Synthetic Cannabinoids Research Articles

Towards better prepared prisons: monitoring and response to the use of new substances inside prison

Abstract The use of new psychoactive substances (NPS) became an emerging issue in prisons in a number of European countries in 2014-2015. Synthetic cannabinoids were the new psychoactive substances most often reported, followed by synthetic cathinones, synthetic opioids and new benzodiazepines. Quantitative qualitative data on prison and drugs provided by 27 European Member States, Norway and Turkey to the EMCDDA (European Monitoring Centre for Drugs and Drug Addiction). An exploratory study conducted in 2017 found signs of NPS use in prison in 22 countries. Based on 2021-2022 data from a few countries, the lifetime prevalence of NPS in prison ranges from <1 to 19%, with large variation by country. Anecdotal information from 2024 referred to the use of new strong synthetic opioids (nitazene) inside prison. A wide range of physical and mental health harms (e.g. psychosis, disorientation, suicidal ideation, aggressiveness to others or self-harm) has been associated with acute intoxication by, and chronic consumption of NPS inside prison. Recent overdoses associated with the use of nitazene were reported in some prisons in Ireland and the UK. Additionally, the use of NPS in prison can have a disturbing impact on prison management, internal violence and bullying. To date, interventions to tackle the problems related to the use of NPS in prison have tended to focus on supply reduction and controls. Health and social interventions have begun to emerge, but they remain limited. Despite signs of increasing challenges posed by the emergence of NPS since several years, prisons remain largely unprepared to monitor and respond to those challenges. A new key task of the revised EMCDDA mandate (EUDA - European Drug Agency from 2nd of July) is to be better prepared to monitor, forecast and respond to new challenges in the drug field. Prisons should be prepared to adequately address the new challenges posed by the NPS to benefit the health and security of people living and working in prison.

Hydroponic Cultivation of Spices and Aromatics: Techniques, Benefits and Challenges

The global shift towards urbanization, with 55% of the population residing in urban areas and an anticipated rise to 68% by 2050, presents challenges to traditional farming. The increasing world population, further intensifies the strain on available agricultural land due to urban expansion. Climate change combines with these issues, make conventional cultivation more challenging. Alternative agricultural practices, such as hydroponics, gain significance in addressing these concerns. Aromatic and spices have been used for centuries for their medicinal and culinary properties. They contain various essential oils, flavonoids, and other phytochemicals responsible for their distinctive flavour and aroma, as well as their health benefits. The rising global demand for natural products from herbs and spices has led to its market growth. Hydroponics, a technology for cultivating plants in nutrient solutions with or without an artificial medium such as sand, gravel, vermiculite, rockwool, peat moss, coir or sawdust, offers a promising solution. This paper explores the potential of hydroponic cultivation for planting material production, enhancing the yield, nutritional composition, essential oil content and medicinal properties of various spice and aromatic crops, including black pepper, saffron, ginger, garlic, coriander, mint, celery, fenugreek, parsley, thyme, rosemary, green mustard, basil and others. The main challenges in hydroponic production of these crops include high initial investment, precise monitoring and lack of technical knowledge. Hence by reviewing recent literature, this paper aims to provide insights into the classification of hydroponics and its different techniques, benefits and challenges of hydroponic cultivation, various growing media and nutrient solutions used, types of hydroponics utilized and their purposes in the production of diverse herbs and spices. Embracing hydroponic techniques could contribute to sustainable agricultural practices, particularly in urbanized environments where traditional farming faces constraints.

