Efectos de la marihuana en la salud bucal / Effects of marijuana on oral health

Aun cuando las autoridades del Sector Salud en México no se han declarado respecto al uso medicinal de la marihuana, con el objetivo de conocer el estado actual internacional sobre sus riesgos y usos terapéuticos, investigamos los avances reportados en la actualidad, así como las comunidades que han despenalizado su uso. Se presenta su origen como elemento terapéutico, pueblos involucrados, diversas denominaciones, historicidad, las diversas preparaciones, farmacodinamia, sus efectos nocivos a la salud en general y particularmente en boca, sus posibles usos en odontología tomando en cuenta sus propiedades terapéuticas. ampliamente reseñadas en relación a otros lugares del organismo. Finalmente, la propuesta de investigación en odontología con especial énfasis en aquellas especialidades donde la inflamación y el dolor agudo estén presentes de manera significativa (AU)

Although health authorities in Mexico have not officially declared their stance on the medicinal use of marijuana, our research aims to explore the current international status regarding its risks and therapeutic uses. We have investigated the latest reported advancements and examined communities that have decriminalized its usage. This presentation encompasses its therapeutic origin, involved communities, various designations, historical context, diverse preparations, pharmacodynamics, its adverse effects on overall health and particularly oral health, as well as its potential applications in dentistry, considering its widely documented therapeutic properties in comparison to other areas of the body. Finally, our research proposal in dentistry places special emphasis on specialties where inflammation and acute pain are significantly present (AU)

Structural Basis for Molecular Recognition of Cannabinoids by Inhibitory Cys-Loop Channels.

Cannabis sativa has a long history of medicinal use, dating back to ancient times. This plant produces cannabinoids, which are now known to interact with several human proteins, including Cys-loop receptors for glycine (GlyR) and gamma-aminobutyric acid (GABAAR). As these channels are the primary mediators of inhibitory signals, they contribute to the diverse effects of cannabinoids on the nervous system. Evidence suggests that cannabinoid binding sites are located within the transmembrane domain, although their precise location has remained undetermined for over a decade. The process of identification of the binding site and the computational approaches employed are the main subjects of this Perspective, which includes an analysis of the most recently resolved cryo-EM structures of zebrafish GlyR bound to Δ9-tetrahydrocannabinol and the THC-GlyR complex obtained through molecular dynamics simulations. With this work, we aim to contribute to guiding future studies investigating the molecular basis of cannabinoid action on inhibitory channels.

Cytogenotoxicity and inflammatory response in liver of rats exposed to different doses of cannabis nano emulsions.

Cannabis is the most used illicit substance for recreational purposes around the world. However, it has become increasingly common to witness the use of approved cannabis preparations for symptoms management in various diseases. The aim of this study was to investigate the effects of cannabis nano emulsion in the liver of Wistar rats, with different proportions of delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD). For this, a total of 40 male Wistar rats were distributed into 5 groups, as follows (n = 8 per group): Control: G1, Experimental group (G2): treated with cannabis nano emulsion (THC and CBD) at a dose of 2.5 mg/kg, Experimental group (G3): treated with cannabis nano emulsion (THC and CBD) at a dose of 5 mg/kg, Experimental group (G4): treated with cannabis nano emulsion (CBD) at a dose of 2.5 mg/kg; Experimental group (G5): treated with cannabis nano emulsion (CBD) at a dose of 5 mg/kg. Exposure to the nano emulsion was carried out for 21 days, once a day, orally (gavage). Our results showed that cannabis nano emulsions at higher doses (5 mg/kg), regardless of the composition, induced histopathologic changes in the liver (G3 and G5) in comparison with the control group. In line with that, placental glutathione S-transferase (GST-P) positive foci increased in both G3 and G5 (p < 0.05), as well as the immune expression of Ki-67, vascular endothelial growth factor (VEGF) and p53 (p < 0.05). Also, the nano emulsion intake induced an increase in the number of micronucleated hepatocytes in G5 (p < 0.05) whereas G3 showed an increase in binucleated cells (p < 0.05). As for metanuclear alterations, karyolysis and pyknosis had an increased frequency in G3 (p < 0.05). Taken together, the results show that intake of cannabis nano emulsion may induce degenerative changes and genotoxicity in the liver in higher doses, demonstrating a clear dose-response relationship.

