Prognostic value of the expression and localization of cell proliferation and apoptosis markers in unicystic ameloblastomas.

The aim of this study was to verify whether the expression of cell proliferation and apoptosis markers in different types of unicystic ameloblastoma (UA) is associated with the location of neoplastic cells. Immunohistochemical study with a sample of 32 cases of UA, 11 cases of conventional ameloblastoma (CAM) and ten dental follicles (DF) cases was performed. Cell proliferation was assessed using Ki-67 status, and apoptosis by caspase-3 expression. Mural UA (MUA) showed a higher immunostaining of Ki-67 (p < 0.05) and a lower immunostaining of Caspase-3 (p < 0.05) compared with luminal and intraluminal subtypes of UA and CAM. The neoplastic cells of the MUA's cystic capsule showed a higher expression of Ki-67 protein (p < 0.0001) and a lower expression of Caspase-3 (p < 0.0001) compared with the lumen. DF showed lower Ki-67 and Caspase-3 immunostaining (p < 0.05) than neoplasms. The higher immunoexpression of Ki-67 and the lower immunoexpression of Caspase-3 in MUA, in the parenchyma cells within the cystic capsule, suggest an association between the biological behaviour and location of neoplastic cells in a tumour.

Interactive Effects of Ayahuasca and Cannabidiol in Social Cognition in Healthy Volunteers: A Pilot, Proof-of-Concept, Feasibility, Randomized-Controlled Trial.

BACKGROUND: Serotonergic hallucinogens and cannabinoids may alter the recognition of emotions in facial expressions (REFE). Cannabidiol (CBD) attenuates the psychoactive effects of the cannabinoid-1 agonist tetrahydrocannabinol. Ayahuasca is a dimethyltryptamine-containing hallucinogenic decoction. It is unknown if CBD may moderate and attenuate the effects of ayahuasca on REFE. PROCEDURES: Seventeen healthy volunteers participated in a 1-week preliminary parallel-arm, randomized controlled trial for 18 months. Volunteers received a placebo or 600 mg of oral CBD followed by oral ayahuasca (1 mL/kg) 90 minutes later. Primary outcomes included REFE and empathy tasks (coprimary outcome). Tasks were performed at baseline and 6.5 hours, 1 and 7 days after the interventions. Secondary outcome measures included subjective effects, tolerability, and biochemical assessments. RESULTS: Significant reductions (all P values <0.05) only in reaction times were observed in the 2 tasks in both groups, without between-group differences. Furthermore, significant reductions in anxiety, sedation, cognitive deterioration, and discomfort were observed in both groups, without between-group differences. Ayahuasca, with or without CBD, was well tolerated, producing mainly nausea and gastrointestinal discomfort. No clinically significant effects were observed on cardiovascular measurements and liver enzymes. CONCLUSIONS: There was no evidence of interactive effects between ayahuasca and CBD. The safety of separate and concomitant drug intake suggests that both drugs could be applied to clinical populations with anxiety disorders and in further trials with larger samples to confirm findings.

Avaliação da completude dos dados do sistema de informação sobre nascidos vivos de imigrantes e brasileiros no estado do Paraná, 2014-2019 / Evaluation of the completeness of data in the information system on live births of immigrants and brazilians in the state of Paraná, 2014-2019 / Evaluación de la completitud de los datos del sistema de información sobre nacidos vivos de inmigrantes y brasileños en el estado de Paraná, 2014-2019

Avaliar a completude dos dados no SINASC do Paraná, entre 2014 a 2019, de imigrantes e brasileiros. Método: Estudo transversal, retrospectivo, de análise de completude dos dados de nascimento do ano de 2014 a 2019 no estado do Paraná. Foram critérios de análise: excelente (menos de 5% de incompletude); bom (5% a 10%); regular (10% a 20%); ruim (20% a 50%); e muito ruim (acima de 50%). Resultados: Foram registrados no Brasil 948.316 nascidos vivos, dos quais 935.629 eram de mães brasileiras e 12.867 de mães imigrantes de diversas nacionalidades. Dentre as variáveis, os campos incompletos de mães brasileiras somaram 50.243 (5,37%) e de imigrantes 696 (5,41%), demonstrando um bom preenchimento do banco. Conclusão: Foi possível verificar que o banco de dados SINASC mostrou-se confiável e com baixas incompletudes entre os anos de 2014 a 2019 no estado do Paraná, independente da naturalidade da mãe. Porém, dados incompletos referentes às imigrantes ainda são maiores comparados aos de mulheres brasileiras, podendo ser resultado de uma falta de capacitação dos profissionais da saúde para a comunicação com as mães imigrantes que não falam a língua nativa, não coletando os dados de forma adequada e completa.

