Despite efforts to control synthetic cannabinoids, clandestine manufacturers continue to modify their structures to avoid legal consequences, creating an ever-changing analytical target for forensic laboratories (1). Forensic toxicology laboratories often lack the needed resources or do not have the capabilities to test for these compounds and metabolites, requiring specimens to be submitted to reference laboratories (2). Drug stability can be affected by long storage times, temperature and preservatives (3). Although these factors can be controlled, systematic research is necessary to identify their impacts on the stability of these new synthetic cannabinoids that are continually emerging. The purpose of this research is to assess the stability of 17 synthetic cannabinoids in human whole blood and 10 synthetic cannabinoid metabolites in human urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS) over thirty-five weeks. The analysis methods were validated in accordance to the Academy Standards Board (ASB) method validation guidelines for quantitative analysis and stability evaluation of the following analytes in blood: 4-cyano CUMYL-BUTINACA, ADB-PINACA, EMB-FUBINACA, JWH-250, MO-CHMINACA, 5-fluoro-3,5-ABPFUPPYCA, 5-fluoro ADB-PINACA, APP-PICA, CUMYL-THPINACA, PB-22, XLR11, 5-fluoro PY-PINACA, MDMB-FUBICA, MEP-CHMICA, NM2201, RCS-8, and UR144. The stability analysis in urine includes the following metabolites: 5-fluoro MDMB-PICA metabolite 7, 5-fluoro PB-22 3-carboxyindole, AB-FUBINACA metabolite 3, ADB-PINACA N-(4-hydroxypentyl), ADB-PINACA pentanoic acid, UR-144 Degradant N-pentanoic acid, PB-22 N-(5-hydroxypentyl), MDMB-FUBICA metabolite 3, UR-144 N-(5-hydroxypentyl), and JWH-250 N-pentanoic acid. Research samples were prepared by spiking with certified reference standards (Cayman Chemical, Ann Arbor, MI, USA) of each select synthetic cannabinoid in certified drug-free human whole blood (Boston Medical Center, Boston, MA, USA; Biological Specialty Corporation, Colmar, PA) and drug-free urine that was received as donations following the approved Institutional Review Board guidelines (Boston University School of Medicine, Boston, MA, USA). Blood samples were aliquoted into 6 mL BD Vacutainer Plastic Collection Tubes (Fisher Scientific, Waltham, MA, USA) and urine samples were stored in 15 mL Falcon Conical Centrifuge Tubes (Fisher Scientific, Waltham, MA, USA). Stability under room temperature (20ºC), refrigerator (4ºC), and freezer (-20ºC) at low and high concentrations were evaluated at select time points. A 5% solution of potassium oxalate and sodium fluoride or ethylenediaminetetraacetic acid (EDTA) was added to the preserved blood samples by the manufacturer prior to storage. The anticoagulant, potassium oxalate, was only added in solution to the preserved samples whereas none was added to the nonpreserved samples. Short-term urine samples were preserved with 1% of sodium fluoride prior to storage. Extraction of analytes was conducted using supported-liquid extraction (SLE) ISOLUTE 1 mL cartridges (Biotage, Charlotte, NC, USA) and reconstituted in 100 μL of 50:50 mixture of 0.1% formic acid in millipore deionized water and 0.1% formic acid in acetonitrile (Fisher Scientific, Waltham, MA, USA). Analysis was performed in triplicate using a reverse-phase C18 column (Waters XBridge C18 3.5 μM, 2.1 x 50 mm, Milford, MA, USA) on the Shimadzu Prominence Ultra-Fast Liquid Chromatography (UFLC, Kyoto, Japan) with SCIEX 4000 Q-Trap Electrospray Ionization Tandem Mass Spectrometry (ESI/MS/MS, Waltham, MA, USA) in positive ionization mode. The total run time was 8 minutes with a flow rate of 0.6 mL/min and injection volume of 10 μL. Linear calibration curves for each analyte with the exception of a quadratic regression for PB-22, all had acceptable R2 values > 0.99 using a weighting factor of 1/x. A linear dynamic range of 0.5 – 25 ng/mL was used for all analytes in blood except for NM2201 and APP-PICA with a limit of quantitation (LOQ) of 0.1 ng/mL and MO-CHMINACA with a working range of 0.5 – 15 ng/mL. A linear working range of 5 – 40 ng/mL was utilized for all metabolites in urine. No signs of carryover were observed. In general, analytes were considered stable if the average area ratio between the analyte and internal standard at the time point was within ± 20% of the average area ratio response at time point zero. In some cases, it was necessary to evaluate the complete picture of the stability data by reviewing analyte area, concentration, and overall stability data trend between timepoints at the low and high concentrations. In certain situations, an analyte was considered stable even if specific timepoints for a concentration were outside the ±20% range. For example, in cases where one concentration at a timepoint was within the ±20% range and the other concentration fell within ±30% range the analyte was considered stable overall. Long-term stability results revealed that all synthetic cannabinoids were stable at 21 to 35 weeks in frozen blood preserved with sodium fluoride except for APP-PICA. The preservatives are recommended to be added to blood to reduce the possibility of matrix inferences and minimize detrimental impacts on the stability of synthetic cannabinoids. Analytes experienced lower degradation in the order of samples that were kept frozen, refrigerated, and then at room temperature. Blood analytes that were stable up to 35 weeks in freezer generally had a core structure of a carbonyl substituent on a pyrazole or pyrrole with surrounding nonpolar groups; whereas compounds with two polar carbonyl functional groups present were found to experience degradation much earlier at 1 week or less in room temperature and refrigerator storage conditions. 5-fluoropentyl analogs, like XLR11 and 5-fluoro ADB-PINACA, in comparison to their counterpart analyte, UR144 and ADB-PINACA, were unstable at earlier time points under all storage conditions. Instability in the majority of the urine metabolites was not observed until after 9 weeks and was generally consistent across all storage conditions. The validated methods demonstrate a sensitive and reliable way to positively identify 17 different synthetic cannabinoids in human whole blood and 10 synthetic cannabinoid metabolites in urine for rapid time stability analysis at various storage conditions. The use of SLE improved sample preparation efficiency by decreasing the extraction time from 1 hour to 30 minutes compared to traditional extraction methods, such as solid-phase extraction (SPE) and liquid-liquid extraction (LLE). Further studies into additional matrices, such as oral fluid, longer storage times, and other emerging synthetic cannabinoid analytes would expand the scope of this research.