Determination of Cannabinoids in Cannabis sativa Oil and Infused Ice Cream by LC-DAD Method.

BACKGROUND: Cannabis sativa is known to produce a class of terpenophenolic compounds named cannabinoids. The two main ones are cannabidiol (CBD) and tetrahydrocannabinol (THC), which have therapeutic properties. In the development of cannabis-based preparations, it is important to have suitable analytical methods for the analysis of the principal cannabinoids. OBJECTIVE: This study aimed to develop and validate a simple and rapid HPLC method with photodiode array detection for determination of CBD and THC in Cannabis sativa oil extract and infused ice cream, including a stability study. METHOD: Chromatographic separation of CBD and THC was performed with a C18 column, with a mobile phase consisting of acetonitrile and water with formic acid (80 + 20 v/v) in isocratic elution mode, with detection at 208 nm for CBD and 280 nm for THC and 1.0 mL/min flow rate. RESULTS: The method was linear over a range of 1-5 µg/mL for CBD, and 20-100 µg/mL for THC; the relative standard deviation was <3.6%, the recovery ranged between 98.8 and 102.5% for oil and between 84 and 94% for ice cream, QL was 0.33 µg/mL for CBD and 2.30 µg/mL for THC, and the assay demonstrated adequate selectivity. CBD and THC were stable for at least 28 days under light protection at 22°C, 4°C, and -20°C in the oil and for at least 60 days at -20°C in the ice cream. CONCLUSIONS: The results showed that the method was suitable for quantitative determination of CBD and THC in Cannabis sativa oil extract and infused ice cream, and it is useful for quality control purposes. HIGHLIGHTS: The method is simple and fast, and it is useful for the quality control of a new product corresponding to an ice cream based on a Cannabis sativa oil extract.

E-cigarette use-associated lung injury (EVALI).

The prevalence of vaping has overtaken conventional cigarettes as the most frequent form of nicotine consumption among 15-24-year olds. There are currently a large number of both legitimate and illegitimate products and suppliers offering more than 8000 different flavors of vape on the market, whose additives are not tested, studied or regulated and whose safety and toxicity profile remains unknown. In vitro studies have demonstrated a dose-dependent decrease in the viability of normal human bronchial epithelial cells after exposure to vapor from electronic vape devices.Short- and medium-term studies to date indicate that vapor-induced pulmonary lesions are the most serious and commonly reported side effect; such lesions include bilateral ground glass opacities in lung bases with subpleural preservation, bilateral infiltrates, pleural effusion, pneumomediastinum and nodular opacities. Cases of EVALI have been described in patients with daily exposure, as well as in users who reported having been exposed to these substances at least once a month. The most frequently inhaled substances are THC, flavored liquids of unknown content, and nicotine.The clinical manifestations of dyspnea and cough are the most frequent respiratory symptomatology, in addition to constitutional manifestations such as fever and chills, and gastrointestinal manifestations such as vomiting, nausea, abdominal pain and diarrhea. To these can be added the presence of tachypnea, tachycardia, elevated blood pressure, hypoxia, leukocytosis with neutrophilia and elevated ESR.

Potency trends of cannabis in Jamaica during the period of 2014 to 2020.

Reports suggest that cannabis potency has dramatically increased over the last decade in the USA and Europe. Cannabinoids are the terpeno-phenolic compounds found in the cannabis plant and are responsible for its pharmacological activity. The two most prominent cannabinoids are delta-9-tetrahydrocannabinol (Δ9 THC) and cannabidiol (CBD). Cannabis potency is measured not only by the Δ9 THC levels but also by the ratio of Δ9 THC to other non-psychoactive cannabinoids, namely, CBD. Cannabis use was decriminalized in Jamaica in 2015, which opened the gates for the creation of a regulated medical cannabis industry in the country. To date, there is no information available on the potency of cannabis in Jamaica. In this study, the cannabinoid content of Jamaican-grown cannabis was examined over the period 2014-2020. Two hundred ninety-nine herbal cannabis samples were received from 12 parishes across the island, and the levels of the major cannabinoids were determined using gas chromatography-mass spectrometry. There was a significant increase (p < 0.05) in the median total THC levels of cannabis samples tested between 2014 (1.1%) and 2020 (10.2%). The highest median THC was detected in the central parish of Manchester (21.1%). During the period, THC/CBD ratios increased from 2.1 (2014) to 194.1 (2020), and there was a corresponding increase in the percent freshness of samples (CBN/THC ratios <0.013). The data show that a significant increase in the potency of locally grown cannabis has occurred in Jamaica during the last decade.

