is to gather current data on the actions of Cannabis sativa (C. sativa) extracts and its primary constituents such as terpenes and cannabinoids towards pathogens in order to determine their antimicrobial properties and cytotoxic effects together with current challenges and future perspectives in biomedical application.

Lipid nano-carriers loaded with Cannabis sativa extract for epilepsy treatment – in vitro characterization and in vivo efficacy studies

Taking into consideration the latest reported beneficial anticolvusant effects of cannabidiol (CBD) and cannabiodiolic acid (CBDA) for clinical applications and the advantages of lipid nano-systems as carriers for targeted brain delivery, the aim of this study was set in direction of in vitro physico-chemical and biopharmaceutical characterization and in vivo evaluation of nanoliposomes and nanostructured lipid carriers loaded with Cannabis sativa extract intended for safe and efficient transport via blood-brain barrier and treatment of epilepsy. These nanoliposomes and nanostructured lipid formulations were characterized with z-average diameter <200 nm, following unimodal particle size distribution, negative values for Z-potential, high drug encapsulation efficiency and prolonged release during 24h (38.84-60.91 %). Prepared formulations showed statistically significant higher antioxidant capacity compared to the extract. The results from in vivo studies of the anticonvulsant activity demonstrated that all formulations significantly elevated the latencies for myoclonic, clonic and tonic seizures and, therefore, could be used in preventing different types of seizures. A distinction in the potential of the nano-systems was noted, which was probably anticipated by the type and the characteristics of the prepared formulations.

Are Cannabis and Cannabinoids Effective for Symptomatic Treatment in People With Multiple Sclerosis?: A Cochrane Review Summary With Commentary.

Are Cannabis and Cannabinoids Effective for Symptomatic Treatment in People With Multiple Sclerosis?: A Cochrane Review Summary With Commentary.

Extracellular vesicles of cannabis with high CBD content induce anticancer signaling in human hepatocellular carcinoma.

Plant-derived extracellular vesicles (EVs) have been the topic of interest in recent years due to their proven therapeutic properties. Intact or manipulated plant EVs have shown antioxidant, anti-inflammatory, and anti-cancerous activities as a result of containing bioactive metabolites and other endogenous molecules. Less is known about the EV efficacy with high levels of bioactive secondary metabolites derived from medicinal or non-edible plants. Numerous data suggest the functionality of Cannabis sativa extract and its phytocannabinoids in cancer treatment. Here, two chemotypes of cannabis with different levels of D-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) were selected. EVs were isolated from each chemotype via differential ultracentrifugation. HPLC analysis was illustrative of the absence of THC in EVs derived from both plants. Therefore, two types of EVs were classified according to their CBD content into high- (H.C-EVs) and low-CBD EVs (L.C-EVs). Electron microscopy and DLS showed both cannabis-derived EVs (CDEVs) can be considered as exosome-like nanovesicles. Cytotoxicity assay showed that H.C-EVs strongly decreased the viability of two hepatocellular carcinoma (HCC) cell lines, HepG2 and Huh-7, in a dose and time-dependent manner compared with L.C-EVs. H.C-EVs had no significant effect on HUVECs normal cell growth. The finding showed that the H.C-EVs arrested the G0/G1 phase in the cell cycle and significantly induced cell death by activating mitochondrial-dependent apoptosis signaling pathways in both HCC cell lines. Altogether, the current study highlights that CDEVs can be an ideal natural vehicle for bioactive phytocannabinoids and a promising strategy in cancer management.

Abstract 3714: The antitumor activity of cannabis sativa and CBD in prostate cancer PC3 cells