Cannabinoid hyperemesis syndrome: genetic susceptibility to toxic exposure

Cannabinoid hyperemesis syndrome presents as a complex of symptoms and signs encompassing nausea, vomiting, abdominal pain, and hot water bathing behavior, most typically in a heavy cannabis user. Its presentation is frequently associated with hypothalamic-pituitary-adrenal axis activation with stress and weight loss. Recent investigation has identified five statistically significant mutations in patients distinct from those of frequent cannabis users who lack the symptoms, affecting the TRPV1 receptor, two dopamine genes, the cytochrome P450 2C9 enzyme that metabolizes tetrahydrocannabinol, and the adenosine triphosphate-binding cassette transporter. The syndrome is associated with escalating intake of high potency cannabis, or alternatively, other agonists of the cannabinoid-1 receptor including synthetic cannabinoids. Some patients develop environmental triggers in scents or foods that suggest classical conditioned responses. Various alternative “causes” are addressed and refuted in the text, including exposure to pesticides, neem oil or azadirachtin. Nosological confusion of cannabinoid hyperemesis syndrome has arisen with cyclic vomiting syndrome, whose presentation and pathophysiology are clearly distinct. The possible utilization of non-intoxicating antiemetic cannabis components in cannabis for treatment of cannabinoid hyperemesis syndrome is addressed, along with future research suggestions in relation to its genetic foundation and possible metabolomic signatures.

Cannabinoids-Multifunctional Compounds, Applications and Challenges-Mini Review.

Cannabinoids represent a highly researched group of plant-derived ingredients. The substantial investment of funds from state and commercial sources has facilitated a significant increase in knowledge about these ingredients. Cannabinoids can be classified into three principal categories: plant-derived phytocannabinoids, synthetic cannabinoids and endogenous cannabinoids, along with the enzymes responsible for their synthesis and degradation. All of these compounds interact biologically with type 1 (CB1) and/or type 2 (CB2) cannabinoid receptors. A substantial body of evidence from in vitro and in vivo studies has demonstrated that cannabinoids and inhibitors of endocannabinoid degradation possess anti-inflammatory, antioxidant, antitumour and antifibrotic properties with beneficial effects. This review, which spans the period from 1940 to 2024, offers an overview of the potential therapeutic applications of natural and synthetic cannabinoids. The development of these substances is essential for the global market of do-it-yourself drugs to fully exploit the promising therapeutic properties of cannabinoids.

Assessment of pharmacological effects and abuse potential of 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201 in mice.

The newly emerging synthetic cannabinoids (SCs) 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201 have been observed to produce effects by activating cannabinoid type 1 (CB1) receptors. Nevertheless, the pharmacological effects and potential for abuse of these three substances remain to be studied. These substances have yet to be regulated in many countries. We investigated the safety, pharmacological effects, rewarding effects, and cannabinoid withdrawal of 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201. This study evaluated the drug safety and the cannabinoid-specific pharmacological effects of the three substances through acute toxicity experiments (in which the LD50 of each substance was obtained) and tetrad experiments (comprising assessments of hypothermia, analgesia, locomotion inhibition, and catalepsy). Furthermore, the conditioned place preference (CPP) experiments and withdrawal experiments were conducted to evaluate the rewarding effect and cannabinoid withdrawal potential of the substances in question. The results demonstrated that all three drugs exhibited certain acute toxic effects and could potentially induce tetrad effects. The data were analyzed using non-linear regression, and the corresponding ED50 values and 95% confidence intervals (CI) were obtained. The rank order of potency was determined to be CUMYL-PEGACLONE > 5F-EDMB-PICA > NM-2201. In the CPP experiments, it was demonstrated that 5F-EDMB-PICA significantly induced an increase in CPP score at a dose of 0.3mg/kg, while NM-2201 caused an increase in CPP score and a significant aversion effect at a dose of 2 and 3mg/kg, respectively. It is noteworthy that all three types of SCs were observed to produce a significant biphasic effect, indicating that CPP scores were biphasic for all compounds. Following the administration of the CB1 receptor antagonist rimonabant, a notable increase in head twitches and paw tremors was observed, indicating that these three SCs induce cannabinoid withdrawal through the mediation of CB1 receptors. The results of this study indicated that these SCs possess cannabinoid-specific pharmacological effects and abuse potential, which provides substantial experimental data to support the future regulation of these substances.

Waxy- or Putty-Like Materials as a Novel Drug Preparation for Synthetic Cannabinoid Receptor Agonists: Detection in Prisons and InVitro Cannabinoid Receptor Activity.