To evaluate the completeness of data in the SINASC of Paraná, from 2014 to 2019, of immigrants and Brazilians. Method: Cross-sectional, retrospective study, of analysis of completeness of birth data from the year 2014 to 2019 in the state of Paraná. Analysis criteria were: excellent (less than 5% incompleteness); good (5% to 10%); fair (10% to 20%); bad (20% to 50%); and very bad (above 50%). Results: In Brazil, 948,316 live births were registered, of which 935,629 were from Brazilian mothers and 12,867 from immigrant mothers of various nationalities. Among the variables, the incomplete fields of Brazilian mothers totaled 50,243 (5.37%) and of immigrants 696 (5.41%), de- monstrating a good completion of the bank. Conclusion: It was possible to verify that the SINASC database proved to be reliable and with low incompleteness between the years 2014 to 2019 in the state of Paraná, regardless of the mother's naturality. However, in- complete data referring to immigrants are still higher compared to those of Brazilian wo- men, which may be the result of a lack of training of health professionals for communi- cation with immigrant mothers who do not speak the native language, not collecting the data properly and completely.

Evaluar la completitud de datos en el SINASC de Paraná, de 2014 a 2019, de inmigrantes y brasileños. Método: Estudio transversal, retrospectivo, de análisis de completitud de datos de nacimiento del año 2014 a 2019 en el estado de Paraná. Los criterios de análisis fueron: excelente (menos de 5% de incompletitud); bueno (5% a 10%); regular (10% a 20%); malo (20% a 50%); y muy malo (más de 50%). Resultados: En Brasil se registraron 948.316 nacidos vivos, de los cuales 935.629 eran de madres brasileñas y 12.867 de madres inmigrantes de diversas nacionalidades. Dentre as variá- veis, os campos incompletos de mães brasileiras somaram 50.243 (5,37%) e de imigrantes 696 (5,41%), demonstrando um bom preenchimento do banco. Conclusão: Foi possível verificar que o banco de dados SINASC mostrou-se confiável e com baixa incompletudes entre os anos de 2014 a 2019 no estado do Paraná, independentemente da naturalidade da mãe. Sin embargo, los datos incompletos referidos a las inmigrantes siguen siendo más altos en comparación con los de las mujeres brasileñas, lo que puede ser el resultado de la falta de formación de los profesionales de la salud para la comunicación con las madres inmigrantes que no hablan el idioma nativo, no recogiendo los datos de forma adecuada y completa.

Maintenance protocols for implant-supported dental prostheses: A scoping review.

STATEMENT OF THE PROBLEM: Biological complications of implant-supported prostheses remain a concern. Therefore, standardizing hygiene protocols to promote their maintenance is important. PURPOSE: The purpose of this scoping review was to identify available hygiene guidance for home care procedures, as well as periodicity and protocols for the professional maintenance of implant-supported prostheses. MATERIAL AND METHODS: This study was based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, and the study protocol was made available at: https://osf.io/5jdeh/. The search was last performed in the PubMed database on March 15, 2022 and was undertaken by 2 independent researchers. Clinical studies and reviews that evaluated home care and maintenance protocols for dental implant-supported prostheses (fixed or removable), without language restriction or year of publication, were included. A descriptive analysis was performed considering study characteristics (study design, restorative assembly, maintenance strategies, eligibility criteria, number of included studies, quality/level of evidence, and main findings). RESULTS: The initial search yielded 3138 studies, of which 18 were included for descriptive analysis (6 critical reviews, 4 clinical trials, 3 systematic reviews, 2 guideline reports, 2 retrospective studies, and 1 transversal study). The main instrument recommended for home care was the use of a conventional toothbrush associated with triclosan-containing toothpaste in addition to interproximal aids (brushes or floss) for all types of restorations. The use of irrigation instruments was also frequently considered. For professional maintenance, almost all studies reported a positive effect of regular recalls every 3 months during the first year, followed by less regular recalls according to the motivation of the patient and home care efficacy. At each recall, the professional should evaluate the patient history, oral tissues, implant, abutments, and restorations, as well as perform professional cleaning of the prostheses with appropriate instruments so that the implants and abutments may be preserved. CONCLUSIONS: Establishing an adequate hygiene protocol is indispensable for implant-supported restorations. To do so, the professional must guide and support the patient's home care by taking into account the patient's motivation and efficacy during the hygiene procedures. Also, for professional maintenance, the periodicity of recalls should be constant and include an evaluation of the condition of the restoration and adjacent tissues, followed by professional cleaning of prostheses, implants, and abutments, in addition to new instructions to improve patient home care.