This thesis deals with the attitudes of students of two secondary schools to marihuana, alcohol use and their users. Attitudes to alcohol, marihuana and to their users are topics frequently discussed in connection with changing perception of these two substances. This trend can be best seen in adolescents during the period of late adolescence which is the period of secondary school studies. The main aim of the thesis is to find out which factors influence the attitudes of students to alcohol and drug use and if there are any differences concerning those factors between the observed schools. These factors come from theoretical considerations which are subsequently tested in research. As the research showed, the students' attitudes to marihuana and alcohol use, and also to alcohol users are primarily influenced by significant others, among which belong mainly peers and their parents or their own experience. While the individuals form their attitudes to alcohol primarily according to their parents, in case of marihuana, the individuals, who have experience with it, follow mainly their peers and their own experience. Those individuals also tend to be more positive towards other marihuana users. There have not been found out any significant differences concerning the attitudes to marihuana and alcohol...

"Benzo Fury" compounds and derivatives are enactogens similar to 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in various aspects. These compounds are similar in structure to MDMA and MDA, as well as, elicit similar effects such as elevated mood, euphoria and hallucinations. This similarity in effect increases the potential for abuse as MDMA has become less prevalent in some regions as the use of these new psychoactive substances (NPSs) has increased. The benzofury compounds are used as legal alternatives to MDMA because of their marketing as “not for human consumption”. With the relative ease in obtaining NPSs via the Internet, it is possible that these drugs may soon be prevalent in the United States. The project’s goal was to separate, detect, and quantitate the benzofury compounds and derivatives as well as MDA and MDMA in one method of analysis using ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS). The project also examined which method of sample preparation is more effective for these compounds. Six benzofury compounds were researched: 5-(2-aminopropyl)benzofuran) (5-APB), 6-(2-aminopropyl)benzofuran) (6-APB), 5-(2-aminopropyl)-2,3-dihydrobenzofuran (5-APDB), 6-(2-aminopropyl)-2,3-dihydrobenzofuran (6-APDB), 1-(benzofuran-5-yl)-N-methylpropan-2-amine (5-MAPB) and 1-(benzofuran-6-yl)-N-methylpropan-2-amine (6-MAPB) as well as MDMA and MDA. These drugs were analyzed in blood. A liquid-liquid extraction (LLE) method and solid phase extraction (SPE) method were examined to determine which would be better for the separation, detection and quantitation of the benzofury compounds. For the development of the overall method, accuracy, precision, calibration curve, carryover, limit of detection, limit of quantitation, analyte stability, and recovery were examined. The accuracy of the methods examined was greater than +/- 20%. For most analytes, the precision within-run and between-run did not exceed 20%, regardless of the sample preparation method used. A weighting of 1/x was applied to the calibration curve regardless of sample preparation method utilized. The carryover was less than 2% with the SPE method having less carryover (0.02% to 0.50%) than the LLE method (0.05% to 1.56%). The limit of quantitation was determined to be greater than 10 ng/mL. While this was unexpected, the limit of detection calculations determined that this was correct. Using the LLE method in combination with the UFLC-MS/MS method developed, the limit of detection was determined to be at least 9.98 ng/mL. Compared to the LLE method, the SPE limit of detection was lower and calculated to be 3.75 ng/mL. The percent recovery was examined for each of the analytes. It was determined that the SPE was capable of recovering 80% or more of the benzofury compounds and derivatives regardless of the concentration level. The LLE was not as successful in the recovering the benzofury compounds, the best recovery occurred at the 200 ng/mL level with only 65% or less recovered. Analyte stability exhibited a general decrease with variation prior to day 7 and then remains relatively stable until day 14. It was anticipated that the quantitation of the drugs might be complicated due to the similarity in structure between the isomers as well as the similarity of structure between all of the compounds. While this may still be the case, the difficult separation resulted in a re-evaluation and alterations to the UFLC-MS/MS method to correct for these issues. With the change in the UFLC-MS/MS method, further method optimization is required to achieve the appropriate accuracy and limit of quantitation. It was found that the best combination of sample preparation and detection of the benzofury compounds and derivatives is to use SPE followed by an UFLC-MS/MS method.