Efficacy and safety of medical cannabinoids in children with cerebral palsy: a systematic review.

INTRODUCTION: The increasing popularity of cannabinoids for treating numerous neurological disorders has been reported in various countries. Although it reduces tetrahydrocannabinol psychoactivity, it helps patients tolerate higher doses and complements the anti-spasmodic effects of tetrahydrocannabinol. One of the most important potential of cannabinoids are related to its potential to help children with cerebral palsy, a contributor of lifelong disability. Therefore, this systematic review aimed to assess the efficacy and safety of medical cannabinoids in children with cerebral palsy. METHODS: This review adhered to The Preferred Reporting Items for Systematic Reviews and Meta-analysis 2020 guidelines. Seven databases, namely, Scopus, PubMed, EBSCO Host, ProQuest, Google Scholar, Semantic Scholar, and JSTOR, were used to identify relevant studies. Studies examining pediatric patients with cerebral palsy and reporting the efficacy and safety of medical cannabinoids through clinical trials, observational cross-sectional studies, or cohort designs were included. The outcomes of the studies included the efficacy of medical cannabinoids administered for spasticity, motor components, pain control, sleep difficulties, adverse effects, and seizure control. RESULTS: Of 803 identified articles, only three met the inclusion criteria for data synthesis. One study exhibited a moderate risk-of-bias. A total of 133 respondents, mainly from Europe, were investigated. Overall effectiveness and safety were considered good. However, the results are inconsistent, especially regarding spasticity treatment variables. CONCLUSION: The anti-spasticity, anti-inflammatory, and anti-seizure properties of cannabinoids might be beneficial for patients with cerebral palsy, although their effectiveness has not been widely studied. Further studies with larger sample sizes and various ethnicities are warranted. Prospero database registration: (www.crd.york.ac.uk/prospero) under ID CRD42022358383.

Beneficio clínico del tratamiento con cannabinoides para el dolor crónico no oncológico / Clinical benefit of cannabinoids to treat chronic non-cancer pain / Benefício clínico do tratamento com canabinoide para dor crônica não oncológica

Introducción: los cannabinoides pueden ser una opción válida para el tratamiento del dolor crónico no oncológico de acuerdo a los estudios publicados hasta el momento y a nuestra experiencia clínica. Objetivo: valorar el beneficio clínico de preparados de cannabis medicinal (CM) para dolor crónico no oncológico en pacientes que consultaron en la Clínica de Endocannabinología del Uruguay (CEDU). Material y método: estudio descriptivo, observacional, longitudinal, de una población atendida en un centro privado de salud. Se trata de una cohorte de 438 pacientes que consultaron espontáneamente en CEDU desde septiembre de 2016 a marzo de 2020. El motivo de consulta fue dolor crónico no oncológico que no respondió al tratamiento estándar. Resultados: en la cohorte estudiada predominaron las mujeres (74%), promedio 69 años, que se asisten en el sistema privado de salud en el 95% de los casos, en su mayoría con instrucción secundaria. El tipo de dolor más frecuente fue el dolor osteoarticular. El quimiotipo de CM más usado fue cannabidiol (CBD) al 5%, con buena respuesta al tratamiento en el descenso del nivel del dolor y suspensión o disminución de uso de opioides (y derivados) y antiinflamatorios no esteroideos (AINES). Se observaron escasos y leves efectos adversos (EA) en la gran mayoría de los pacientes. Abandonaron el tratamiento 12 pacientes (menos del 3%). Conclusiones: esta investigación retrospectiva mostró una caída del nivel del dolor de 3,14 (valor p ≤ 0,0001), indicando que el CM puede ser una opción para el tratamiento del dolor crónico no oncológico. Se requieren más estudios para demostrar la efectividad y seguridad de los cannabinoides. Esto depende de muchos factores (leyes que faciliten la accesibilidad a variedad de productos de CM de grado médico, incentivos a la ciencia e investigación). De todas formas, podemos afirmar que los resultados presentados son prometedores en relación con su potencial terapéutico.

