Cannabis Sativa Research Articles

Fertiliser supplementation with a PGR complex of chlormequat chloride (CCC) and paclobutrazol (PBZ) alters Cannabis sativa growth

ABSTRACT Cannabis (Cannabis sativa) is a high-value crop, wherein growth within controlled environments is common for the purposes of maximising yield, quality, and uniformity, however, uncontrolled-growth in proximity to artificial lighting can compromise yields. Within the floricultural industry it is common practice to apply plant growth regulators (PGRs) to retard plant growth and maximise yield and quality, with two well-established PGRs being chlormequat chloride (CCC) and paclobutrazol (PBZ). Accordingly, the aim of this study was to investigate the potential of a PGR complex of CCC and PBZ for novel application to cannabis to control plant height. Treatment of cannabis with the PGR complex within an indoor hydroponic growth system resulted in a 91.1% reduction in post-treatment plant growth (p < 0.001) leading to plants with end-point height and internodal spacing (INS) 0.568-fold (p < 0.001) and 0.469-fold (p < 0.001) shorter than control plants, respectively. These results demonstrate the capacity of CCC and PBZ to beneficially halt the growth of cannabis. Accordingly, application of these PGRs in combination may be utilised to restrict cannabis height without compromising yield within commercial cultivation.

Direct Inhibition of BK Channels by Cannabidiol, One of the Principal Therapeutic Cannabinoids Derived from Cannabis sativa.

Cannabidiol (CBD), one of the main Cannabis sativa bioactive compounds, is utilized in the treatment of major epileptic syndromes. Its efficacy can be attributed to a multimodal mechanism of action that includes, as potential targets, several types of ion channels. In the brain, CBD reduces the firing frequency in rat hippocampal neurons, partly prolonging the duration of action potentials, suggesting a potential blockade of voltage-operated K+ channels. We postulate that this effect might involve the inhibition of the large-conductance voltage- and Ca2+-operated K+ channel (BK channel), which plays a role in the neuronal action potential's repolarization. Thus, we assessed the impact of CBD on the BK channel activity, heterologously expressed in HEK293 cells. Our findings, using the patch-clamp technique, revealed that CBD inhibits BK channel currents in a concentration-dependent manner with an IC50 of 280 nM. The inhibition is through a direct interaction, reducing both the unitary conductance and voltage-dependent activation of the channel. Additionally, the cannabinoid significantly delays channel activation kinetics, indicating stabilization of the closed state. These effects could explain the changes induced by CBD in action potential shape and duration, and they may contribute to the observed anticonvulsant activity of this cannabinoid.

In silico investigation of cannabinoids from Cannabis sativa leaves as a potential anticancer drug to inhibit MAPK-ERK signaling pathway and EMT induction

Genes related to MAPK-ERK signaling pathways, and epithelial-mesenchymal transition induction is evolutionarily conserved and has crucial roles in the regulation of important cellular processes, including cell proliferation. In this study, six cannabinoids from Cannabis sativa were docked with MAPK-ERK signaling pathways to identify their possible binding interactions. The results showed that all the cannabinoids have good binding affinities with the target proteins. The best binding affinities were MEK- tetrahydrocannabinol (– 8.8 kcal/mol) and P13k-cannabinol (– 8.5 kcal/mol). The root mean square deviation was calculated and used two alternative variants (rmsd/ub and rmsd/lb) and the values of rmsd/lb fluctuated 8.6–2.0 Å and for rmsd/ub from 1.0 to 2.0 Å that suggests the cannabinoids and protein complex are accurate and cannot destroy on binding. The study analyzed the pharmacokinetic and drug-likeness properties of six cannabinoids from C. sativa leaves using the SwissADME web tool. Lipinski's rule of five was used to predict drug-likeness and showed that all compounds have not violated it and the total polar surface area of cannabinoids was also according to Lipinski's rule that is benchmarked of anticancer drugs. Cannabinoids are meet the requirements of leadlikeness and synthetic accessibility values showed they can be synthesized. The molecular weight, XLOGP3, solubility (log S), and flexibility (FLEX) are according to the bioavailability radar. The bioavailability score and consensus Log Po/w fall within the acceptable range for the suitable drug. Pharmacokinetics parameters showed that cannabinoids cannot cross the blood–brain barrier, have high GI absorption as well as cannabinoids are substrates of (CYP1A2, CYP2C19, CYP2C9, CYP2D6, and CYP3A4) but no substrate of P-glycoprotein. Based on these findings, the study suggests that cannabinoids are suitable drugs that could be used as effective inhibitors for target proteins involved in cancer pathways. Among the six cannabinoids, cannabinol and tetrahydrocannabinol exerted maximum binding affinities with proteins of MAPK-ERK signaling pathways, and their pharmacokinetics and drug-likeness-related profiles suggest that these cannabinoids could be superlative inhibitors in cancer treatment. Further in vitro, in vivo, and clinical studies are needed to explore their potential in cancer treatment.