Abstract Prostate cancer is the second most frequently occurring carcinoma in males worldwide and one of the leading causes of death in men around the world. Recent studies estimate that over 1.4 million males are diagnosed with prostate cancer on an annual basis, with approximately 375 000 succumbing to the disease annually. With current treatments continuing to show severe side effects, there is a need for new treatments. In this study we looked at the effect of cannabis sativa extract, cannabidiol and cisplatin on prostate cancer cells, PC3. Methods: In addressing the above questions, we employed the MTT assay to measure the antiproliferative effect on PC3 cells following treatment with varying concentrations of Cannabis sativa extract, cisplatin and cannabidiol. xCELLigence was also used to confirm the IC50 activity in which cells were grown in a 16 well plate coated with gold and monitor cell. Caspase 3/7 activity was also measured using 96 well-plate following treatment. Western-blot and qRT-PCR was also used to measure the gene expression of tumor suppressor genes, p53, Bax and Bcl2. Animal studies were employed to measure the growth of PC3-mouse derived cancer to evaluate the effect of compounds in vivo. Results: From the treatment with varying concentrations of Cannabis sativa extract, cannabidiol and cisplatin, we have observed that the three compounds induced antiproliferation of PC3 cancer cell lines through the activation of caspase 3/7 activity. We also observed induction of apoptosis in these cells following silencing of retinoblastoma binding protein 6 (RBBP6), with upregulation of p53 and bax mRNA expression, and a reduction in Bcl2 gene expression. The growth of tumors in the mouse models were reduced following treatment with cisplatin and cannabidiol. Conclusion: We demonstrated that cannabidiol is a viable therapy to treat prostate cancer cells, in combination with silencing of RBBP6. This suggests that cannabidiol rather Cannabis sativa extract may play an important role in reducing cancer progression. Citation Format: Lesetja Raymond Motadi, Nonhlanhla B. Moleya. The antitumor activity of cannabis sativa and CBD in prostate cancer PC3 cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3714.

Cannabis sativa exacerbate testicular function by increased oxidative stress, altered male reproductive hormones, sperm quality/quantity and cellular architecture of the testis.

The illicit use of Cannabis in Nigeria is on the increase within the last 10 years and has been implicated as one of the causes of fertility disorder among young men in Nigeria. Hence, this study aimed to ascertain the reproductive impact of n-hexane extract of Cannabis sativa on male Wistar rats. Fifteen male Wistar rats weighing between 180 and 200 g were assigned into three groups of five animals each. The first group served as control, second (low dose), and third (high dose) were fed with n-hexane extract of C. Sativa at doses of 0.75 mg/100 g and 1.5 mg/100 g body weight respectively for 28 days. The administration was done via orogastric. The results revealed the mean serum gonadotropin-releasing hormone (GnRH) concentration in LD and HD was significantly lower (p < 0.05) than in control. LH was significantly higher (p < 0.01) in both LD and HD compared to control. There was a significant increase (p < 0.01) in LD and HD compared to control. Testosterone was significantly elevated in HD (p < 0.05) compared to control. Testicular reduced glutathione and Superoxide dismutase were significantly (p < 0.05) reduced in fed rats compared to control. Testicular catalase and Malondialdehyde were significantly (p < 0.05) increased in HD compared to control. Non-motile sperm count showed a significant (p < 0.001) increase in fed rats compared to control. Total sperm count was significantly (p < 0.05) reduced in fed rats than in control. There were significant (p < 0.001) increased spermatozoa with abnormal morphology in treated groups compared to control. HDL cholesterol was significantly (p < 0.05) elevated in HD compared to control. The atherogenic index was significantly (p < 0.05) reduced in the group fed with 1.5 mg/kg compared to the control. Furthermore, total cholesterol, triglyceride, VLDL-c, and LDL-c did not reveal any significant difference statistically. Histological examination of testes and epididymis of fed rats showed an empty cavity with a reduction in spermatogenesis. This study suggests that n-hexane extract of C. Sativa had more effects on the gonads compared to the higher centres and its ingestion in whatever form could impair male reproductive function.

The modulatory effects of alkaloid extracts of Cannabis sativa, Datura stramonium, Nicotiana tabacum and male Carica papaya on neurotransmitter, neurotrophic and neuroinflammatory systems linked to anxiety and depression.