After the Scottish Prison Service (SPS) introduced mail photocopying procedures in December 2021, a shift in smuggling methods was observed for synthetic cannabinoid receptor agonists (SCRAs) and other new psychoactive substances (NPS) from drug-infused papers back to traditional sample matrices (e.g., tablets and powders), although new matrices also emerged. This study reports on waxy- or putty-like materials as a novel drug preparation for SCRAs and other drugs seized from UK prisons. In 2023, 22 of these new preparations were seized from Scottish prisons, with eight found in sealed vape pods. The materials were positive for SCRAs, phytocannabinoids, novel benzodiazepines, and/or gabapentinoids. Additionally, 11 preparations were seized from an English prison, all containing the SCRAs MDMB-4en-PINACA and MDMB-INACA. MDMB-INACA was pharmacologically characterized using invitro CB1 and CB2 bioassays, revealing moderate efficacy but low potency at CB1. Furthermore, the invitro CB1 bioassay was also used to evaluate the CB1 activating potential of extracts from eight seized samples. Six of these showed high CB1 activity, whereas the samples lacking SCRAs or containing only MDMB-INACA showed no or only weak CB1 activity, respectively. Lastly, applying the bioassay as an activity-based "untargeted" screening method effectively identified the presence of SCRAs in one waxy preparation, which was initially not detected by gas chromatography-mass spectrometry (GC-MS). This underscores the effectiveness of the bioassay for evaluating these new waxy- or putty-like materials for the presence of SCRAs.

Are the postmortem concentration changes of the synthetic cannabinoid cumyl-5F-P7AICA and its N-pentanoic acid metabolite dependent on the environmental conditions? – A systematic study following pulmonary administration to pigs

The number of fatal cases involving synthetic cannabinoids (SCs) is increasing. However, interpretation of postmortem (PM) toxicological findings is challenging, due to unknown PM intervals and possible redistribution processes or instabilities. Only sparse data on SCs are available. Therefore, a controlled pig study was performed to determine the PM stability of cumyl-5F-P7AICA under different environmental conditions. Ten pigs inhalatively received 200 µg/kg body weight cumyl-5F-P7AICA each. Six hours later, they were euthanized and biopsies of the main tissues and body fluids were taken. Animals were stored in air or water (n=5 each) and samples were repeatedly taken for three days. Quantification of cumyl-5F-P7AICA and its N-pentanoic acid metabolite (NPA) was performed using standard addition or a fully validated method (blood) followed by LC-MS/MS. Time-dependent concentration changes of cumyl-5F-P7AICA were observed in liver, bile fluid and muscle specimens at both conditions. Concentrations of NPA only changed considerably in duodenum content of animals stored in air. Environment-dependent concentrations changes were only observed for cumyl-5F-P7AICA in kidney and the NPA metabolite in duodenum content. Overall, cumyl-5F-P7AICA and its metabolite seem to be quite stable in PM specimens. Hence, also central blood might be analyzed, if no peripheral blood is available.

Ocular effects of synthetic cannabinoids: a case-control study.

To evaluate the ocular effects of "Bonzai", a synthetic cannabinoid (SC), in seropositive and seronegative users. Sixty eyes of 60 consecutive male patients with a history of "Bonzai" use and 30 eyes of 30 age-matched male healthy controls were enrolled in this case-control study. Patients with past "Bonzai" use were grouped as seropositive (n:30) and seronegative (n:30) according to urine toxicology tests. All groups were compared for blood pressures, intraocular pressure, foveal and parafoveal retinal thicknesses, subfoveal and parafoveal choroidal thicknesses, measurements of the peripapillary retinal nerve fibre layer (RNFL), and macular ganglion cell complex (GCC), subfoveal total choroidal, luminal and stromal areas, and choroidal vascularity index (CVI). No differences were noted in blood pressures between the groups (p > 0.05). The mean intraocular pressure was significantly lower in the seropositive group than in the other groups (p < 0.001). Foveal and retinal thicknesses, RNFL, and GCC measurements did not differ between the groups (p > 0.05). Subfoveal and parafoveal choroidal thicknesses and areas were lower in the seropositive group than in the other groups (p < 0.001, for all). CVI increased in both groups with "Bonzai" use compared to the control group (p < 0.001, for all). This study indicates that intraocular pressure may decrease, and choroidal changes may be observed in SC users. Further clinical studies with a larger sample size, especially using purified SC for therapeutic purposes, are needed to confirm the present findings, and further histopathologic studies are required to clarify the changes in the choroid despite SC seronegativity. NCT06235346.