Aqueous Extracts of Cannabis sativa: Cytotoxicity and Effects on Angiogenesis, Oxidative Stress and Nitric Oxide Production

AbstractCannabis sativa has been used in local medicine to manage cancer, ache, inflammation, diabetes, and other conditions. The aqueous extract of Cannabis sativa leaves collected from two geographical locations (South Africa, EC and Lesotho, LS) was assessed for their potential against angiogenesis, nitric oxide (NO) production, oxidative stress and cytotoxicity. The EC extract showed better angiogenesis and vascular endothelial growth factor suppression than the LS extract at 20 and 100 µg/mL. EC and LS displayed their highest NO inhibitory effects (91 and 76%) at the most negligible 1.6 µg/mL concentration. The glutathione and Catalase levels increased significantly in the MCF-7 cells following administration with EC and LS. In contrast, a decrease in the malondialdehyde (MDA) level in the cells' supernatant was found compared to the untreated cells. High-performance liquid chromatography analysis reveals the presence of cannabidiol (CBD) and tetrahydrocannabinoilic acid (THCA), with EC and LS having a considerable number of similar peaks. The extracts were relatively non-cytotoxic within the concentration range evaluated (0.78125 -100–µg/mL), though selective effects on cell survival at some concentrations were observed. The study provides information to support the extensive folkloric use of the aqueous extract of C. sativa in cancer and other associated ailments, as well as diseases linked to oxidative stress. Studies on the aqueous extract using animal models or other relevant in vivo models are recommended.

The Comparison of Physical and Chemical Properties of Pellets and Briquettes from Hemp (Cannabis sativa L.)

The Comparison of Physical and Chemical Properties of Pellets and Briquettes from Hemp (Cannabis sativa L.)

Terpenes from Cannabis sativa induce antinociception in a mouse model of chronic neuropathic pain via activation of adenosine A2A receptors.

Terpenes are small hydrocarbon compounds that impart aroma and taste to many plants, including Cannabis sativa. A number of studies have shown that terpenes can produce pain relief in various pain states in both humans and animals. However, these studies were methodologically limited and few established mechanisms of action. In our previous work, we showed that the terpenes geraniol, linalool, β-pinene, α-humulene, and β-caryophyllene produced cannabimimetic behavioral effects via multiple receptor targets. We thus expanded this work to explore the potential antinociception and mechanism of these Cannabis terpenes in a mouse model of chronic pain. We first tested for antinociception by injecting terpenes (200 mg/kg, IP) into male and female CD-1 mice with mouse models of chemotherapy-induced peripheral neuropathy (CIPN) or lipopolysaccharide-induced inflammatory pain, finding that the terpenes produced roughly equal antinociception to 10 mg/kg morphine or 3.2 mg/kg WIN55,212. We further found that none of the terpenes produced reward as measured by conditioned place preference, while low doses of terpene (100 mg/kg) combined with morphine (3.2 mg/kg) produced enhanced antinociception vs either alone. We then used the adenosine A2A receptor (A2AR) selective antagonist istradefylline (3.2 mg/kg, IP) and spinal cord-specific CRISPR knockdown of the A2AR to identify this receptor as the mechanism for terpene antinociception in CIPN. In vitro cAMP and binding studies and in silico modeling studies further suggested that the terpenes act as A2AR agonists. Together these studies identify Cannabis terpenes as potential therapeutics for chronic neuropathic pain and identify a receptor mechanism for this activity.