This study investigated the modulatory effects of alkaloid extracts of Cannabis sativa (CSAE), Datura stramonium (DSAE), Nicotiana tabacum (NTAE) and male Carica papaya (CMAE) on neurotransmitter, neurotrophic and neuro-inflammatory systems linked to anxiety and depression. Male Wistar rats were orally administered the alkaloid extracts in doses of 5, 50, 500, and 2000mg/kg for 90days. On day 91, neurobehavioural studies were evaluated, rats were sacrificed, brain hippocampus removed and tissue homogenate prepared. Biochemical, cytokine and neurotransmitter metabolisms were estimated in the hippocampus. Expressions of genes linked to anxiety and depression were evaluated by RT-qPCR. Results showed CSAE, NTAE and CMAE act as anxiolytic and antidepressant agents by depleting TNF-α, IL-1β and reactive oxygen species concentrations, and monoamine oxidase, angiotensin 1-converting enzyme and acetylcholinesterase activities while elevating IL-10 and dopamine concentrations and glutamate dehydrogenase activity at doses of 5, 50 and 500. Same doses of CSAE, NTAE and CMAE also depleted the gene expressions of GSK3β, JNK, NF-ĸB, and Nesfatin-1 while increasing expressions of CREB, BDNF, serotonin and Nrf2. However, administration of DSAE and 2000mg/kg CSAE, NTAE and CMAE had adverse modulatory effects on the neurochemical concentrations and activities as well as the gene expressions of the evaluated neurotransmitter, neurotrophic and inflammatory systems. In conclusion, the study established the sub-chronic instrumentalization potential of CSAE, CMAE, and NTAE for anxiolytic and anti-depressive moods, though their use may be associated with dependence and addiction, which may result in more detrimental effects than any therapeutic potential they may proffer.

NMR Spectroscopy Applied to the Metabolic Analysis of Natural Extracts of Cannabis sativa.

Cannabis sativa is a herbaceous multiple-use species commonly employed to produce fiber, oil, and medicine. It is now becoming popular for the high nutritional properties of its seed oil and for the pharmacological activity of its cannabinoid fraction in inflorescences. The present study aims to apply nuclear magnetic resonance (NMR) spectroscopy to provide useful qualitative and quantitative information on the chemical composition of seed and flower Cannabis extracts obtained by ultra-sound-assisted extraction, and to evaluate NMR as an alternative to the official procedure for the quantification of cannabinoids. The estimation of the optimal ω-6/ω-3 ratio from the 1H NMR spectrum for the seed extracts of the Futura 75 variety and the quantitative results from the 1H and 13C NMR spectra for the inflorescence extracts of the Tiborszallasi and Kompolti varieties demonstrate that NMR technology represents a good alternative to classical chromatography, supplying sufficiently precise, sensitive, rapid, and informative data without any sample pre-treatment. In addition, different extraction procedures were tested and evaluated to compare the elaboration of spectral data with the principal component analysis (PCA) statistical method and the quantitative NMR results: the extracts obtained with higher polarity solvents (acetone or ethanol) were poor in psychotropic agents (THC < LOD) but had an appreciable percentage of both cannabinoids and triacylgliceroles (TAGs). These bioactive-rich extracts could be used in the food and pharmaceutical industries, opening new pathways for the production of functional foods and supplements.

BACTERIOLOGICAL CHARACTERISTICS OF DIABETIC FOOT INFECTION AND SUSCEPTIBILITY OF MULTIRESISTANT ISOLATES TO HYDRAULIC EXTRACTS FROM ALLIUM SATIVA, ALLIUM CEPA AND CANNABIS SATIVA

Background and objective : Diabetic foot infection is one of the most serious complications of diabetes and its persistence is the result of the ineffectiveness of antibiotic therapy due to the exponentially increasing of antibiotic resistant bacteria. The study aimed at investigating the antibacterial effect of the aqueous extract of some plants on the antibiotic resistant bacteria isolated from diabetic foot wound infections. Methods : A 6-months cross-sectional study from July 2021 to January 2022 at the Yaoundé Central Hospital, was undertaken with diabetic foot wound patients. All samples were appraised to determine presence of infectious agents using standard methods for isolation and identification of bacteria. Subsequently, Antibiotic resistance was done using Kirby-Bauer disc diffusion methods. Finally, extracts of Cannabis sativa leaves, Allium cepa and Allium sativum bulbs were obtained with water and their antibacterial activities were evaluated by the microdillution method on liquid medium. Results : 20 patients whom 14 men were included, with a sex ratio of 2.33, and their mean age was 52.5±9.6 years. 60% of these patients presented wounds in grade III and were of several types : purulent (48.57%), moist (31.43%) and dry (20%). 35 strains were isolated. The predominant GPB were S. aureus (34.29%) followed by Coagulase-negative Staphylococcus (14.29%), and Bacillus Spp (2.86%). Among the GNB, Pseudomonas aeruginosa (11.46%), Serratia Spp. (8.56%), Escherichia coli (8.56%), Enterobacter Spp. (5.71%), Proteus Spp. (5.71%), Klebsiella Spp., Yersinia Spp. and Salmonella Spp. in proportions of 2.86% each. A high rate of antibiotic resistance was recorded for Oxacillin (100%), Vancomycin (83.34%) and Augmentin (55.56%). Sensitivity tests on liquid medium showed that MIC ranged between 3.12-25.00 mg/mL, 6.25-25.00 mg/mL and 1.86-25.00 mg/mL respectivelly for A. sativa, A. cepa and C. sativa. Alliums were much more active on GNB. Although these results are low, they could be an alternative for the diabetic foot infection treatment. Conclusion : Alliums were much more active on GNB Although these results are low, they could be an alternative for the diabetic foot infection treatment. Peer Review History: Received: 28 February 2022; Revised: 15 April; Accepted: 24 April, Available online: 15 May 2022 Academic Editor: Dr. Sally A. El-Zahaby, Pharos University in Alexandria, Egypt, sally.elzahaby@yahoo.com UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 5.5/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Dr. Sangeetha Arullappan, Universiti Tunku Abdul Rahman, Malaysia, sangeetha@utar.edu.my Prof. Dr. Ali Gamal Ahmed Al-kaf, Sana'a university, Yemen, alialkaf21@gmail.com Similar Articles: THE SCOPING REVIEW OF CHINESE AND WESTERN MEDICINE TREATMENT OF DIABETIC FOOT IN ASIA A SURVEY ABOUT THE CURRENT SCENARIO OF DIABETES IN CHATTOGRAM AREA BY DETERMINING PRACTICE OF THE SELF-CARE ACTIVITIES AMONGST PEOPLE WITH DIABETES