Analgesia and Side Effects of Codeine Phosphate Associated with Paracetamol Versus Oxycodone After the Extraction of Mandibular Third Molars: A Randomized Double-Blind Clinical Trial Using the Split-Mouth Model.

Purpose: To assess the analgesia and side effects of 10 mg oxycodone as compared to 30 mg of codeine phosphate associated with 500 mg of paracetamol after bilateral lower third molar extraction. Methods: This is a prospective, randomized, double-blind study applied to a sample of 16 patients. They were evaluated for seven days postoperatively, and the mean score of the visual analogue scale (VAS) of pain between test and control medications was assessed by the Wilcoxon distribution. The side effects of these medications were assessed by the Q Cochran test. A p value of < .05 was considered statistically significant. Results: The mean score of the VAS of pain was higher in the oxycodone side, where few patients reported the use of rescue analgesic. There was no report of rescue medication in codeine phosphate associated with paracetamol side. The most common side effects reported in both groups, predominantly in patients using the oxycodone, were drowsiness, dizziness, and headache. Conclusion: The use of codeine phosphate associated with paracetamol after the extraction of impacted mandibular third molars is a better choice than oxycodone for controlling postoperative pain. Trial Registration Number and Date of Registration: RBR-8ntwmyq 07/07/2021.

Evidence on the impairing effects of Ayahuasca on fear memory reconsolidation.

RATIONALE: To uncover whether psychedelic drugs attenuate fear memory responses would advance the development of better psychedelic-based treatments for posttraumatic stress disorder (PTSD). Ayahuasca (AYA), a psychedelic brew containing indolamine N, N-dimethyltryptamine (DMT) and ß-carbolines, facilitates fear extinction and improves neural plasticity. Upon retrieval, fear memory undergoes labilization and reconsolidation; however, the effects of AYA on this memory stabilization phase are unknown. OBJECTIVES: We aimed to investigate the effects of AYA treatment on fear memory reconsolidation. METHODS: Fear-conditioned Wistar rats received AYA (60, 120, or 240 mg/kg) or H2O orally via gavage o.g. 20 min before, immediately, or 3 h after a short retrieval session. Analysis of AYA through liquid chromatography-tandem mass spectrometry was used to determine the content of DMT and ß-carbolines in AYA. RESULTS: AYA impaired fear memory reconsolidation when given 20 min before or 3 h after memory retrieval, with the dose of 60 mg/kg being effective at both moments. This dose of AYA was devoid of anxiolytic effect. Importantly, during retrieval, AYA did not change fear expression. The lack of retrieval abolished the reconsolidation impairing effect of AYA. The effects of AYA treatment 20 min before or 3 h after memory retrieval lasted at least 22 days, suggesting no spontaneous recovery of fear memory. Fear memory impairments induced by AYA treatment, at both moments, do not show reinstatement. CONCLUSIONS: Our findings support the view that a low dose of AYA treatment impairs early and late stages of memory reconsolidation instead of facilitating fear extinction.

Knowledge about oral cancer among dental students and Primary Health Care dentists: A Brazilian study.