Simultaneous determination of 102 synthetic cannabinoids in electronic cigarette oil by liquid chromatography-tandem mass spectrometry

Synthetic cannabinoids (SCs), which are among the most widely abused new psychoactive substances, are much more potent and have greater efficacy than natural cannabis. SCs can be disguised in various ways and are commonly sold in the form of electronic cigarette oil. SCs belong to a large family with structures consisting of a core with substituents, linker, ring with substituents, and tail. New SCs can be developed by adding substituents, such as halogen, alkyl, and alkoxy groups, to the aromatic ring system or by changing the alkyl chain length. Since the emergence of so-called first-generation SCs, subsequent developments have led to eighth-generation indole/indazole amide-based SCs. As of July 1, 2021, the entire category of SCs was added to the list of controlled substances, but implementation requires urgent improvements in detection technologies. Typically, each method is limited to a few SCs. Owing to the vast number of chemically diverse SCs and their fast update speed, the determination and identification of various types of SCs using a single method is challenging. Therefore, rapid, sensitive, and accurate quantitative methods that includes various types of SCs must be developed to meet the demand for the qualitative and quantitative analysis of new SCs in seized electronic cigarette oil. In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of 102 SCs in electronic cigarette oil. The mass spectrometry and liquid-phase conditions influencing SC separation and determination were optimized. Using the external standard method, 102 SCs were successfully identified in electronic cigarette oil. The samples were extracted using methanol. Target analytes were separated on a Shimadzu Shim-pack GIST-HP C18 AQ column (100 mm×2.1 mm, 1.9 μm) at a column temperature of 40 ℃. The mobile phases consisted of (A) 0.1% formic acid aqueous solution and (B) methanol-acetonitrile (1∶1, v/v). The gradient elution conditions were as follows: 0-8 min, 55%A-15%A; 8-15 min, 15%A; 15-16 min, 15%A-55%A; 16-18 min, 55%A. The flow rate was 0.4 mL/min and the injection volume was 1 μL. Operating in the multiple reaction monitoring mode, the 102 SCs were identified within 18 min. Each SC exhibited a good linear relationship in the range of 1-100.0 μg/L with a correlation coefficient (r)≥0.9915. The limits of detection were 0.01-0.30 μg/L and the limits of quantification were 0.04-0.99 μg/L, which meet the requirements for analyzing SCs in actual samples. Precision was determined using standard solutions with 2, 10, and 50 μg/L of the SCs. The precisions (n=6) were 0.3%-6.0%. The recoveries of the 102 SCs, as evaluated by spiking electronic cigarette oil at low (2 μg/mL), medium (10 μg/mL), and high (50 μg/mL) levels, were 80.1%-119.8%. Good performance was observed for the analysis of real samples. The developed method is accurate, rapid, sensitive, and effective for the determination of the 102 SCs in electronic cigarette oil, satisfying the requirements for practical qualitative and quantitative analysis.