Introduction: Cannabinoids can be a valid option for the treatment of chronic non-cancer pain, according to the studies published to date and our clinical experience. Objectives: To evaluate the clinical benefit of medicinal cannabis preparations (MCPs) for chronic non-cancer pain in patients seen at the Endocannabinology Clinic of Uruguay (CEDU). Method: Descriptive, observational, longitudinal study of a population treated at a private healthcare center. This involves a cohort of 438 patients who spontaneously consulted at CEDU from September 2016 to March 2020. The reason for consultation was chronic non-cancer pain that did not respond to standard treatment. Results: in the studied cohort, women prevailed and accounted for 74% of patients. Average age was 69 years old and 95% of them sought care within the private healthcare system. Most women had completed secondary school education. The most frequent type of pain was osteoarticular pain. The most used chemovar of Medicinal Cannabis (MC) was 5% cannabidiol (CBD), showing a favorable treatment response in reducing pain levels and the discontinuation or reduction of opioid and non-steroidal anti-inflammatory drug (NSAID) usage. Few and mild adverse effects (AE) were observed in the vast majority of patients. Twelve patients (less than 3%) discontinued the treatment. Conclusions: This retrospective study demonstrated a reduction in pain level of 3.14 (p-value ≤ 0.0001) indicating that MC could be an option for the treatment of non-oncological chronic pain. Further studies are needed to demonstrate the effectiveness and safety of cannabinoids. This depends on many factors (laws facilitating accessibility to a variety of medical-grade MC products, incentives for science and research). Nevertheless, we can assert that the presented results are promising in consideration of their therapeutic potential.

Introdução: os canabinoides podem ser uma opção válida para o tratamento da dor crônica não oncológica de acordo com estudos publicados até o momento e nossa experiência clínica. Objetivos: avaliar o benefício clínico das preparações de Cannabis Medicinal (CM) para dor crônica não oncológica em pacientes que consultaram a Clínica de Endocanabinologia do Uruguai (CEDU). Método: estudo descritivo, observacional, longitudinal de uma população atendida em um centro de saúde privado. Esta é uma coorte de 438 pacientes que consultaram espontaneamente no CEDU no período setembro de 2016 - março de 2020. O motivo da consulta foi dor crônica não oncológica que não respondeu ao tratamento padrão. Resultados: na coorte estudada, 74% eram mulheres, a idade média foi 69 anos, 95% frequentam a rede privada de saúde e a maioria com ensino médio. O tipo de dor mais frequente foi a osteoarticular. O quimiotipo de MC mais utilizado foi o Canabidiol 5% (CBD), com boa resposta ao tratamento em termos de redução do nível de dor e suspensão ou redução do uso de opioides (e derivados) e anti-inflamatórios não esteroides (AINEs). A grande maioria dos pacientes apresentou poucos e leves efeitos adversos (EAs). Menos de 3% dos 12 pacientes abandonou o tratamento. Conclusões: Esta investigação retrospectiva mostrou uma queda no nível de dor de 3,14 (valor de p ≤ 0,0001), indicando que o MC pode ser uma opção para o tratamento da dor crônica não oncológica. São necessários mais estudos para demonstrar a eficácia e segurança dos canabinoides. Isso depende de muitos fatores (leis que facilitem o acesso a uma variedade de produtos CM de grau médico, incentivos para ciência e pesquisa). De qualquer forma, podemos afirmar que os resultados apresentados são promissores em relação ao seu potencial terapêutico.

Development and Validation of a Simple, Fast, and Accessible HPLC-UV Method for Cannabinoids Determination in Cannabis sativa L. Extracts and Medicinal Oils.