PURPOSE/OBJECTIVES: The present study aims to evaluate the knowledge of dental students and dentists who work in Primary Health Care (PHC). These are professionals who work in the public basic health units regarding oral cancer prevention, diagnosis, and treatment. METHODS: A cross-sectional and observational Brazilian study was conducted. A survey with 27 questions was carried out regarding the (i) demographic, academic, and professional data of the participants, (ii) attitudes, practices, and self-perception about their training, and (iii) knowledge about oral cancer. With the exception of one question regarding the clinical characteristics of oral cancer, all knowledge answers were graded as correct or incorrect. Grades were assigned to each participant according to their level of knowledge, and scores for each participant were calculated ranging from 1 to 10. Mann-Whitney and Kruskal-Wallis tests were used to examine the difference in the knowledge score of oral cancer according to explanatory variables. The level of significance adopted was 5%. RESULTS: The sample comprised 541 dental students and 82 PHC dentists. Most participants did not consider it appropriate or had not received training for palpation of lymph nodes or for identifying lymphadenopathies associated with oral cancer. Less than 20% of the participants had some kind of practice before and felt safe performing biopsy surgery. The average student score was 7.01 ± 2.15, and the average dentist score was 7.81 ± 1.467. CONCLUSION: Although the scores were satisfactory, most of the participants did not show adequate knowledge about biopsy and palpation of cervical lymph nodes.

Effects of ayahuasca on the endocannabinoid system of healthy volunteers and in volunteers with social anxiety disorder: Results from two pilot, proof-of-concept, randomized, placebo-controlled trials.

OBJECTIVE: To assess endocannabinoid (anandamide, AEA; 2-arachidonoylglycerol, 2-AG) plasma levels in healthy volunteers and in volunteers with social anxiety disorder (SAD) after a single oral dose of ayahuasca or placebo. METHODS: Post hoc analysis of endocannabinoid plasma levels (baseline, 90 and 240 min after drug intake) from two parallel-group, randomized, placebo-controlled trials. In Study 1, 20 healthy volunteers ingested ayahuasca (average 1.58 mg/ml dimethyltryptamine (DMT)) or placebo, and in Study 2, 17 volunteers with SAD received ayahuasca (average 0.680 mg/ml DMT) or placebo. RESULTS: A significant difference was observed in AEA concentrations in Study 2 after ayahuasca intake (Χ2 (2) = 6.5, p = 0.03, Friedman test), and near significant differences (increases) were observed between baseline and 90 (Z = 0, p = 0.06, Wilcoxon test) and 240 (Z = 10, p = 0.06) minutes after ayahuasca intake. CONCLUSIONS: Although our findings suggest that ayahuasca could modulate AEA levels in SAD patients, the high interindividual variability in both trials and the small samples preclude definitive conclusions. More research with larger samples is needed to better understand the effects of ayahuasca and other hallucinogens in the endocannabinoid system.

Ayahuasca, a psychedelic beverage, modulates neuroplasticity induced by ethanol in mice.

Alcohol use disorder needs more effective treatments because relapse rates remain high. Psychedelics, such as ayahuasca, have been used to treat substance use disorders. Our study aimed to evaluate the effects of ayahuasca on ethanol-induced behavioral sensitization (EIBS). Swiss mice received 2.2 g/kg ethanol or saline IP injections every other day across nine days (D1, D3, D5, D7, and D9), and locomotor activity was evaluated 10 min after each injection. Then, animals were treated daily with ayahuasca (corresponding to 1.76 mg/kg of N,N-dimethyltryptamine, DMT) or water by oral gavage for eight consecutive days. On the seventh day, mice were evaluated in the elevated plus maze. Then, mice were challenged with a single dose of ethanol to measure their locomotor activity. Dopamine receptors, serotonin receptors, dynorphin, and prodynorphin levels were quantified in the striatum and hippocampus by blot analysis. Repeated ethanol administration resulted in EIBS. However, those animals treated with ayahuasca had an attenuated EIBS. Moreover, ayahuasca reduced the anxiogenic response to ethanol withdrawal and prevented the ethanol-induced changes on 5-HT1a receptor and prodynorphin levels in the hippocampus and reduced ethanol effects in the dynorphin/prodynorphin ratio levels in the striatum. These results suggest a potential application of ayahuasca to modulate the neuroplastic changes induced by ethanol.