Subacute Toxicity and Pharmacokinetic Evaluation of the Synthetic Cannabinoid 4F-MDMB-BUTINACA in Rats: A Forensic and Toxicological Perspective

Background: 4-MDMB-BUTINACA, a next-generation synthetic cannabinoid, presents significant public health and forensic challenges due to its evolving nature and potential toxicity. Methods: This study evaluates the subacute toxic effects and pharmacokinetics of 4−Fluoro MDMB−BUTINACA (4F-MDMB-BUTINACA) in adult male albino rats, administered orally for seven days at doses of 1 mg/kg, 5 mg/kg, and 15 mg/kg. The hematological, biochemical, and histopathological parameters were assessed and compared to controls. Results: The pharmacokinetics were determined using GC–MS/MS with a positive chemical ionization and granisetron as an internal standard. A histological analysis revealed inflammatory cell aggregation, congestion, hemorrhage, edema, and fibrosis in various tissues, with renal examinations showing tubule degradation, glomerular atrophy, Bowman’s space expansion, edema, and hemorrhage. The liver exhibited cellular infiltration, while cardiac muscle fibers showed myocardial fiber degradation and inflammatory cell aggregation. Biochemical assays indicated significant alterations (p &lt; 0.05) in the serum levels of AST, ALT, ALP, GGT, total protein, albumin, triglycerides, urea, MCHC, MCV, RDW, platelets, neutrophils, eosinophils, and basophils compared to the controls. Conclusions: The validated bioanalytical method revealed rapid absorption of 4F-MDMB-BUTINACA, with a plasma half-life of 2.371 h, a volume of distribution of 2272.85 L, and a plasma clearance rate of 664.241 L/h. In conclusion, 4F-MDMB-BUTINACA is a highly toxic synthetic cannabinoid, particularly affecting the liver, kidneys, and heart.

Classification of Mass Spectral Data to Assist in the Identification of Novel Synthetic Cannabinoids.

Detection and characterization of newly synthesized cannabinoids (NSCs) is challenging due to the lack of availability of reference standards and chemical data. In this study, a binary classification system was developed and validated using partial least square discriminant analysis (PLS-DA) by utilizing readily available mass spectral data of known drugs to assist in the identification of previously unknown NCSs. First, a binary classification model was developed to discriminate cannabinoids and cannabinoid-related compounds from other drug classes. Then, a classification model was developed to discriminate classical (THC-related) from synthetic cannabinoids. Additional models were developed based on the most abundant functional groups including core groups such as indole, indazole, azaindole, and naphthoylpyrrole, as well as head and tail groups including 4-fluorobenzyl (FUB) and 5-Fluoropentyl (5-F). The predictive ability of these models was tested via both cross-validation and external validation. The results show that all models developed are highly accurate. Additionally, latent variables (LVs) of each model provide useful mass to charge (m/z) for discrimination between classes, which further facilitates the identification of different functional groups of previously unknown drug molecules.

Exploring the Potential of Synthetic Cannabinoids: Modulation of Biological Activity of Normal and Cancerous Human Colon Epithelial Cells.

Colorectal cancer (CRC) is a global problem. Oncology currently practices conventional methods of treating this carcinoma, including surgery, chemotherapy, and radiotherapy. Unfortunately, their efficacy is low; hence, the exploration of new therapies is critical. Recently, many efforts have focused on developing safe and effective anticancer compounds. Some of them include cannabinoids. In the present study, we obtained cannabinoids, such as cannabidiol (CBD), abnormal cannabigerol (abn-CBG), cannabichromene (CBC), and cannabicitran (CBT), by chemical synthesis and performed the biological evaluation of their activity on colon cancer cells. In this study, we analyzed the effects of selected cannabinoids on the lifespan and metabolic activity of normal colonic epithelial cells and cancer colon cells. This study demonstrated that cannabinoids can induce apoptosis in cancer cells by modulating mitochondrial dehydrogenase activity and cellular membrane integrity. The tested cannabinoids also influenced cell cycle progression. We also investigated the antioxidant activity of cannabinoids and established a relationship between the type of cannabinoid and nitric oxide (NO) production in normal and cancerous colon cells. To conclude, it seems that, due to their interesting properties, the cannabinoids studied may constitute an interesting target for further research aimed at their use in alternative or combined therapies for human colon cancer.