Hemp Seeds (Cannabis sativa L.) as a Valuable Source of Natural Ingredients for Functional Foods—A Review

Hemp Seeds (Cannabis sativa L.) as a Valuable Source of Natural Ingredients for Functional Foods—A Review

Extraction, purification and in vitro assessment of the antioxidant and anti-inflammatory activity of policosanols from non-psychoactive Cannabis sativa L

Policosanols (PCs) are bioactive compounds extracted from different natural waxes. In this work, the purification, characterization and assessment of the antioxidant and anti-inflammatory activity was carried out on PCs from an innovative source, i.e. a waxy material from supercritical-fluid extraction (SFE) of non-psychoactive Cannabis sativa L. (hemp) inflorescences. Starting from this material, PCs were obtained by microwave-assisted trans-esterification and hydrolysis, followed by preparative liquid chromatography under normal phase conditions. The purified product was characterized using high-performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). In vitro cell-free and cell-based antioxidant and anti-inflammatory assays were then performed to assess their bioactivity.HPLC-ELSED analysis of the purified mixture from hemp wax revealed C26OH and C28OH as the main compounds. In vitro assays indicated an inhibition of intracellular reactive oxygen species (ROS) production, a reduction of nuclear factor kappa B (NF-κB) activation and of the activity of the neutrophil elastase. Immunoblotting assays allowed us to hypothesize the mechanism of action of the compounds of interest, given the higher levels of MAPK-activated protein kinase 2 (MK2) and heme oxygenase-1 (HO-1) protein expression in the PCs pretreated HaCaT cells. In conclusion, even if more research is needed to unveil other molecular mechanisms involved in hemp PCs activity, our results suggest that these compounds may have potential for use in oxinflammation processes.

Dual-step pharmacological intervention for traumatic-like memories: implications from D-cycloserine and cannabidiol or clonidine in male and female rats.

Therapeutic approaches to mitigating traumatic memories have often faced resistance. Exploring safe reconsolidation blockers, drugs capable of reducing the emotional valence of the memory upon brief retrieval and reactivation, emerges as a promising pharmacological strategy. Towards this objective, preclinical investigations should focus on aversive memories resulting in maladaptive outcomes and consider sex-related differences to enhance their translatability. After selecting a relatively high training magnitude leading to the formation of a more intense and generalized fear memory in adult female and male rats, we investigated whether two clinically approved drugs disrupting its reconsolidation remain effective. We found resistant reconsolidation impairment by the α2-adrenergic receptor agonist clonidine or cannabidiol, a major non-psychotomimetic Cannabis sativa component. However, pre-retrieval administration of D-cycloserine, a partial agonist at the glycine-binding site of the N-methyl-D-aspartate (NMDA) receptor complex, facilitated their impairing effects on reconsolidation. A similar reconsolidation blockade by clonidine or cannabidiol was achieved following exposure to a non-conditioned but generalized context after D-cycloserine administration. This suggests that sufficient memory destabilization can accompany generalized fear expression. Combining clonidine with cannabidiol without potentiating memory destabilization by D-cycloserine was ineffective. These findings highlight the importance of NMDA receptor signaling in memory destabilization and underscore the efficacy of a dual-step pharmacological intervention in attenuating traumatic-like memories, even in a context different from the original learning environment.

Analysis of the Physicochemical Properties of Hemp Seed Oil Extracted from Roasted Hemp Seeds (Cannabis sativa L.)

Analysis of the Physicochemical Properties of Hemp Seed Oil Extracted from Roasted Hemp Seeds (<i>Cannabis sativa</i> L.)