INTRODUCTION: Cannabis sativa L. is a well-recognized medicinal plant. Cannabis regulations in Argentina are insufficient to solve the problem of patient access to full-spectrum cannabis-based products. So, the market of artisanal products with unknown quality and dosage of cannabinoids is increasing, and so is the local demand and need for analyzing these products. However, much of the latest validated methodologies for cannabinoid quantification include expensive instrumentation that is not always available in laboratories of health institutions in Argentina. METHODS: The aim of this work was to develop and validate a simple and rapid HPLC-UV method for the identification and quantification of principal cannabinoids in cannabis resins, inflorescences, and medicinal oils using standard HPLC equipment. The cannabinoids selected for validation were cannabidiol acid (CBDA), cannabigerol (CBG), cannabidiol (CBD), cannabinol (CBN), delta-9-tetrahydrocannabinol (Δ9-THC), cannabichromene (CBC), and tetrahydrocannabinol acid (THCA). A method for the simultaneous identification and quantification of these 7 main cannabinoids was developed and then validated. Some data parameters were comparable to other reports with more sophisticated analytical instruments for the analysis of cannabis. The assessed limits of detection and the limits of quantitation ranged from 0.9 to 3.66 µg/mL and 2.78 to 11.09 µg/mL, respectively. The concentration-response relationship of the method indicated a linear relationship between the concentration and peak area with R2 values of > 0.99 for all 7 cannabinoids. RESULTS: The relative standard deviation (RSD%) varied from 2.34 to 4.82 for intraday repeatability and from 1.16 to 3.15 for interday repeatability. The percentage of recovery values was between 94 to 115% (resins) and 80 to 103% (inflorescence extract). The cannabis industry is growing rapidly, and there is a need for reliable testing methods to ensure the safety and efficacy of cannabis products. In addition, current methods for cannabinoid analysis are often time-consuming and expensive, while the HPLC-UV method herein reported is a simple, rapid, accurate, and cost-effective alternative for the analysis of cannabinoids in cannabis resins, inflorescences, and medicinal oils. CONCLUSION: This method will be proposed to be included in the Cannabis sativa L. monograph of the Argentine Pharmacopoeia.

An assessment of qualitative and quantitative cannabinoids analysis in selected commercially available cannabis oils in Argentina.

In recent years, the therapeutic use of cannabis products, especially cannabis oils, has increased significantly, due to the pharmacological potential of their cannabinoids, for the treatment of conditions, such as pain management, cancer, and epilepsy. In Argentina, patients with medical prescriptions can access to cannabis oil, through self-cultivation, a third-person (grower or importer), or a civil organization authorized for that purpose. However, these products remain largely unregulated in Argentina, and information available regarding labeling accuracy, especially cannabidiol (CBD)/ Δ9-tetrahydrocannabinol (Δ9-THC) concentrations are inconsistent or nonexistent, nor long-term product stability, and lot to lot variability. Understanding these properties is fundamental if these products are to be used in patients with a determinate pathology. Therefore, we analyzed commercially available cannabis oils (n: 500) in Argentina for qualitative and quantitative cannabinoids content. In order to provide a detailed overview of their cannabinoids profiles, and determine Δ9-THC, CBD, and cannabinol (CBN) concentrations, samples were diluted and analyzed by gas chromatography- mass spectrometry (GC/MS). Most of the samples tested positive for cannabinoids (n: 469) with Δ9-THC and CBD as the predominant cannabinoids. Among products tested, only 29.8% (n: 149) gave specific CBD label claims, and testing indicated a CBD tested positive of 70.5% (n: 105). For products (n: 17) with a THC-free label claim, testing indicated 76.5% (n: 13) of Δ9-THC positive, and cannabinoids were not detected in four products. Δ9-THC concentrations ranged from 0.1 to 143.0 mg/mL, CBD concentrations from 0.1 to 125.3 mg/mL, and CBN concentrations from 0.04 to 60.10 mg/mL; CBN/ Δ9-THC ratios ranged from 0.0012 to 2.31, and CBD/ Δ9-THC ratios from 0.0008 to 178.87. Furthermore, the (Δ9-THC + CBN)/CBD ratio of most samples was greater than one. In summary, our results indicate that cannabis oil products show wide variability in cannabinoids content, purity, and labeling.

Cannabis: Drug of Abuse and Therapeutic Agent, Two Sides of the Same Coin.

The consumption of Cannabis sativa plant, known as marijuana in the Western world, for different purposes (therapeutic, intoxicating, and spiritual) due to its psychoactive effects, can be traced back to ancient times. Cannabis is the most used illicit drug worldwide; however, its legal status is changing rapidly. Cannabis regulation will allow a better understanding of its effects as a misused drug, including new challenges, such as the availability of highly potent Cannabis extracts. Furthermore, scientific research is making significant efforts to take advantage of the potential therapeutic uses of Cannabis active compounds. The science of Cannabis derivatives started with the identification of the phytocannabinoids Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD), allowing the formal study of the complex set of effects triggered by Cannabis consumption and the deciphering of its pharmacology. Δ9-THC is recognized as the compound responsible for the psychoactive and intoxicating effects of Cannabis. Its study led to the discovery of the endocannabinoid system, a neuromodulatory system widespread in the human body. CBD does not induce intoxication and for that reason, it is the focus of the search for cannabinoid potential clinical applications. This review examines the current state of knowledge about contrasting perspectives on the effects of Cannabis, Δ9-THC, and CBD: their abuse liability and potential therapeutic use; two sides of the same coin.