O diagnóstico duplo, ou comorbidade, é considerado prevalente tanto pela literatura do discurso médico-psiquiátrico, como na realidade clínica de serviços de saúde mental e de drogas. Considera-se que o campo da saúde está imerso em sistemas de crenças, influenciadas por aspectos culturais e sociais, inclusive sobre o que se considera sofrimento psíquico como sinônimo de diagnóstico psiquiátrico, que inserido em contexto mais amplo, se conectaria a sofrimentos compartilhados socialmente. O consumo de drogas estaria imerso nesse mesmo contexto histórico e social compartilhado. Este trabalho se caracterizou como pesquisa qualitativa, cuja abordagem conversou com autores que consideram a pesquisa qualitativa como um exercício de sensibilidade, cujo foco na experiência em primeira pessoa. O objetivo geral foi conhecer sentidos e significados do consumo de drogas a partir de narrativas de história de vida de pessoas descritas com duplo diagnóstico. O objetivos específicos foram conhecer descrições sobre a experiência de sofrimento psíquico ao longo da vida; conhecer descrições sobre a experiência de consumo de drogas ao longo da vida; e conhecer descrições sobre experiências em que o consumo de drogas esteja associado a sofrimentos psíquicos. Este estudo exploratório, foi realizado em um CAPS II e um CAPS AD, da rede de atenção psicossocial, de um município do interior paulista. Participaram dezoito adultos, homens e mulheres, selecionados de maneira a diversificar possibilidades de quadros clínicos e consumo de drogas, em padrão considerado problemático. A coleta de dados foi feita por meio de entrevista de história de vida temática e anotações em caderno de campo. A análise foi feita a partir do material emergente de trabalho de campo, da literatura sobre diagnóstico duplo e sobre sofrimento social. As entrevistas foram consideradas narrativas, sendo selecionadas dez para sua apresentação pormenorizada, cujos critérios de seleção foram exequibilidade e diversificação dos quadros clínicos, sendo distribuídos quanto ao gênero e serviços onde eram atendidos. Os resultados foram apresentados juntamente com a discussão, foi descrito o processo de construção do campo e do tema estudado e depois, foram descritas as narrativas. As narrativas dos participantes, na sua maioria homens e vindos de classes trabalhadoras urbanas, foram circunscritas pelo universo cultural e social onde se inseriam, mas também pelas possibilidades de escuta de demandas dos serviços da rede estudada. Esta se conformou em serviços que os descreviam de maneira excludente entre usuários de drogas e doentes mentais, estando comprometidas com perfis construídos nas diversas relações sociais e institucionais estabelecidas, considerando-se contextos e discursos mais amplos. Os sentidos e significados do consumo de drogas para os participantes foram construídos ao longo da vida, junto aos diversos eventos e situações vividos nas diversas relações sociais e culturais, em que o sofrimento psíquico, por vezes, fazia presente. O consumo de drogas, compreendido como dispositivo, permitia agir diante das situações de sofrimentos psíquicos e sociais, vividos singularmente, mas compartilhados socialmente. Considera-se importante refletir sobre as experiências vividas por pessoas em sofrimento psíquico e que fazem uso de drogas, considerando-os inseridos em um contexto mais amplo de relações sociais e institucionais, em que sofrimentos sociais estão presentes / Dual diagnosis, or comorbidity, is considered prevalent by literature of medical-psychiatric discourse, as much as, in mental health services and those for drug use treatment. It is considered that the health field is immerse in systems of beliefs influenced by cultural and social aspects, including what is considered psychological suffering as a synonym of psychiatric diagnosis, which is, in a broader context, connected with sufferings shared socially. Drug use is also immersed in the same historical and social shared context. This work is characterized as qualitative research, whose approach is related to authors that consider qualitative research as a sensibility exercise , focusing in first person experience. The main objective was to acknowledge senses and meanings of drug use from life history narratives of persons described as dually diagnosed. The specific objectives were to acknowledge descriptions of drug use along life course, descriptions of psychological suffering along life course and descriptions of drug use associated to psychological suffering. This exploratory study has been developed in one CAPS II and one CAPS AD, part of psychosocial attention network of an inner city of São Paulo. Eighteen adults participated, men and women, selected to permit diversification of clinical presentations of psychiatric diagnosis and drug use, in pattern od use considered problematic. Data collection has been thematic life histories interview and field notes. Analysis emerged from fieldwork, dual diagnosis\' literature and social suffering\'s literature. Interviews were considered narratives, and ten were presented with more details, selected by practicability, diversification of clinical presentations criteria, distributed among genders and services where they came from. Results were presented concomitantly with discussion, and were described