Towards policy impact – an exploration of Clinical Toxicology research cited in policy documents and patents

Introduction Individual researcher impact through scientific citations is carefully monitored, with little attention to the impact of individual journals through policy and patent mentions. We aimed to describe policy and patent mentions for articles published in Clinical Toxicology. Methods Using Altmetric Explorer, we extracted mentions from 1 January 2013 to 31 December 2023, noting the citing source, Clinical Toxicology article title, and author-generated keywords. We used descriptive statistics to analyse the data. Results We identified 165 individual policy documents (n = 139) and patents (n = 26), citing 146 articles with median of 6.4 years between publication and mention. The highest number of citing documents were by the World Health Organization (n = 45), European Monitoring Centre for Drugs and Drug Addiction (n = 22), and United States Centers for Disease Control and Prevention (n = 16). Most patents were registered in the United States (n = 17) and by the European Patent Office (n = 10), with the main classification of human necessities (n = 23). The commonest subjects of papers cited in policy and patents, from keywords, related to medical conditions and symptoms (26%) and recreational drugs (22%). The most cited article was “A systematic review of adverse events arising from the use of synthetic cannabinoids and their associated treatment.” Discussion Clinical Toxicology articles are cited in policy documents and patents, with a comparable number of mentions to the top-ranked journals in the field. This likely contributes to policy impact, but further work is needed to understand how cited articles are used and ripple effects through onwards citations of policy documents. Conclusions Clinical Toxicology is a toxicology journal for which published research gets recognised within influential policy sources. The Journal can play a key role in guiding public health policy through its selection and development of submitted publications.

Difficulties in emotion regulation and attachment styles among Kurdish individuals in Eastern Turkey with substances use disorders

The aim of this study was to investigate the relationship between attachment style and emotion dysregulation in Kurdish individuals with substance use disorders (SUDs) in Eastern Turkey, a non-Western, Islamic society, in a descriptive cross-sectional design. This study was conducted with 216 individuals with SUDs who were treated at the SBU Van Training and Research Hospital Alcohol and Substance Addiction Treatment Center in Eastern Turkey between April 2023 and June 2023. Almost half of the participants (44.5%, n = 96) were between 18 and 30 years old and almost all were men (96.3%, n = 208). The most commonly used substances were heroin (46.3%, n = 100), marijuana (28.7%, n = 62) and synthetic cannabinoids (7.9%, n = 12). The results showed that higher levels of emotion dysregulation were associated with increased avoidant attachment and anxious attachment style. The study found that anxious and avoidant attachment styles were a significant predictor of emotion dysregulation. These findings suggest that attachment styles may play an important role in emotion dysregulation in Kurdish individuals with SUDs. Future research should investigate whether interventions targeting attachment-based interventions could be effective in reducing emotion dysregulation in Kurdish individuals with SUDs.

Bar adsorptive microextraction and liquid chromatography-diode array detection of synthetic cannabinoids in oral fluid.

In recent years, synthetic cannabinoids (SCs) have become a major public health issue. For this reason, there is a need for innovative analytical methods that allow its monitoring in biological matrices. In this work, we propose a novel methodology to screen eight SCs (AM-694, cumyl-5F-PINACA, MAM-2201, 5F-UR-144, JWH-018, JWH-122, UR-144 and APINACA) in oral fluids. A bar adsorptive microextraction method followed by microliquid desorption combined with high-performance liquid chromatography with diode array detection (BAµE-µLD/HPLC-DAD) was developed to monitor the target SCs. The main factors affecting the BAµE technology were fully optimized for oral fluid analysis. Under optimized experimental conditions, the proposed methodology showed good linear dynamic ranges from 20.0 to 2000.0 µg L-1 (r2 > 0.99, relative residuals < 15%), limits of detection between 2.0 and 5.0 µg L-1 and suitable average recovery yields (87.9-100.5%) for the eight studied SCs. The intra- and interday accuracies (bias ≤ ± 14.7%) and precisions (RSD ≤ 14.9%) were also evaluated at three spiking levels. The validated methodology was then assayed to oral fluid samples collected from several volunteers. The proposed analytical approach showed remarkable performance and could be an effective alternative for routine monitoring of the target compounds in oral fluid.