Potential Anti-Tumorigenic Properties of Diverse Medicinal Plants against the Majority of Common Types of Cancer

Potential Anti-Tumorigenic Properties of Diverse Medicinal Plants against the Majority of Common Types of Cancer

Exploring the potentialities of industrial hemp for sustainable rural development

This literature review paper explores the potential sustainable applications of industrial hemp, contextualizing it within the UN Agenda 2030 Sustainable Development Goals (SDGs). Hemp is a dioecious, herbaceous, anemophilous (wind-pollinated) annual crop that grows from one to five meters in height. However, monoecious varieties have been developed through breeding and selection. Historically versatile, Cannabis sativa L. has faced stigmatization and unfounded prohibitions, having played crucial roles in different civilizations, including during World War I. A member of the Cannabaceae family, it is a versatile non-psychoactive crop, with low concentrations of THC. Recently, the global market for hemp has grown. It is an eco-friendly option in the food, textile, pharmaceutical, and energy sectors. This study is based on bibliographic research covering primary and secondary sources, including technical reports, academic journals, and scientific repositories. The origin, characteristics and cultivation methods of industrial hemp are discussed, besides its practical applications, including the production of fibers, health food grains, medicinal phytocannabinoids, biofuels, sustainable biomass energy, and carbon offsetting. Its potential to contribute to sustainable agriculture and achieve SDG targets stands out, considering environmental benefits, product diversification, resistance, and integration into agroforestry systems. Regenerative agriculture is explored as a promising approach, aligned with the principles of sustainable agriculture and the SDGs, aiming to restore agricultural ecosystems, promote soil health, and improve climate resilience. The research highlights the importance of reforming cannabis-related policies to achieve goals relating to health, justice, and global partnerships, emphasizing the need for sustainable approaches to exploiting this natural resource.

Usability of Hemp Plant in Poultry Nutrition

Hemp (Cannabis sativa L.) is an annual plant with 2n=20 chromosomes, one-year, C3 group, cultivated for its long and strong fibers and seeds. It is a plant that should be cultivated in a controlled manner due to the presence of THC in its natural structure. Producers who avoid control and monitoring have turned to alternative products instead of hemp farming. Countries that cultivate cannabis in large areas; France, China, and Canada. Some countries are updating their cannabis-related laws and making efforts to increase hemp acreage. It is known that hemp farming has been practiced in Anatolia since 1500 BC. Various products such as hemp seeds, hemp pulp, hemp oil have been used in various studies in poultry feeding studies, while no negative results have been encountered, but various positive results have been suggested and their use is recommended. It is of great importance to investigate the effects of cannabidiols, which have strong antimicrobial, antioxidant and immunostimulant effects, as they are expected to support the immune system in poultry, especially broilers. Some of the studies in which hemp products are used in poultry nutrition are as follows; Addition of hemp meal to laying hen diets has been reported to make no significant difference in egg production, feed consumption, feed utilization, body weight gain or egg quality, but results in lower palmitic acid concentrations and higher Linoleic acid (LA) and alpha-linolenic acid (ALA) concentrations, which are healthier for human consumption. Hemp seeds and hemp oil used in chicken rations caused an increase in the omega-3 polyunsaturated fatty acid content and color intensity of egg yolks, but no adverse effects were observed on the sensory profiles of cooked eggs. Addition of cannabis meal to fast growing broiler diets did not affect performance or mortality, and no effect on the number of Clostridium perfringens in the cecum was reported. Hemp seeds have a remarkable effect on the growth of broiler chicks and can help reduce feed expenditures for broiler rearing. In this paper, the use and usability of the hemp plant, whose production areas are expanding rapidly, in poultry will be examined.

Draft genome sequences for four isolates of the hemp (Cannabis sativa) fungal pathogen Neofusicoccum parvum.

Four isolates of Neofusicoccum parvum, collected from diseased hemp (Cannabis sativa) plants over a period of 2 years and shown to be pathogenic on C. sativa, were examined in this study. Their genome sizes ranged between 42.8 and 44.4 Mb, with 16,499 ± 72 predicted genes across the four isolates.

The artificial production of viral-like particles in Nicotiana benthamiana suggests the pro-assembly role of the Cannabis cryptic virus RdRP

AbstractThe partitivirids feature an icosahedral protein coating accommodating both their dsRNA genome and RNA-dependent RNA polymerase (RdRP). This signifies that transcription and replication activities of the viral polymerase occur within the capsid, emphasizing that the viral cycle relies on polymerase incorporation. Particles lacking RdRP are defective and hence non-infectious. Encapsidation and replication are intricately linked for dsRNA viruses, to the extent that, for many of these, such as the cystovirids, the RdRP serves a dual role as a transcriptase/replicase and a pro-assembly factor, ensuring structural stability and overall capsid integrity. This work investigates if RdRP has a similar role within the capsid of Cannabis cryptic virus (CanCV), a betapartitivirus affecting Cannabis sativa. Utilizing reverse genetics in Nicotiana benthamiana, we conclusively established that RdRP expression is indispensable for CanCV’s virus-like particle formation. This study enhances our understanding of CanCV encapsidation, with RdRP serving a pivotal role as a pro-assembly factor. These preliminary findings contribute to the knowledge of viral assembly within the Partitiviridae family.