Effects of Ayahuasca on Personality: Results of Two Randomized, Placebo-Controlled Trials in Healthy Volunteers.

Rationale: Previous studies with the serotonergic hallucinogens LSD and psilocybin showed that these drugs induced changes in personality traits, such as increases in Openness. However, results are inconsistent, and the effects of ayahuasca on personality were never investigated in a controlled trial. Objectives: To assess the effects of ayahuasca on personality in two randomized, placebo-controlled trials in healthy volunteers. Methods: Data from two parallel-group, randomized, placebo-controlled trials in healthy volunteers were included. In the first trial, 15 volunteers ingested ayahuasca or placebo, while in the second trial 15 volunteers received placebo+ayahuasca or cannabidiol (CBD)+ayahuasca. Personality was assessed with the NEO-Five Factor Inventory (NEO-FFI) at baseline and 21 days post-treatment. Results: There were significant differences between groups in baseline Openness scores, but not on day 21. A significant increase in Openness scores was observed in the placebo + ayahuasca group in study 2. No other within-group differences were observed for any other domain. Conclusions: Ayahuasca produced inconsistent effects on personality since it induced significant increase in Openness 21 days post-drug intake only in one of the trials. The absence of significant differences in the other ayahuasca groups could be due to small sample sizes and baseline differences among groups. The effects of ayahuasca and other serotonergic hallucinogens on personality should be further investigated in clinical samples.

Ayahuasca Improves Self-perception of Speech Performance in Subjects With Social Anxiety Disorder: A Pilot, Proof-of-Concept, Randomized, Placebo-Controlled Trial.

BACKGROUND: Ayahuasca is a classic hallucinogen with anxiolytic and antidepressive properties. Anecdotal evidence also suggests that it improves performance (eg, singing, speech). This controlled trial assessed the effects of ayahuasca on speech performance and anxiety in individuals with social anxiety disorder. METHODS: Seventeen volunteers with social anxiety disorder participated in a pilot, proof-of-concept, randomized, parallel-group trial. Self-perception of performance during a public-speaking test was assessed with the Self-statements During Public Speaking Scale primary outcome). Secondary outcomes included anxiety/subjective effects (Visual Analog Mood Scale; Bodily Symptoms Scale), recognition of emotions in facial expressions (REFE), tolerability measures (cardiovascular measures, self-reports), and brain-derived neurotrophic factor plasma levels. Effects on anxiety and REFE were assessed again 7, 14, and 21 days postdrug. FINDINGS: Compared with placebo, ayahuasca significantly improved self-perception of speech performance (Self-statements During Public Speaking Scale) and increased somatic symptoms (Bodily Symptoms Scale). There was also a significant time × group interaction in the cognitive deterioration Visual Analog Mood Scale factor and a significant effect of time in the REFE task, especially in reaction time. Other measures were not significantly modified. Ayahuasca was well tolerated, producing mainly nausea, gastrointestinal discomfort, and vomiting. CONCLUSIONS: Ayahuasca improved self-perception of speech performance in socially anxious individuals. These effects occurred independent of task-related anxiety and REFE, suggesting that ayahuasca could specifically improve the cognitive aspect of speech performance. Further studies should try to unveil the mechanisms involved in the effects of ayahuasca and to better understand its effects on anxiety.

Application of Construction and Demolition Waste in Civil Construction in the Brazilian Amazon-Case Study of the City of Rio Branco.