Detection and quantification of synthetic cannabinoids in seven illicitly sourced disposable vapes submitted by an individual presenting to a UK drug and alcohol service.

In the United Kingdom and internationally, synthetic cannabinoids (SCs) are a common adulterant in illicitly sourced vaping products. Recently, their use is increasingly being linked to severe health effects, particularly among children. Here, we aimed to conduct the first detection and quantification of SCs in illicit disposable vaping products. A cross-section of seven illicitly sourced disposable vape samples that were initially sold as cannabis products was submitted for analysis by a single individual presenting to a drug and alcohol service in the United Kingdom. Qualitative and quantitative analyses of these samples were conducted using nuclear magnetic resonance and gas chromatography/electron ionization-mass spectrometry. Qualitative analysis identified the SC 5F-MDMB-PICA in all seven samples, in the absence of any other pharmacologically active compounds. Quantitative analysis revealed that the median concentration of 5F-MDMB-PICA was 0.85 mg/ml (range = 0.59-1.63). The external appearance of these vape samples closely resembled regulated vaping products, and the presence of SCs was not identifiable by any labelling or packaging. The SC 5F-MDMB-PICA was detected at a median concentration of 0.85 mg/ml in seven disposable vapes which were illegally sourced in the United Kingdom, were mis-sold as cannabis products and closely resembled legal, regulated products.

Development and method validation of a sampling technique for a reproducible detection of synthetic cannabinoids in exhaled breath using an in vitro pig lung model.

Alternative matrices, especially exhaled breath (EB), have gained increasing attention for a few years. To interpret toxicological findings, knowledge on the toxicokinetic (TK) properties of a substance in EB is indispensable. Whilst such data are already accessible for various drugs (e.g. Δ9-tetrahydrocannabinol), they are still not available for new psychoactive substances, particularly synthetic cannabinoids (SCs). As SCs raise a high public health concern, the aim of this study was to assess these data in future TK studies in pigs. For this purpose, an in vitro sampling technique of EB was initially developed, being prospectively applied to anesthetized and ventilated pigs for the detection of SCs in a controlled and reproducible manner as exemplified by cumyl-5F-P7AICA. Furthermore, a method for the qualitative and quantitative detection of cumyl-5F-P7AICA in EB using glass fiber filters (GFF) was established und fully validated. Therefore, cumyl-5F-P7AICA (0.5 mg/mL in ethanol abs.) was initially nebulized using a ventilation machine and a breathing tube, as they are also used in surgeries. The aerosol was delivered into a simulated pig lung. To collect EB, a pump was connected to that part of the breathing tube, that contains EB (expiratory limb), and sampling was performed repeatedly (n=6) for 15 min (2 L EB/min) each using GFF. For extraction of the substance, the GFF were macerated with acetone and the remaining experimental components were rinsed with ethanol. After sample preparation, the extracts were analyzed by LC-MS/MS. In the complete experimental setup, about 40% of the initially nebulized cumyl-5F-P7AICA dose was found with 3.6 ± 1.3% being detected in the GFF. Regarding the comparably high loss of substance, the open ventilation system and a conceivable adsorption of the SC in the ventilator have to be considered. However, the herein introduced approach is promising to determine the TK properties of cumyl-5F-P7AICA in EB.

Exploiting the triple quadrupole mass analyzer for the open detection and tentative identification of synthetic cannabinoid receptor agonists based on common fragmentation pathways