the process of construction of the field and of the theme studied, then narratives were described and discussed. Participants narratives, mainly men from urban labourers class, were circumscribed by cultural and social universe where they came from, as well as the possibility of storytelling to be heard in the health services network studied. This network was also constructed by describing their patients exclusively as drug users or mentally ill, compromised with profiles constructed in many social and institutional relationships, considering broader context and discourses. Senses and meanings of drug use for the participants were constructed along life course, together with diverse events and situations lived in diverse social and cultural relations, and social and psychological sufferings could, sometimes, be present. Drug use, understood as a device, allowed the participant to act in situations of psychological sufferings that were experienced singularly, but socially shared. It is considered important reflect upon experiences lived by people in psychological suffering that also use drugs, considering them as part as a broader context of social and institutional relations, in which social sufferings may be present

RESUMO GONÇALVES, A.M. Cuidados diante do abuso e da dependência de drogas: desafio da prática do Programa Saúde da Família. 2002. 209fl. Tese. (Doutora-do em Enfermagem Psiquiátrica) – Escola de Enfermagem de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto. Este estudo tem como objeto a prática de cuidados relacionados ao abuso e à dependência de drogas no cotidiano de uma equipe do Programa Saúde da Família (PSF). Reconhecendo a gravidade dos problemas relaciona-dos ao abuso e à dependência de drogas e suas implicações para o indivíduo, a família e a comunidade, que refletem no processo de formação e na prática dos profissionais, tivemos como objetivo desenvolver com os membros de uma equipe do PSF uma ação-reflexão sobre essa prática, visando contribuir para a superação de problemas e contradições enfrentados pela equipe no cotidiano de trabalho. Realizamos uma investigação com abordagem qualitativa, na mo-dalidade de pesquisa-ação, fundamentada pelo materialismo histórico-dialético como parte intrínseca da visão de mundo vinculada à práxis. O cenário da pes-quisa foi a área geopolítica de atuação de uma equipe do Programa no municí-pio de Contagem, Estado de Minas Gerais, Brasil. A pesquisa de campo foi realizada no período de setembro de 2000 a julho de 2001 e teve como sujeitos os integrantes da equipe (um médico, uma enfermeira, uma auxiliar de enfer-magem e quatro agentes comunitários de saúde). Os resultados da pesquisa confirmaram a tese inicial sobre a importância do PSF para a prática de cuida-dos relacionados ao abuso e à dependência de drogas. Os resultados eviden-ciaram fragilidades e potencialidades técnicas (práticas e políticas) do Progra-ma para enfrentar esse desafio, além de apontar possibilidades de superação mediante investimento na capacitação da equipe e na criação de um projeto de ação comunitária. A pesquisa contribuiu com subsídio para formulação de polí-ticas de saúde, voltadas para o desafio de desenvolver cuidados relacionados ao abuso e à dependência de drogas na prática do PSF, bem como de outros serviços de saúde que venham valer-se dessa experiência para aplicá-la em condições semelhantes. / This study has as object the practice of cares related to the abuse and to the dependence of drugs in the daily of a team of the Program Health of the Family (PHF). Recognizing the gravity of the problems related to the abuse and to the dependence of drugs and their implications for the individual, the family and the community, that reflect in the formation process and in practice of the professionals, we had as objective develops with the members of a team of PHF an action-reflection on that practice, seeking to contribute to overcome the problems and contradictions faced by the team in the daily of work. We accom-plished an investigation with qualitative approach, in the research-action modal-ity, based by the materialism historical-dialectic as intrinsic part of the world vi-sion linked to the praxis. The scenery of the research was the area geopolitics of performance of a team of the Program in the municipal district of Contagem, State of Minas Gerais, Brazil. The field research was accomplished in the pe-riod from September of 2000 to July of 2001 and had as subjects the members of the team (a doctor, a nurse, a nursing auxiliary and four community agents of health). The results of the research confirmed the initial theses about the impor-tance of PHF for the practice of cares related to the abuse and to the depend-ence of drugs. The results evidenced fragilities and technical potentialities (practices and politics) of the Program to face that challenge, besides pointing possibilities to overcome that by investment in the training of the team and in the creation of a project of community action. The research contributed with subsidy to articulate politics of health, come back for the challenge from devel-oping cares related to the abuse and to the dependence of drugs in practice of the PHF, as well as of other services of health that come to be worth of that ex-perience to apply it in similar conditions.