The lack of usable aggregates for civil construction in Rio Branco (capital of Acre, a Federal State in the Amazon region) makes the production and use of recycled aggregates from construction and demolition waste (CDW) an alternative of great interest. In this study, a comprehensive characterization of CDW collected from 24 construction sites of six building types and three different construction phases (structures, masonry, and finishing) was carried out. The fine and coarse recycled aggregates were produced and evaluated in 10 different compositions. The aggregates' performance was evaluated in four mixtures designed for laying and coating mortars with a total replacement of conventional aggregates and a mixture designed for a C25 concrete with 50% and 100% replacement of conventional aggregates. CDW mortars showed lower densities and greater water retention, initial adhesion, and mechanical strength than conventional mortars. CDW concretes presented lower densities and greater resistance to chloride penetration than conventional concrete, with a small mechanical strength reduction. The recycled CDW aggregates proved to be technologically feasible for safe application in mortars and concrete; for this reason, it is believed that the alternative and proposed methodology is of great interest to the Amazonian construction industry, considering the high costs of raw materials and the need for defining and consolidating a sustainable development model for the Amazon region.

Effects of Ayahuasca on the Recognition of Facial Expressions of Emotions in Naive Healthy Volunteers: A Pilot, Proof-of-Concept, Randomized Controlled Trial.

BACKGROUND: The recognition of emotions in facial expressions (REFE) is a core aspect of social cognition. Previous studies with the serotonergic hallucinogens lysergic acid diethylamide and psilocybin showed that these drugs reduced the recognition of negative (fear) faces in healthy volunteers. This trial assessed the acute and prolonged effects of a single dose of ayahuasca on the REFE. METHODS: Twenty-two healthy volunteers participated in a pilot, proof-of-concept, randomized trial. Study variables included a REFE task performed before and 4 hours after drug intake, subjective effects (self-reports/observer impressions), tolerability measures (cardiovascular measures, self-reports), and brain-derived neurotrophic factor plasma levels. The REFE task was applied again 1, 7, 14, and 21 days and 3 months after drug intake. Stability of ayahuasca alkaloids during the study was also assessed (room temperature, 18 months). FINDINGS: Compared with placebo, ayahuasca did not modify the REFE. No significant effects were observed on cardiovascular measures and brain-derived neurotrophic factor levels. Volunteers reported visual effects, tranquility/relaxation, and well-being, with few reports of transient anxiety/confusion. Ayahuasca was well tolerated, producing mainly nausea, gastrointestinal discomfort, and vomiting. A significant time-dependent deterioration of alkaloids was observed, especially for dimethyltryptamine. CONCLUSIONS: Absence of significant effects on the REFE task could be due to lack of effects of ayahuasca (at the doses used), alkaloid degradation, learning effects, and the high educational level of the sample. Further trials with different samples are needed to better understand the effects of ayahuasca and other serotonergic hallucinogens on the REFE. Future trials should improve methods to guarantee the stability of ayahuasca alkaloids.

Ayahuasca prevents oxidative stress in a rat model of depression elicited by unpredictable chronic mild stress

ABSTRACT Background: Depression is a highly disabling common mental disorder and, due to its multifactorial nature, the development of effective therapies is challenging and of great relevance. Ayahuasca (AYA), an entheogenic traditional beverage, has emerged as an alternative for antidepressant treatment, however, AYA preclinical and clinical trials are still incipient. Objectives: This investigation aimed to evaluate some behavioral and biochemical effects of AYA subchronic administration in rats submitted to a model of depression elicited by unpredictable chronic mild stress (UCMS). Methods: 500 mg/kg of AYA was administered in adult male rats once a day for 15 days before submitting the animals to UCMS. Anhedonia-like and locomotion behavior, lipid peroxidation, antioxidant enzyme activities, and sulfhydryl/nitrite content were evaluated. Results: AYA intake failed to prevent anhedonia-like behavior. Locomotion was not altered by AYA consumption or by the experimental condition. UCMS increased TBARS and nitrites levels, decreased the levels of catalase in the cerebral cortex and of Sulfhydryl in the hippocampus. AYA treatment counteracted these biochemical alterations but did not display any alterations in non-stressed rats. Conclusions: Taken together, results indicate an adaptogenic antioxidant molecular mechanism of AYA in relation to depression induced by stress.

Essential oil-based dispersive liquid-liquid microextraction for the determination of N,N-dimethyltryptamine and ß-carbolines in human plasma: A novel solvent-free alternative.