Cannabis sativa Boosts Brain Functions and Ameliorate Anxiety, when used Sensibly and not Abusively

Cannabis sativa is an illegal substance with documented analgesic, antipyretic, anti-inflammatory, and anti-diarrheal properties. Numerous other cannabinoids found in cannabis have an impact on brain activity. So, the purpose of this study was to examine how cannabis sativa extract affected the cognito-motor activity in rats. Twenty-four (24) Wistar rats weighing between 80.53 and 100.49g were randomly divided into four groups (A, B, C, and D) of six animals each after a 14-day acclimatization period to laboratory conditions. Only food and water were provided to Group A. Groups B and C received an oral ethanolic extract of Cannabis sativa at doses of 400 mg/kg and 800 mg/kg, respectively. Group D received an intraperitoneal dose of diazepam at a rate of 50 mg/kg. To ascertain the impact of the cannabis sativa extract on the neuro behavior of the rats, each animal in the four groups is subjected to an experimental test following each administration of the extract (the Barnes maze test, the beam walk test, and the inverted screen tests). The rats in groups B and C that received cannabis sativa extract showed a notable improvement in learning and memory, and the majority of the rats in group C who received cannabis sativa extract walked upright without stumbling. Group A, the control group, exhibited indicators of anxiety and panic but no discernible change in learning or memory. Due to muscle weakness, members of group D (standard drug group) were essentially inactive throughout the experiment, staggering and falling. It can be concluded that cannabis sativa's ethanolic leaf extract is not harmful, boosts brain functions, and promotes learning and memory when used sensibly and not abusively. Thus, it can be inferred that the plant may help with the treatment of several mental health conditions, such as anxiety.

From dawn 'til dusk: daytime progression regulates primary and secondary metabolism in Cannabis glandular trichomes.

Cannabis sativa L. glandular trichomes (GTs) synthesise large amounts of secondary metabolites, predominantly cannabinoids and terpenoids. The associated demand for carbon and energy makes GTs strong sink tissues with indications that their secondary metabolism is coupled to the availability of photoassimilates. Many metabolites show diurnal patterns of flux, but it is unknown whether cannabinoids and terpenoids are regulated by time of day. We quantified cannabinoids, terpenoids and the GT proteome over a 12-hour light period in flowers of Hindu Kush, a high-tetrahydrocannabinol (THC) cultivar. Major cannabinoids changed significantly over the course of day, resulting in an increase in total measured cannabinoids. Major terpenoids also changed, with sesquiterpenes generally decreasing with day progression. While monoterpenes generally did not decrease, the second most abundant, α-pinene, increased. The GT proteome changed the most within the first six hours of the day and analysis of differentially abundant proteins indicated upregulation of primary metabolism. Surprisingly, key cannabinoid biosynthetic enzymes decreased with daytime progression despite increases in cannabinoid content, which indicate that daytime increases of photoassimilates are the main driver of cannabinoid regulation. This first reporting of variability of cannabinoid and terpenoid biosynthesis over the course of the day has implications for Cannabis research and production.

Genome-wide identification and analysis of ascorbate peroxidase (APX) gene family in hemp (Cannabis sativa L.) under various abiotic stresses.