A sociedade brasileira marcada pelo traço de desigualdade social é um dos cenários no qual a questão do uso de álcool e drogas coexiste com as condições e relações sociais, arranjos familiares e situações de estresse. Assim, a dependência de substância, as co-morbidades, modalidades de tratamento e recaídas durante o processo de reabilitação são fatores que devem ser contextualizados neste cenário. Em relação às políticas públicas a Organização Mundial da Saúde, no âmbito da saúde mental, prevê o tratamento dos portadores de sofrimento psíquico como seres sociais visando a reintegração destes sujeitos na comunidade; o Ministério da Saúde, na proposta do Sistema Único de Saúde por sua vez, preconiza a atenção integral aos usuários de álcool e outras drogas priorizando a atenção primária, a educação em saúde, garantia de atenção na comunidade e envolvimento das redes sociais. Os estudos recentes trazem à luz a importância da abordagem da rede e apoio social nesta área como possibilidade de repensar e propor novas perspectivas para o aprimoramento da pesquisa e da prática em saúde mental com foco nos indivíduos dependentes de substâncias psicoativas. Portanto o presente estudo diz respeito ao apoio social e uso de drogas considerando os sujeitos inseridos em seus contextos de relações e condições sociais. Logo o objetivo proposto foi o de analisar a diferença entre dependentes e não dependentes de substâncias quanto à percepção de apoio, utilizando um instrumento de medida de apoio social validado, e identificar possíveis especificidades da rede de apoio destes indivíduos. O referencial teórico metodológico adotado é o de apoio social na perspectiva do Buffering Model, e a Análise de Redes Sociais (ARS). A amostra dos sujeitos foi composta por indivíduos do Centro de Atenção Psicossocial álcool e drogas (CAPSad) e de uma Unidade Básica Distrital de Saúde (UBDS) do município de Ribeirão Preto totalizando 102 indivíduos sendo 50 dependentes e 52 não dependentes de substâncias. Foram observados os aspectos éticos da pesquisa envolvendo seres humanos a saber, aprovação do Comitê de Ética em Pesquisa, consentimento livre e esclarecido e observação do Código de Ética dos Profissionais de Enfermagem. Quanto aos resultados a amostra foi homogênea em relação às baixas condições sócio-econômicas e escore referente ao número de apoiadores. No entanto, em relação ao escore de satisfação com o apoio social os dependentes de substâncias mostraram-se menos satisfeitos com o apoio social disponível quando comparados com os não dependentes de drogas (diferença estatisticamente significante). Apesar disso, os indivíduos do CAPSad apresentaram redes mais diversificadas em sua composição enquanto os não dependentes de drogas apresentaram redes mais densas centradas, de um modo geral, no círculo familiar. Convém destacar que entre os não dependentes de substâncias, 27% consomem álcool e ou outras drogas num padrão que, embora não caracterize dependência, de acordo com os pontos de corte do Questionário para Triagem do Uso de Álcool, Tabaco e outras Substâncias (ASSIST) seriam beneficiados da intervenção breve. A discussão dos resultados deste estudo, certamente dará uma contribuição importante tanto para direcionar as práticas terapêuticas vigentes quanto para subsidiar futuras investigações da área. / The brazilian society is marked by social inequality and is one of the scenarios in which the issue of alcohol and drugs coexists with economic conditions, family and social relationships and the different stress situations. Thus, substance dependence, co-morbidities, treatment modalities and relapse are factors that must be contextualized in this scenario. Regarding the mental health, the World Health Organization directs that persons with mental disorders are treated as social beings with the goal of reintegration of these individuals in the community. The proposal of the Brazilian Unified Health System recommends full attention to the users of alcohol and other drugs with priority for primary care, health education, ensuring attention to community involvement and social networks. Recent studies highlight the importance of addressing the social network and support of drug addicts as a possibility to reflect and propose new research and practice in mental health for individuals addicted to drugs. Therefore, the present study concerns the social support and drug use considering the subjects included in their contexts of social relations and conditions. The objective was to analyze the difference between drug addicts and no drug addicts in perception and structure social network for these individuals. The theoretical framework adopted is social support in view \"Buffering Model\" and the Social Network Analysis (SNA). The sample of subjects consisted of 50 drug addicts and 52 no drug addicts of public health services in Ribeirão Preto. The project was approved by the Ethics Research Committee. Results: the sample was homogeneous with regard to low socio-economic score and the perceived number social support. The satisfaction with social support available of drug addicts was less when compared to no drug addicts (statistically significant). Nevertheless, the no drug addicts showed denser networks focused in the family and drug addicts have more diverse composition of networks. Among no drug addicts, 27% consume alcohol and other drugs in a pattern that according Alcohol, Smoking and Substance Involvement Screening Test (ASSIST) would benefit from brief intervention. The discussion of the results of this study will give an important contribution to direct the therapeutic practices and support future investigations of the area.