Cannabis and cannabinoids for symptomatic treatment for people with multiple sclerosis.

Cannabis and cannabinoids for symptomatic treatment for people with multiple sclerosis.

New AKT-dependent mechanisms of anti-COVID-19 action of high-CBD Cannabis sativa extracts

COVID-19 is caused by the SARS-CoV-2 virus, which enters target cells via interactions with ACE2 and TMPRSS2. Here, we show AKT serine/threonine kinase-dependent epigenetic control of ACE2 and TMPRSS2 expression by high-cannabidiol (CBD) cannabis extracts and their individual components. CBD alone and extracts #1, #5, #7, and #129 downregulated ACE2 and TMPRSS2 in lung fibroblast WI-38 cells through AKT-mediated inhibition. miR-200c-3p and let-7a-5p were two contributing miRNAs in CBD-mediated suppression of ACE2 and TMPRSS2. CBD and terpene PTWT2.2 profoundly inhibited ACE2 and TMPRSS2 expression, both individually and in combination. Extracts #1, #5, #7, and #169 suppressed COX2 expression and remarkably attenuated TNFα/IFNγ-triggered induction of proinflammatory factors IL-6 and IL-8 by AKT pathway. The most abundant molecules present in extracts #1 and #7 modulated the expression of COX2, IL-6, and IL-8 both individually and in combination. These results reveal that high-CBD cannabis extracts attenuated ACE2 and TMPRSS2 expression and the induction of inflammatory mediators COX2, IL-6, and IL-8 via the AKT pathway, highlighting their potential anti-COVID-19 features.

Cannabis with breast cancer treatment: propitious or pernicious?

The online version contains supplementary material available at 10.1007/s13205-021-03102-1.

Non-psychoactive cannabinoids identification by linear retention index approach applied to a hand-portable capillary liquid chromatography platform.

The aim of the present research was the application of the linear retention index (LRI) system for the identification of non-psychoactive cannabinoids using a portable LC instrument. The miniaturization, viz. the use of very low quantities of mobile phase, enabled the development of a compact mobile system to be used for in situ analysis, also according to a green and cost-saving approach. In particular, new capillary LC (cap-LC) methods coupled with UV detection were developed for the analysis of extracts of Cannabis sativa L. Two setups were explored to achieve the efficient separation of twenty-four cannabinoids: a single column setup which exploited a sub-2µm packing to increase the chromatographic resolution, and a dual-column setup based on the serial connection of two different stationary phases, each coupled to an UV detector. The latter allowed the determination of two LRI values for each analyte, thus increasing the identification power. Moreover, since two different wavelengths were used on the LED-based UV detectors, the ratio of the absorbances measured on each chromatographic trace represented a third identification criterion, thus fulfilling the recommendations of the Scientific Working Group for The Analysis of Seized Drugs (SWDRUG) about the categories of analytical techniques to be used and the minimum number of parameters required for the unambiguous identification of drugs. The obtained results could be used for the development of a novel analytical method for fast and automatic in situ forensic investigations and hemp breeding programs, also minimizing the consumption of both sample and solvent.