BackgroundThe use of new psychoactive substances (NPS) has emerged as a significant public health concern globally, due to their unknown and unpredictable effects on both physical and mental health. Among them, synthetic cannabinoids receptor agonists (SCRAs) currently stand as the most widely consumed NPS family in Europe. Since the detection of JWH-018 in 2008, the structures of these compounds have evolved to circumvent legislation and/or enhance their effects, consequently increasing the number of reported SCRAs to be monitored. Therefore, new strategies are needed to identify these compounds, whether in seized products or in biological samples. ResultsThis study presents the development of an open method for detecting SCRAs employing a “pseudo-target” screening approach, a strategy previously developed and used in our laboratory for synthetic cathinones identification. The methodology involves monitoring the main product ions and neutral losses derived from 179 SCRAs of the third and fourth generations, based on their fragmentation pathways. This approach allows for the tentative identification of the SCRAs, supported also by the created database. The versatility of the developed methodology is highlighted, extending its utility beyond seizure products or ‘legal highs’, to biological samples. In this sense, it has been successfully applied not only to the detection of SCRAs in research chemicals but also in authentic urine from an anonymous SCRAs consumer, through the identification of a metabolite. SignificanceThis strategy will be particularly useful for the rapid detection of SCRAs in forensic and toxicological laboratories equipped with low-resolution MS/MS instrumentation. This is a valuable tool for the identification and monitoring of SCRAs across various contexts, significantly contributing to public health and forensic security efforts. It is especially beneficial for healthcare providers, enabling them to make informed treatment decisions.

Mechanistic insights into the impact of WIN 55, 212-2, a synthetic cannabinoid, on adhesion molecules PECAM-1 and VE-cadherin in HeLa cells: implications on cancer processes

The endocannabinoid (eCB) system comprises endogenous ligands, cannabinoid receptors (CBRs), and their regulatory proteins; its alteration leads to many diseases including cancer. Thus, becomes a therapeutic target for synthetic cannabinoids aimed to control cancer cell proliferation, migration, adhesion, and invasion. However, little is known about adhesion molecules regulation through CBRs activation. The aim of this study was to evaluate the effects of a CB1/CB2 agonist, WIN-55, 212-2 (WIN), on the regulation of adhesion molecules platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial cadherin (VE-cadherin) in HeLa cells. CBRs expression was evaluated by immunofluorescence staining in HeLa cells and cell viability (thiazolyl blue tetrazolium bromide), cell adhesion (crystal violet), adhesion molecules expression and location (Western blot and immunofluorescence staining assays) were all assessed on cells treated with different WIN concentrations. Receptors CB1, CB2, and G-protein-coupled receptor 55 were expressed in HeLa cells. Additionally, biphasic effects were observed in their metabolic activity and adhesive properties: low WIN concentrations resulted in significant increases whereas, high ones decreased them compared to controls (p < 0.0001), demonstrating that WIN elicits opposite effects depending on the concentration and exposure time. PECAM-1 was detected in HeLa cell’s cytoplasm, membrane, and perinuclear region, whereas VE-cadherin had a nuclear distribution. There were no significant differences in PECAM-1 and VE-cadherin expression and location, suggesting that WIN does not modulate these proteins. These findings support the potential use of WIN due to its anticancer properties without dysregulating adhesion molecules. WIN possible contribution to inhibit cancer progression should be further investigated.

The Impact of Potent Addictive Substances on Angiogenic Behavior: A Comprehensive Review.

Angiogenesis, the formation of new vasculature from preexisting vasculature, is involved in the development of several diseases as well as various physiological processes. Strict cooperation of proangiogenic and antiangiogenic factors mediates the control of angiogenesis. The fundamental steps in angiogenesis include endothelial cell proliferation, migration, and invasion. Addictive substances, which are considered therapeutic candidates in research and medicine, are classified as natural substances, such as nicotine, or synthetic substances, such as synthetic cannabinoids. Addictive substances have been shown to either enhance or suppress angiogenesis. This review article provides an overview of recent studies concerning the effects of several addictive substances on the process of angiogenesis. Google Scholar and PubMed were used to collect the scientific literature used in this review. The addictive substances addressed in this review are nicotine, opioids such as morphine and heroin, alcohol, cocaine, methamphetamine, and cannabinoids. An accurate assessment of the influence of these substances on the angiogenic process may help to construct a potentially effective therapeutic protocol to control and treat several angiogenesis-related diseases.