Introduction: Over recent decades, there has been an increase in the availability and use of Novel Psychoactive Substances (NPS) all over the world. They include several classes of chemicals that mimic the effects of illicit drugs and have been purposefully introduced into the market to circumvent or undermine the purpose of legal regulation. Currently, there is information lacking on the pharmacology of these substances; however, the increasing number of cases and outbreaks of intoxications/deaths is becoming a cause for deepening concern. Multi-disciplinary research in the fields of biology, chemistry, clinical medicine and web analysis is needed to develop responses against this tidal wave. Aim: The overall aim of this project is to gain insights into pharmacological, neurochemical and molecular properties of selected NPS to provide a reliable background needed for detection, assessment, and management of NPS-related harms. A range of approaches and methodologies was employed and a spectrum of different fields of knowledge has been engaged to gain some understanding into the complex multi-faceted phenomenon of NPS. Methods: Different substances have been selected as targets for the present project according to the clinical pattern of toxicity raised by their worldwide use and the lack of scientific knowledge available about them. The methods employed were: in vitro quantitative autoradiography (to evaluate the binding properties of the novel SCs BB-22, 5F-PB-22, 5F-AKB-48 and STS-135 at the cannabinoid receptor type 1 and N-methyl-D-aspartate receptor; and the binding properties of the synthetic stimulants 5-IT and 2-DPMP at the dopamine transporter in rat brain slices); in vitro Fast Scan Cyclic Voltammetry (to assess the effects of BB-22 on evoked dopamine efflux and dopamine re-uptake half-life in nucleus accumbens brain slices); in vivo microdialysis (to monitor dopamine release in terminal areas of the reward system after acute administration of the synthetic cannabinoids BB-22, 5F-PB-22, 5F-AKB-48 and STS-135; the dieting aid compound 2,4-DNP; the synthetic stimulants 2-DPMP and D2PM in freely moving animals); in silico molecular docking (to investigate the intermolecular interactions of the SCs BB-22, 5F-PB-22, 5F-AKB-48 and STS-135, and other referent compounds, with a homology model of the rodent cannabinoid receptor type 1 (CB1R) and the crystal structure of the human CB1R); and a web-based analysis approach (to analyse the information provided by a range of fora communities on 4,4'-DMAR use, additionally critical reviewing the available evidence-based literature on this topic). Results: Our in vitro quantitative autoradiography studies, confirmed that the index compounds BB-22, 5F-PB-22, 5F-AKB-48 and STS-135, behave as highly potent CB1R ligands able to compete with the radioligand [3H]CP-55,940 in cortical and striatal brain slices. On the other hand, all synthetic cannabinoids tested were unable to compete with the radioligand [3H]MK-801 in the same cerebral areas, rejecting the hypothesis of their potential binding to the N-methyl-D-aspartate receptor (NMDAR) at all concentrations investigated. Consistent with previous in vitro studies, 5-IT and 2-DPMP behaved as highly potent dopamine transporter (DAT) ligands able to compete with the radioligand [125 I]RTI-121 in a concentration-dependent way in the Caudate Putamen (CPu) and Nucleus Accumbens (NAc) brain slices. Notably, 2-DPMP was able to displace the radioligand in both cerebral regions, starting from lower concentrations compared to 5-IT. In vitro Fast Scan Cyclic Voltammetry findings demonstrated that local application of the synthetic cannabinoid BB-22 in brain slices, was unable to change evoked dopamine efflux and dopamine reuptake time-constant in the NAc shell at any doses tested. The results obtained would suggest the relative contributions of complex neuronal circuits, either within or outside the NAc, whose modulation would interfere with the interactions between BB-22 and dopaminergic neurons and represent critical pathways accounting for some of the rewarding properties of BB-22 exposure. In vivo microdialysis outcomes suggested that all SCs tested could increase dopamine release in the NAc shell at specific doses, while no changes in dopamine output were observed in other areas of the reward system, namely NAc core and medial prefrontal cortex (mPFCx) after BB-22 administration. These outcomes provided a circumstantial pre-clinical evidence for a greater putative abuse liability of SCs compared to the natural compound found in cannabis (Δ9‐THC). Furthermore, the acute treatment with 2,4-DNP did not cause any change in dopamine release in the NAc shell and CPu rejecting the hypothesis of psychoactivity of this substance at the dose tested. On the other hand, the synthetic stimulant 2-DPMP elicited a comparable increase of dopamine (DA) release in the NAc shell and CPu at the higher doses tested, while D2PM caused a selective increase of DA release in the NAc shell, providing a circumstantial preclinical evidence for a putative abuse liability of this compound at the highest dose assessed. The in silico molecular docking studies demonstrated that the SCs BB-22, 5F-PB-22, 5F-AKB-48 and STS-135 interact with CB1 receptor residues that, according to previous mutation and computational studies, are considered crucial for synthetic cannabinoid binding recognition. Additionally, they share some interacting residues with other aminoalkylindole derivatives (e.g. WIN-55,212-2). The web-based analysis focused on 4,4'-DMAR, suggested that fora members co-operate in exchanging an extensive body of knowledge about this drug, and the recurring topics of discussion include: routes of administration and dosages; desired and undesired effects; comparison and association with other drugs and medications; overall impression; provision of harm reduction advice. This approach has been useful to better understand some of the clinical and psychopharmacological issues pertaining to 4,4'-DMAR. Conclusions: Overall, these studies provided new pharmacological, neurochemical and molecular knowledge on a range of Novel Psychoactive Substances essential for identifying potential therapeutical approaches against their use/abuse. The novelty of this project lies in the adoption of a multi-disciplinary approach involving a range of methodologies from different areas of expertise (neurobiology, pharmacology, chemistry, netnography) all integrated to clarify some aspects of the index NPS, which were not yet available in the current literature. Additional studies are needed to better explain short and long-term effects of the index NPS, their abuse potential, and their interactions with other drugs of abuse.