Determination of amphetamines, opioids, cocaine metabolites and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in urine using biocompatible solid-phase microextraction and ultra-performance LC-MS/MS.

Background: Workplace drug testing primarily relies on urine analysis, targeting multiple compounds with varying physicochemical characteristics. Biocompatible solid-phase microextraction (BioSPME) is a miniaturized solid-phase extraction technique that enables the simultaneous extraction and preconcentration of analytes directly from the biological matrix. Methods: The BioSPME procedure consisted of the sequential extraction of 50-µl urine samples using LC Tips C18 in basic and acidic pH, followed by desorption with methanol and n-hexane, respectively. The extracts were analyzed by ultra-performance LC-MS/MS. Results: Intra-day precision was 1.2-8.6% and inter-day precision was 1.8-14.2%. Accuracy was 96.8-107.4%. The extraction yields were 62.8-109.4%. The matrix effects were -3.98% to 1%. Conclusion: BioSPME shows promise as an alternative method for preparing urine samples prior to drug measurement by ultra-performance LC-MS/MS.

Cannabis sativa-based oils against aluminum-induced neurotoxicity.

The use of terpenoid compounds in different neural-related conditions is becoming useful for several illnesses. Another possible activity of these compounds is the reduction of nervous impairment. Cannabis sativa plants are known for their concentration of two important terpenoids, the delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD). CBD and THC have central peripheral activities already described and their usage in different brain diseases, such as Alzheimer's and multiple sclerosis. Aluminum (Al) is known as an important neurotoxic compound, the physiological action of Al is not known already, and in high concentrations can lead to intoxication and cause neurotoxicity. Here we evaluated the potential effect of two different doses of CBD- and THC-rich based oils against Al-induced toxicity, in the zebrafish model. We evaluated behavioral biomarkers of the novel tank test (NTT) and social preference test (SPT), and biochemical markers: the activity of the enzyme acetylcholinesterase (AChE) and the antioxidant enzymes-catalase, superoxide dismutase, and glutathione-S-transferase. CBD- and THC-based oils were able to increase the AChE activity helping the cholinergic nervous system actuate against Al toxicity which was reflected by the behavioral biomarkers changes. We concluded that the oils have a protective effect and might be used with proposals for neurological and antioxidant impairment avoidance caused by Al intoxications.

Argentination: A Silver Bullet for Cannabinoid Separation by Differential Mobility Spectrometry.

As the legality of cannabis continues to evolve globally, there is a growing demand for methods that can accurately quantitate cannabinoids found in commercial products. However, the isobaric nature of many cannabinoids, along with variations in extraction methods and product formulations, makes cannabinoid quantitation by mass spectrometry (MS) challenging. Here, we demonstrate that differential mobility spectrometry (DMS) and tandem-MS can distinguish a set of seven cannabinoids, five of which are isobaric: Δ9-tetrahydrocannabinol (Δ9-THC), Δ8-THC, exo-THC, cannabidiol, cannabichromene, cannabinol, and cannabigerol. Analytes were detected as argentinated species ([M + Ag]+), which, when subjected to collision-induced dissociation, led to the unexpected discovery that argentination promotes distinct fragmentation patterns for each cannabinoid. The unique fragment ions formed were rationalized by discerning fragmentation mechanisms that follow each cannabinoid's MS3 behavior. The differing fragmentation behaviors between species suggest that argentination can distinguish cannabinoids by tandem-MS, although not quantitatively as some cannabinoids produce small amounts of a fragment ion that is isobaric with the major fragment generated by another cannabinoid. By adding DMS to the tandem-MS workflow, it becomes possible to resolve each cannabinoid in a pure N2 environment by deconvoluting the contribution of each cannabinoid to a specific fragmentation channel. To this end, we used DMS in conjunction with a multiple reaction monitoring workflow to assess cannabinoid levels in two cannabis extracts. Our methodology exhibited excellent accuracy, limits of detection (10-20 ppb depending on the cannabinoid), and linearity during quantitation by standard addition (R2 > 0.99).