The present study describes the development of a novel solvent-free vortex-assisted dispersive liquid-liquid microextraction alternative based on a natural essential oil as extracting solvent (VA-EO-DLLME) for the determination of N,N-dimethyltryptamine (DMT), harmine (HRM), harmaline (HRL) and tetrahydroarmine (THH) (compounds found in the ayahuasca tea, a psychedelic plant preparation) in human plasma. After optimization through full factorial and Box-Behnken experimental designs, this VA-EO-DLLME followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was completely validated and applied to authentic plasma specimens. Sample preparation consisted in the addition of 60 mg of NaCl, 100 µL of borate buffer and 100 µL of Eucalyptus globulus essential oil to a 200 µL aliquot of human plasma. After 30 s of vortex agitation followed by 5 min of centrifugation (10,000 rpm), 80 µL of the oil supernatant was dried and resuspended in mobile phase prior to injection into the UHPLC-MS/MS system. Once optimized, the validated method yielded LoDs ≤1.0 ng mL-1 for all analytes. LoQ was 1.0 ng mL-1 for DMT, HRL and HRM and 2.0 ng mL-1 for THH. The method has shown to be linear over the range of LoQ up to 150 ng mL-1 (r2 ≥ 0.9926). Intra/inter-day precision and accuracy met the acceptance criteria at three quality control (QC) levels. An additional intermediate precision study demonstrated that, except for THH and HRL at low and medium QCs, the overall method performance was similar for the three different oil sources. Matrix effect evaluation showed predominant ion suppression, ranging from 56% to 83%. Recovery varied from 33 up to 101% with an average of 50 ± 15.8%. Selectivity studies showed no interferences. Analysis of 13 authentic samples proved method feasibility. Finally, we believe that our novel VA-EO-DLLME approach offers a very simple, fast, cost-effective and eco-friendly alternative based on the use of an easily accessible and entirely green material as an extracting solvent. This may represent an incentive for researchers to investigate novel and creative alternatives, such as essential oils, as substitutes of organic solvents for microextraction methods in forensic and clinical contexts.

Kinetic profile of N,N-dimethyltryptamine and ß-carbolines in saliva and serum after oral administration of ayahuasca in a religious context.

Ayahuasca is a beverage obtained from Banisteriopsis caapi plus Psychotria viridis. B. caapi contains the ß-carbolines harmine, harmaline, and tetrahydroharmine that are monoamine oxidase inhibitors and P. viridis contains N,N-dimethyltryptamine (DMT) that is responsible for the visionary effects of the beverage. Ayahuasca use is becoming a global phenomenon, and the recreational use of DMT and similar alkaloids has also increased in recent years; such uncontrolled use can lead to severe intoxications. In this investigation, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to study the kinetics of alkaloids over a 24 h period in saliva and serum of 14 volunteers who consumed ayahuasca twice a month in a religious context. We compared the area under the curve (AUC), maximum concentration (Cmax ), time to reach Cmax (Tmax ), mean residence time (MRT), and half-life (t1/2 ), as well as the serum/saliva ratios of these parameters. DMT and ß-carboline concentrations (Cmax ) and AUC were higher in saliva than in serum and the MRT was 1.5-3.0 times higher in serum. A generalized estimation equations (GEEs) model suggested that serum concentrations could be predicted by saliva concentrations, despite large individual variability in the saliva and serum alkaloid concentrations. The possibility of using saliva as a biological matrix to detect DMT, ß-carbolines, and their derivatives is very interesting because it allows fast noninvasive sample collection and could be useful for detecting similar alkaloids used recreationally that have considerable potential for intoxication.

Fast Hollow Fiber Liquid-Phase Microextraction as a Greener Alternative for the Determination of N,N-Dimethyltryptamine and Harmala Alkaloids in Human Urine.