Nenhuma medição é realizada com perfeição absoluta, uma vez que todos os valores encontrados são aproximações do valor real e todas as medidas, independente de sua finalidade ou qualidade, possuem uma incerteza. A incerteza de medição é um parâmetro associado ao resultado, que caracteriza a dispersão em torno dos seus valores. O conceito de incerteza de medição já é adotado em laboratórios de calibração e também muito aplicado na área de engenharia; no entanto em análises toxicológicas esta abordagem ainda é recente e há poucos relatos na literatura científica. Portanto, este trabalho teve como objetivo o estudo da incerteza de medição em análises toxicológicas confirmatórias de substâncias psicoativas - anfetaminas (anfetamina e metanfetamina), ácido 11-nor-Δ9-tetraidrocanabinol carboxílico (THC-COOH) e benzoilecgonina - em urina, detectados pela técnica de cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). A microextração em fase líquida (LPME) mostrou-se eficaz na determinação de THC-COOH, e após a completa validação, o método desenvolvido foi aplicado na quantificação de amostras de urina de referência provenientes do National Institute of Standards and Technology (NIST) dos Estados Unidos da América (SRM1507b - NIST). As principais contribuições para a incerteza do método foram a concentração do analito, a acurácia, seguidos da precisão e do volume de amostra. A incerteza combinada obtida foi equivalente a 8%. A LPME também apresentou-se eficiente para a extração das anfetaminas e a incerteza combinada obtida por este método foi 2,1%. No método de detecção de benzoilecgonina, a principal fonte de incerteza foi a acurácia do método e o resultado da incerteza combinada da análise de uma urina de referência (SRM1508a - NIST) foi 4,8%. Todos os valores de incerteza de medição encontrados em nosso estudo estão de acordo com as normas e referências internacionais e também são condizentes com os valores estipulados pela NIST nos laudos de análise das amostras de referência. / Neither measurement is made with absolute perfection, once all the values are approximations of the actual value and all measures, independent of its purpose or quality, have an uncertainty. Measurement uncertainty is a parameter associated with the result, which characterizes the dispersion around their values. The concept of measurement uncertainty is already used in calibration laboratories and also widely applied in engineering, however, in toxicological analysis, this approach is recent and there are few reports in the scientific literature. Therefore, this work aimed to study the measurement uncertainty in confirmatory toxicological analysis of psychoactive drugs - amphetamines (amphetamine and methamphetamine), acid 11-nor-Δ9-tetrahydrocannabinol carboxylic acid (THC - COOH) and benzoylecgonine - in urine detected by the technique of gas chromatography-mass spectrometry (GC- MS). The liquid phase microextraction (LPME) was effective in the determination of THC-COOH, and after complete validation, the method was applied to the quantification of urine samples of reference from the National Institute of Standards and Technology (NIST) of United States of America (SRM1507b - NIST). The main contributions to the uncertainty of the method were the analyte concentration, accuracy, followed by the precision and the sample volume. The combined uncertainty obtained was equivalent to 8%. The LPME also presented efficient for the extraction of amphetamine and combined uncertainty obtained by this method was 2.1%. In the method of detection of benzoylecgonine, the main source of uncertainty was the accuracy of the method and the result of the combined uncertainty of the analysis of a urine reference (SRM1508a - NIST) was 4.8%. All values of measurement uncertainty found in our study are in accordance with international standards and references and are also consistent with the values stipulated by certificate of analysis of NIST reference samples.

Nenhuma medição é realizada com perfeição absoluta, uma vez que todos os valores encontrados são aproximações do valor real e todas as medidas, independente de sua finalidade ou qualidade, possuem uma incerteza. A incerteza de medição é um parâmetro associado ao resultado, que caracteriza a dispersão em torno dos seus valores. O conceito de incerteza de medição já é adotado em laboratórios de calibração e também muito aplicado na área de engenharia; no entanto em análises toxicológicas esta abordagem ainda é recente e há poucos relatos na literatura científica. Portanto, este trabalho teve como objetivo o estudo da incerteza de medição em análises toxicológicas confirmatórias de substâncias psicoativas - anfetaminas (anfetamina e metanfetamina), ácido 11-nor-Δ9-tetraidrocanabinol carboxílico (THC-COOH) e benzoilecgonina - em urina, detectados pela técnica de cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). A microextração em fase líquida (LPME) mostrou-se eficaz na determinação de THC-COOH, e após a completa validação, o método desenvolvido foi aplicado na quantificação de amostras de urina de referência provenientes do National Institute of Standards and Technology (NIST) dos Estados Unidos da América (SRM1507b - NIST). As principais contribuições para a incerteza do método foram a concentração do analito, a acurácia, seguidos da precisão e do volume de amostra. A incerteza combinada obtida foi equivalente a 8%. A LPME também apresentou-se eficiente para a extração das anfetaminas e a incerteza combinada obtida por este método foi 2,1%. No método de detecção de benzoilecgonina, a principal fonte de incerteza foi a acurácia do método e o resultado da incerteza combinada da análise de uma urina de referência (SRM1508a - NIST) foi 4,8%. Todos os valores de incerteza de medição encontrados em nosso estudo estão de acordo com as normas e referências internacionais e também são condizentes com os valores estipulados pela NIST nos laudos de análise das amostras