A case report of CBD and THC as analgesic therapy in a cat with chronic osteoarthritic pain.

A 10-year-old mixed breed male cat presented with clinical signs related to chronic orthopaedic pain. Upon physical examination, pain was noted, based on the feline Musculoskeletal Pain Index (FMPI). An analgesic treatment with a full spectrum cannabis oil (1.8% CBD and 0.8% THC) was proposed for 30 days (0,5 mg/kg based on CBD). The FMPI scale score decreased more than 50%. This case reported a satisfactory outcome for the patient and the owner, although this medication could increase ALT. Given the paucity of literature published to date on the treatment of veterinary species with cannabis-based medications, further clinical and pharmacokinetic studies are necessary to study the safety and efficacy of its use.

Effects of delta-9 tetrahydrocannabinol on fear memory labilization and reconsolidation: A putative role of GluN2B-NMDA receptor within the dorsal hippocampus.

Cannabis preparations could be an effective reconsolidation-based treatment for post-traumatic stress disorder. However, the effects of Δ9-tetrahydrocannabinol (THC) in fear memory labilization, a critical condition for retrieval-induced reconsolidation, are undetermined. We sought to investigate the effect of a conventional and an ultra-low dose of THC in memory labilization of adult male Wistar rats submitted to contextual fear conditioning. Pretreatment with THC 0.002, but not THC 0.3 mg/kg, i. p., before memory retrieval, did not change memory expression during the retrieval but impaired reconsolidation. No treatment changed freezing expression in an unpaired context. Before retrieval, THC 0.3, but not THC 0.002, decreased GluN2A-NMDA expression and the GluN2A/GluN2B ratio in the dorsal hippocampus (DH) 24 h later. No changes were observed immediately after retrieval. Pretreatment with THC 0.3 abolished the reconsolidation-impairing effect of anisomycin injected into the DH, suggesting an impairment in memory labilization. This effect was associated with an increased freezing expression in the unpaired context and was not observed with the THC ultra-low dose. The GluN2B-NMDA antagonism increased fear generalization in the anisomycin-treated group but restored its reconsolidation-impairing effect and reduced fear generalization when animals were pretreated with THC 0.3. GluN2A-NMDA antagonism or inhibition of the ubiquitin-proteasome system in the DH did not interfere with the effects of THC 0.3. Our findings indicate that THC causes a bidirectional effect on fear memory labilization that depends on hippocampal GluN2B-NMDA receptors' involvement in fear memory generalization.

Molecular study of endo and phytocannabinoids on lipid membranes of different composition.

The discovery of the endocannabinoid system (ECS) dates back only 30 years. Although many research groups have been elucidating its components, location, functions and metabolism, the peculiarities of the compounds considered "neurotransmitters" of ECS generate questions that have not yet been answered or controversies in the literature. In this context, we studied the molecular behaviour of the main endocannabinoid compounds and the main phytocannabinoids in eukaryotic outer and inner model membranes. The high lipophilicity of these compounds gives place to the hypothesis that cannabinoids may reach the molecular targets through the lipid bilayer. This consideration is not only for the cannabinoid receptors but also for other (many) targets that these bioactive molecules modulate (Watkins, 2019; Nelson et al., 2020; Jakowiecki and Filipek, 2016). Given the reported multitarget action of these compounds and the differential behaviour towards the different receptors, studying the properties and dynamics of these cannabinoids in POPC and POPE model membranes become relevant. In this regard, we have studied the differential modulation of the endocannabinoids anandamide and 2-arachidonoyl-glycerol and the phytocannabinoids cannabidiol and trans-Δ9-tetrahydrocannabinol to eukaryotic outer and inner model membranes. Results show that behaviours favour the mobility of the bioactive molecules studied by the external eukaryotic model membrane. As well as, the internal eukaryotic model membrane is less fluid, favouring the stabilisation of folded conformations or the positioning of the molecules in the centre of the bilayer. These results provide relevant evidence that contributes to a deep inside understanding of the behaviour of the primary endogenous ligands of ECS, together with the principal phytocannabinoids of C. sativa.