Ascorbate peroxidase (APX) plays a critical role in molecular mechanisms such as plant development and defense against abiotic stresses. As an important economic crop, hemp (Cannabis sativa L.) is vulnerable to adverse environmental conditions, such as drought, cold, salt, and oxidative stress, which lead to a decline in yield and quality. Although APX genes have been characterized in a variety of plants, members of the APX gene family in hemp have not been completely identified. In this study, we (1) identified eight members of the CsAPX gene family in hemp and mapped their locations on the chromosomes using bioinformatics analysis; (2) examined the physicochemical characteristics of the proteins encoded by these CsAPX gene family members; (3) investigated their intraspecific collinearity, gene structure, conserved domains, conserved motifs, and cis-acting elements; (4) constructed a phylogenetic tree and analyzed interspecific collinearity; and (5) ascertained expression differences in leaf tissue subjected to cold, drought, salt, and oxidative stresses using quantitative real-time-PCR (qRT-PCR). Under all four stresses, CsAPX6, CsAPX7, and CsAPX8 consistently exhibited significant upregulation, whereas CsAPX2 displayed notably higher expression levels under drought stress than under the other stresses. Taken together, the results of this study provide basic genomic information on the expression of the APX gene family and pave the way for studying the role of APX genes in abiotic stress.

Non-destructive assessment of cannabis quality during drying process using hyperspectral imaging and machine learning

Cannabis sativa L. is an industrially valuable plant known for its cannabinoids, such as cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC), renowned for its therapeutic and psychoactive properties. Despite its significance, the cannabis industry has encountered difficulties in guaranteeing consistent product quality throughout the drying process. Hyperspectral imaging (HSI), combined with advanced machine learning technology, has been used to predict phytochemicals that presents a promising solution for maintaining cannabis quality control. We examined the dynamic changes in cannabinoid compositions under diverse drying conditions and developed a non-destructive method to appraise the quality of cannabis flowers using HSI and machine learning. Even when the relative weight and water content remained constant throughout the drying process, drying conditions significantly influenced the levels of CBD, THC, and their precursors. These results emphasize the importance of determining the exact drying endpoint. To develop HSI-based models for predicting cannabis quality indicators, including dryness, precursor conversion of CBD and THC, and CBD : THC ratio, we employed various spectral preprocessing methods and machine learning algorithms, including logistic regression (LR), support vector machine (SVM), k-nearest neighbor (KNN), random forest (RF), and Gaussian naïve Bayes (GNB). The LR model demonstrated the highest accuracy at 94.7–99.7% when used in conjunction with spectral pre-processing techniques such as multiplicative scatter correction (MSC) or Savitzky–Golay filter. We propose that the HSI-based model holds the potential to serve as a valuable tool for monitoring cannabinoid composition and determining optimal drying endpoint. This tool offers the means to achieve uniform cannabis quality and optimize the drying process in the industry.

Evaluating the Metabolomic Profile and Anti-Pathogenic Properties of Cannabis Species

The Cannabis species is one of the potent ancient medicinal plants acclaimed for its medicinal properties and recreational purposes. The plant parts are used and exploited all over the world for several agricultural and industrial applications. For many years Cannabis spp. has proven to present a highly diverse metabolomic profile with a pool of bioactive metabolites used for numerous pharmacological purposes ranging from anti-inflammatory to antimicrobial. Cannabis sativa has since been an extensive subject of investigation, monopolizing the research. Hence, there are fewer studies with a comprehensive understanding of the composition of bioactive metabolites grown in different environmental conditions, especially C. indica and a few other Cannabis strains. These pharmacological properties are mostly attributed to a few phytocannabinoids and some phytochemicals such as terpenoids or essential oils which have been tested for antimicrobial properties. Many other discovered compounds are yet to be tested for antimicrobial properties. These phytochemicals have a series of useful properties including anti-insecticidal, anti-acaricidal, anti-nematicidal, anti-bacterial, anti-fungal, and anti-viral properties. Research studies have reported excellent antibacterial activity against Gram-positive and Gram-negative multidrug-resistant bacteria as well as methicillin-resistant Staphylococcus aureus (MRSA). Although there has been an extensive investigation on the antimicrobial properties of Cannabis, the antimicrobial properties of Cannabis on phytopathogens and aquatic animal pathogens, mostly those affecting fish, remain under-researched. Therefore, the current review intends to investigate the existing body of research on metabolomic profile and anti-microbial properties whilst trying to expand the scope of the properties of the Cannabis plant to benefit the health of other animal species and plant crops, particularly in agriculture.