de referência. / Neither measurement is made with absolute perfection, once all the values are approximations of the actual value and all measures, independent of its purpose or quality, have an uncertainty. Measurement uncertainty is a parameter associated with the result, which characterizes the dispersion around their values. The concept of measurement uncertainty is already used in calibration laboratories and also widely applied in engineering, however, in toxicological analysis, this approach is recent and there are few reports in the scientific literature. Therefore, this work aimed to study the measurement uncertainty in confirmatory toxicological analysis of psychoactive drugs - amphetamines (amphetamine and methamphetamine), acid 11-nor-Δ9-tetrahydrocannabinol carboxylic acid (THC - COOH) and benzoylecgonine - in urine detected by the technique of gas chromatography-mass spectrometry (GC- MS). The liquid phase microextraction (LPME) was effective in the determination of THC-COOH, and after complete validation, the method was applied to the quantification of urine samples of reference from the National Institute of Standards and Technology (NIST) of United States of America (SRM1507b - NIST). The main contributions to the uncertainty of the method were the analyte concentration, accuracy, followed by the precision and the sample volume. The combined uncertainty obtained was equivalent to 8%. The LPME also presented efficient for the extraction of amphetamine and combined uncertainty obtained by this method was 2.1%. In the method of detection of benzoylecgonine, the main source of uncertainty was the accuracy of the method and the result of the combined uncertainty of the analysis of a urine reference (SRM1508a - NIST) was 4.8%. All values of measurement uncertainty found in our study are in accordance with international standards and references and are also consistent with the values stipulated by certificate of analysis of NIST reference samples.

Orientador: Fernando Bastos / Banca: Álvaro Lorencini Junior / Banca: Jair Lopes Junior / Banca: Sérgio Mello Arruda / Banca: Luciana Maria Lunardi Campos / Resumo: A atuação dos professores de Ciências e Biologia na prevenção educacional do abuso de substâncias psicoativas (SPA) recorre necessariamente a um processo de aquisição de saberes relacionados com psicofarmacologia e toxicologia, disciplinas atualmente não disponibilizadas na formação inicial desses docentes. Estuda-se o papel desenhado para o sistema educacional no que se refere às metas de prevenção ao abuso de drogas pela nova estrutura legislativa e pelos recentes posicionamentos dos tribunais nos casos que envolvem esta problemática orientando-o fortemente a ser tratado sob a ótica da educação onde um dos atores principais é o professor. Incumbidos dessa especial responsabilidade, de elevada complexidade, se despreparados, esses professores serão obstatos de realizá-la com eficiência e eficácia. As questões respondidas nesta tese fixaram-se em elucidar os aspectos da necessidade e de como capacitar professores daquelas disciplinas para atuarem dentro das escolas como profissionais da prevenção ao uso abusivo de substâncias psicoativas (SPA). Concebeu-se um corpo de conhecimentos comportando os saberes científico e da educação para a saúde, cuidando-se que os docentes devam contemplar os aspectos emocional e racional que a abordagem do tema exige. A pesquisa de abordagem qualitativa foi aplicada junto a duas turmas de licenciandos concluintes do Curso de Ciências Biológicas da Faculdade de Ciência da UNESP-Bauru, durante dois semestres, aferindo-se os conhecimentos e concepções pré-existentes e os apropriados após a implementação da capacitação. A capacitação incluiu conhecimentos dos aspectos legais aplicáveis ao setor educacional e de Direito Penal, Processual Penal, práticos e relacionados com questões polêmicas passíveis de confrontação pelos docentes, em sala de aula. Estudou-se... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The performing of teachers of science and biology in the educational prevention of psychoactive substances (SPA) abuse necessarily uses a process of acquiring knowledge related to psychopharmacology, toxicoloy, subjects not currently available in the initial formation of teachers. This study reveals the role designed for the educational system with regard to the goals of prevention of drug abuse by the new legislative structure and the placement of the courts in recent cases involving this issue, advising it strongly to be treated from the perspective of education where one of the main actors is the teacher. Taking in consideration that this is a task of special responsibility and high complexity, if unprepared, these teachers will be hindered to do so with efficiency and effectiveness. The questions answered in this thesis were set up to elucidate the necessity's aspects to teacher's formations to work as professionals within the schools to prevent the abuse of psychoactive substances (SPA). It was conceive a knowledge's frame including scientific knowledge and health education, taking care to that teachers contemplate the emotional and rational aspects required by the subject approaching. The search of qualitative approach was applied among two classes of ending degree students of Biological Sciences, College of Sciences, UNESP-Bauru, for two semesters, checking up their knowledge and pre-existing conceptions and acquired after the implementation of training. The enable proceeding included applicable knowledge of legal aspects for the education, Criminal Law, Criminal Proceeding, and practical issues related to controversial subject of possible confrontation by teachers in the classroom. Was studied the Federal Act Nº 11343 of 8/23/2006, the current drug law. The formation's effectiveness was evaluated along the lines investigated in five state high schools... (Complete abstract click electronic access below) / Doutor