Entourage Effect and Analytical Chemistry: Chromatography as a Tool in the Analysis of the Secondary Metabolism of Cannabis sativa L.

Cannabis sativa L. has been used as medicine for thousands of years. Since the early identification of tetrahydrocannabinol (THC) in 1960, pharmacological activities were attributed to a group of unique structures named cannabinoids. For decades, research and development were applied to determine different cannabinoids and their medicinal properties. Nowadays there is evidence that the therapeutic benefits of the plant are based on the synergy of cannabinoids and other secondary metabolites such as terpenes and flavonoids. Differences between the medical performance of isolated compounds like cannabidiol (CBD) or THC and full-spectrum plant extracts are notable. Indeed, the superiority of the last one is provoked by the synergy between various different compounds. This improved medicinal effect is called the entourage effect. Chromatography has become the method of choice for the determination of cannabinoids, terpenes, and flavonoids, so it represents an excellent tool for a proper characterization of the plant and plant derived products. The objective of characterization relies not only in analyzing the fingerprint of cannabis, but also to identify different chemotypes for medical purposes. To understand the contributions of each natural product to this "entourage effect", this review presents an in-depth analysis of the utilization of High-performance liquid chromatography (HPLC), Gas chromatography (GC) and other methods for the analysis of phytocomponents of Cannabis sativa L. In this sense, a representative number of examples and advances made in the field together with limitations and future needs are provided. It can be concluded that standardized protocols and quality control policies and procedures are necessary for the comprehensive analysis of cannabis extracts and derivatives.

Molecular determinants of tetrahydrocannabinol binding to the glycine receptor.

The recognition of Cannabis as a source of new compounds suitable for medical use has attracted strong interest from the scientific community in its research, and substantial progress has accumulated regarding cannabinoids' activity; however, a thorough description of their molecular mechanisms of action remains a task to complete. Highlighting their complex pharmacology, the list of cannabinoids' interactors has vastly expanded beyond the canonical cannabinoid receptors. Among those, we have focused our study on the glycine receptor (GlyR), an ion channel involved in the modulation of nervous system responses, including, to our interest, sensitivity to peripheral pain. Here, we report the use of computational methods to investigate possible binding modes between the GlyR and Δ9 -tetrahydrocannabinol (THC). After obtaining a first pose for the THC binding from a biased molecular docking simulation and subsequently evaluating it by molecular dynamic simulations, we found a dynamic system with an identifiable representative binding mode characterized by the specific interaction with two transmembrane residues (Phe293 and Ser296). Complementarily, we assessed the role of membrane cholesterol in this interaction and positively established its relevance for THC binding to GlyR. Lastly, the use of restrained molecular dynamics simulations allowed us to refine the description of the binding mode and of the cholesterol effect. Altogether, our findings contribute to the current knowledge about the GlyR-THC mode of binding and propose a new starting point for future research on how cannabinoids in general, and THC in particular, modulate pain perception in view of its possible clinical applications.

Effects of Δ9-THC and Type-1 Cannabinoid Receptor Agonists in the Elevated Plus Maze Test of Anxiety: A Systematic Review and Meta-Analysis.

Δ9-THC (the main active compound from Cannabis sativa) and related cannabinoids have been used as drugs of abuse and as medications. They induce a complex set of emotional responses in humans and experimental animals, consisting of either anxiolysis or heightened anxiety. These discrepant effects pose a major challenge for data reproducibility and for developing new cannabinoid-based medicines. In this study, we review and analyze previous data on cannabinoids and anxiety-like behavior in experimental animals. Systematic review and meta-analysis on the effects of type-1 cannabinoid receptor agonists (full or partial, selective or not) in rodents exposed to the elevated plus maze, a widely used test of anxiety-like behavior. Cannabinoids tend to reduce anxiety-like behavior if administered at low doses. THC effects are moderated by the dose factor, with anxiolytic- and anxiogenic-like effects occurring at low-dose (0.075-1 mg/kg) and high-dose (1-10 mg/kg) ranges, respectively. However, some studies report no effect at all regardless of the dose tested. Finally, motor impairment represents a potential confounding factor when high doses are administered. The present analysis may contribute to elucidate the experimental factors underlying cannabinoid effects on anxiety-like behavior and facilitate data reproducibility in future studies.