Ayahuasca tea is an entheogen hallucinogenic beverage used for shamanic and spiritual purposes, prepared by the decoction of different Amazonian plants containing N,N-dimethyltryptamine (DMT) and harmala alkaloids. Since the therapeutic potential of this tea has been broadly studied in recent years, mainly for the treatment of psychiatric disorders, the determination of the ayahuasca tea components in human and animal matrices is of utmost importance. In order to avoid the use of large amounts of toxic solvents, typically employed in traditional sample preparation methods, hollow fiber liquid-phase microextraction (HF-LPME) presents a greener and time-saving alternative. The present study aims to fully develop and apply an HF-LPME method for the determination of DMT, harmine (HRM), harmaline (HRL), and tetrahydroharmine (THH) in human urine samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fractional factorial and Box-Behnken designs were used to identify and optimize significant method variables. Once optimized, validation has shown a limit of detection (LoD) of 1.0 ng/ml for DMT and 2.0 ng/ml for the harmala alkaloid. The limit of quantification (LoQ) was of 5.0 ng/ml for all analytes. The method has shown to be linear over a concentration range of 5-200 ng/ml (r 2 ≥ 0.99). Intra/inter-day precision and accuracy met the acceptance criteria at the three quality control (QC) levels studied (15.0, 90.0, and 170.0 ng/ml, n = 6, each). Matrix effect evaluation showed predominant ion enhancement and recovery values were above 80%. Dilution factors of 10- and 20-fold have shown acceptable values of accuracy. Selectivity studies showed no interferences. Analysis of eight authentic samples collected from four subjects proved method feasibility. A simple, time-saving and green alternative for the analysis of DMT and harmala alkaloids in human urine samples was developed, optimized using design of experiments, fully validated and applied to authentic samples.

Stability Evaluation of DMT and Harmala Alkaloids in Ayahuasca Tea Samples.

Ayahuasca tea is a hallucinogenic beverage used for religious purposes in Brazil and many other countries that has therapeutic potential in the treatment of some mental health disorders. In the context of psychedelic research, quantification of the tea's main alkaloids prior to its administration in animal or human studies is essential. For this reason, this study aims to provide information regarding the stability of the main ayahuasca alkaloids (dimethyltryptamine, DMT; harmine, HRM; tetrahydroharmine, THH; harmaline, HRL) in three different conditions: (1) A year stored in a refrigerator either in plastic or glass containers, (2) seven days at 37 °C to reproduce usual mail transportation, and (3) after three freeze-thaw cycles. Samples were quantified after a dilute-and-shoot procedure using liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS). There was no significant degradation of DMT concentration over time in all tested conditions. Harmala alkaloids (THH, HRL, and HRM) showed important variations after long-term and high-temperature storages. Although DMT has proven to be stable in all studied conditions, the harmala alkaloids revealed intense degradation and even concentration increment. This may be caused by degradation, alkaloid inter-conversion, and leaching from tea precipitate material. Therefore, ayahuasca quantification before administration in controlled sets is mandatory.

DMH-CBD, a cannabidiol analog with reduced cytotoxicity, inhibits TNF production by targeting NF-kB activity dependent on A2A receptor.

Cannabidiol (CBD) is a natural compound with psychoactive therapeutic properties well described. Conversely, the immunological effects of CBD are still poorly explored. In this study, the potential anti-inflammatory effects and underlying mechanisms of CBD and its analog Dimethyl-Heptyl-Cannabidiol (DMH-CBD) were investigated using RAW 264.7 macrophages. CBD and DMH-CBD suppressed LPS-induced TNF production and NF-kB activity in a concentration-dependent manner. Both compounds reduced the NF-kB activity in a µM concentration range: CBD (IC50 = 15 µM) and DMH-CBD (IC50 = 38 µM). However, the concentrations of CBD that mediated NF-kB inhibition were similar to those that cause cytotoxicity (LC50 = 58 µM). Differently, DMH-CBD inhibited the NF-kB activation without cytotoxic effects at the same concentrations, although it provokes cytotoxicity at long-term exposure. The inhibitory action of the DMH-CBD on NF-kB activity was not related to the reduction in IkBα degradation or either p65 (NF-kB) translocation to the nucleus, although it decreased p38 MAP kinase phosphorylation. Additionally, 8-(3-Chlorostyryl) caffeine (CSC), an A2A antagonist, reversed the effect of DMH-CBD on NF-kB activity in a concentration-dependent manner. Collectively, our results demonstrated that CBD reduces NF-kB activity at concentrations intimately associated with those that cause cell death, whereas DMH-CBD decreases NF-kB activity at non-toxic concentrations in an A2A receptor dependent-manner.