In vitro effects of cannabidiol and its main metabolites in mouse and human Sertoli cells

Cannabidiol (CBD) is a major cannabinoid present in extracts of the plant Cannabis sativa (marijuana). While the therapeutic effects of CBD on epilepsy have been demonstrated, less is understood regarding its potential adverse effects. Recent studies revealed that CBD induced toxicity in the male reproductive system of animal models. In this study, we used TM4, an immortalized mouse Sertoli cell line, and primary human Sertoli cells to evaluate the toxicities of CBD and its main metabolites, 7-carboxy-CBD and 7-hydroxy-CBD. CBD induced concentration- and time-dependent cytotoxicity in mouse and human Sertoli cells, which mainly resulted from the inhibition of the G1/S-phase cell cycle transition. CBD also inhibited DNA synthesis and downregulated key cell cycle proteins. Moreover, CBD reduced the mRNA and protein levels of a functional marker, Wilms’ tumor 1. Similar to CBD, 7-carboxy-CBD and 7-hydroxy-CBD inhibited cellular proliferation and decreased DNA synthesis. 7-Carboxy-CBD was less cytotoxic than CBD, while 7-hydroxy-CBD showed comparable cytotoxicity to CBD in both mouse and human Sertoli cells. Compared to mouse Sertoli cells, CBD, 7-hydroxy-CBD, and 7-carboxy-CBD were more cytotoxic in human Sertoli cells. Our results indicate that CBD and its main metabolites can inhibit cell proliferation in mouse and human Sertoli cells.

In Vivo Evaluation of Cannabis sativa Full Extract on Zebrafish Larvae Development, Locomotion Behavior and Gene Expression.

Historically, humans have been using Cannabis sativa for both recreational and medical purposes. Nowadays, cannabis-based products have gained scientific interest due to their beneficial effects on several syndromes and illnesses. The biological activity of cannabinoids is essentially due to the interaction with the endocannabinoid system, and zebrafish (Danio rerio) is a very well-known and powerful in vivo model for studying such specific interactions. The aim of the study was to investigate the effects of different doses of a Cannabis sativa whole extract [dissolved in dimethyl sulfoxide (DMSO)] on zebrafish eggs’ hatchability, embryo post-hatching survival, larvae locomotion behavior and mRNA gene expression. The results showed the absence of toxicity, and no significant differences were observed between treatments for both embryo hatching and survival rate. In addition, larvae exposed to the cannabis extract at the highest dose [containing 1.73 nM and 22.3 nM of ∆9-tetrahydrocannabinol (THC) and cannabidiol (CBD), respectively] showed an increased locomotion compared to the control and DMSO treated groups. Moreover, qRT-PCR analysis showed that the highest dosage of cannabis induced an over-expression of cnr1 and cnr2 cannabinoid receptors. In conclusion, the exposition of zebrafish larvae to the whole extract of Cannabis sativa showed no negative effects on embryo development and survival and enhanced the larvae’s locomotor performances. These findings may open up possible Cannabis sativa applications in human pharmacology as well as in other animal sectors.

The use of medical cannabis in pediatric palliative care: a case series

BackgroundMedical cannabis may be a useful tool for managing treatment-resistant epilepsy and chronic pain, which affect many patients in pediatric palliative care (PPC); however, little evidence is available in this setting.Case presentationWe aimed to describe a clinical experience in a setting where high-level evidence may not be obtained. We report our clinical experience in a pediatric palliative care department in Italy. Caregivers reported changes in intensity and frequency of pain and epilepsy events. Six patients received a titrated plant extract of cannabis sativa for 1 year. Only mild and transient adverse events occurred: drowsiness, euphoria, restlessness and tachycardia; the resolution was either spontaneous or obtained by modifying the administration schedule. Treatment was never discontinued. No overdoses occurred. All patients experienced seizures during the pre-treatment observation period, and obtained a reduction in seizure frequency, although with variable extent while receiving cannabis. In addition, a benefit on pain was observed, based on the caregiver’s evaluation, and a reduction of analgesic use.ConclusionOur experience suggests that a titrated plant extract preparation of medical cannabis may be useful to control treatment-resistant pain and epilepsy in PPC patients.