<p> Illicit drug abuse is a widespread issue in the United States and worldwide. Many methods  seek to ease the analytical workload required to collect, analyze, and identify these drugs. Paper  spray-mass spectrometry (PS-MS) is one response to this analytical workload as it offers a rapid,  affordable, and simple means for drug identification by mass spectrometry. This work centers on  the use of pressure-sensitive adhesive (PSA) lined paper as a PS-MS substrate for drug recovery  and detection. The use of PSA paper as a sampling and analysis substrate has been previously  established but is expanded herein with new capabilities and applications. Chapter 2 introduces  the combination of color tests followed by PS-MS for presumptive and confirmatory drug  identification. Three color tests (cobalt thiocyanate, Simon, or Marquis) were performed on the  PSA paper with subsequent drug confirmation occurring by PS-MS. Chapter 3 examines the use  of PSA paper and PS-MS for the recovery and detection of fentanyl, fentanyl precursors, and  analogs from shipping-related surfaces and in the presence of high amounts of cutting agents. The  use of a cartridge that accommodates a full-sized PSA paper ticket was also explored for drug  detection. Chapter 4 assesses PS-MS with PSA paper on portable MS instrumentation. Analyte  recovery and carryover as well as instrument robustness were evaluated. The color test and PS-MS protocol examined in Chapter 2 was also successfully applied to a portable MS instrument.  Application of PS-MS to the portable system highlights the potential fieldability of the technique. </p>

The extremely high rates of intimate partner violence (IPV) experienced by Black women in community supervision programs (CSPs) who use drugs represents a major public health concern given the vast overrepresentation of Black women in the criminal legal system compared to non-Hispanic white women due to racialized drug laws and policies. National IPV surveillance data suggest that the rates of IPV in this population may be even higher among Black women who have sex with women and men (WSWM) in CSPs who use drugs. However, there remains a dearth of research that centers the experience of Black WSWM. Fear of experiencing police violence and experiences of racial and sexual discrimination pose additional challenges for Black WSMW in CSPs who use drugs to access both IPV and a broader range of services. No studies, to date, have examined typologies of IPV and its association to accessing IPV-related services among Black women with multiple intersecting minoritized identities including substance use, sexual behavior, and criminal-legal involvement. To address these gaps, this dissertation: 1) Identified typologies of IPV; 2) Examined how membership to latent classes is associated with use of core IPV services; and 3) Explored underlying mechanisms that may link IPV class, sexual behavior, and access to and utilization of IPV-related services. This dissertation study uses a sequential explanatory mixed methods approach with 1) secondary baseline survey data from Project EWORTH, a NIDA-funded HIV intervention study of 352 Black, drug-involved women mandated to CSPs and 2) primary qualitative follow-up data with participants from the same study to inform findings from the secondary data analysis. This dissertation found positive significant associations between having had both male and female sexual partners and more types and greater severity of IPV. Additionally, there was a significant, positive association between more types and greater severity of IPV and lifetime use of an order of protection. WSWM had a significantly higher odds of lifetime use of a DV shelter. Of interest, WSWM moderated the effect of people experiencing more severe violence accessing DV shelters. Qualitative interviews revealed unique forms of IPV such as feeling coerced to take a criminal charge for their partner and spiritual abuse, both of which are not captured with standard IPV measures or discussed broadly in IPV literature. Additionally, CSP staff served as an important link to services among these women. Overall, these results suggest that more inclusive IPV screening, referral to service, and actual services, as well as providing training for service providers that consider the effects of multiple, marginalized identities has on experience of IPV, and access to and use of services among Black women in the criminal legal system.

The role of biological markers (biomarkers) in public health, pediatric medicine and clinical toxicology cannot be understated. Clinically validated biomarkers used in newborn screening (NBS) serve to detect individuals at risk for a disease in the population, pre-symptomatically diagnose affected neonates early in life and/or accurately predict disease progression and treatment responses to therapy. However, there is urgent need for the discovery of more specific biomarkers that can improve screening accuracy in a high throughput, cost-effective yet ethical manner. The major objectives of this thesis were to develop innovative nontargeted metabolite profiling methodologies based on multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS) for early detection of treatable genetic diseases, as well as comprehensive surveillance of drugs of abuse (DoA) in high risk subjects. Chapter II introduces a multiplexed MSI-CE-MS strategy for confirmatory testing of known biomarkers for various inborn errors of metabolism from a dried blood spot (DBS) that was rigorously validated using proficiency test specimens from Centres for Disease Control and Prevention (CDC) and authentic neonatal samples from Newborn Screening Ontario (NSO) with quality assurance. Additionally, MSI-CE-MS together with temporal signal pattern recognition revealed for the first time a novel class of pathognomonic marker elevated in galactosemia, namely N-galactated amino acids. Chapter III describes an untargeted metabolomic study to discover biomarkers of cystic fibrosis (CF) to reduce the high false positive rate and widespread carrier identification by current two-tiered NBS algorithms that rely on genetic testing. A panel of metabolites from retrospective DBS specimens, including several amino acids, ophthalmic acid and an unknown peptide, allowed for differentiation of asymptomatic CF neonates from screen-positive yet unaffected carriers and transient hypertrypsinogenemic cases. Chapter IV develops and validates a high throughput MSI-CE-MS assay for rapid screening for DoA and their metabolites in human urine with improved specificity and broad spectrum coverage as compared to classic targeted immunoassays. This method can also applied to ensure compliance, drug efficacy and patient safety while detecting for potential substitution or adulteration when using high resolution MS/MS. In summary, this thesis contributes an innovative methodology and data workflow for biomarker discovery for improved neonatal screening of rare genetic diseases in the population, which was also applied for more effective drug surveillance strategies in public health given the alarming worldwide opioid crisis. / Thesis / Doctor of Philosophy (PhD)

No / A novel dispersive system operating at 1064-nm excitation and coupled with transfer electron InGaAs photocathode and electron bombardment CCD technology has been evaluated for the analysis of drugs of abuse and explosives. By employing near-IR excitation at 1064-nm excitation wavelength has resulted in a significant damping of the fluorescence emission compared to 785-nm wavelength excitation. Spectra of street samples of drugs of abuse and plastic explosives, which usually fluoresce with 785-nm excitation, are readily obtained in situ within seconds through plastic packaging and glass containers using highly innovative detector architecture based upon a transfer electron (TE) photocathode and electron bombarded gain (EB) technology that allowed the detection of NIR radiation at 1064nm without fluorescence interference. This dispersive near-IR Raman system has the potential to be an integral part in the armoury of the forensic analyst as a non-destructive tool for the in-situ analysis of drugs of abuse and explosives. Copyright (c) 2015 John Wiley & Sons, Ltd.

Le système nerveux central est un organe privilégié et protégé, notamment grâce à l’existence des barrières histologiques entre le sang et les tissus nerveux. La barrière-hémato encéphalique (BHE) et la barrière hémato-rétinienne (BHR) séparent respectivement le parenchyme cérébral et la rétine des composés contenus dans l’espace vasculaire, grâce à l’expression de jonctions serrées et de transporteurs membranaires permettant une régulation spécifique des échanges entre le sang et le parenchyme nerveux. Ce travail a porté sur l’étude d’un nouveau transporteur de cations organiques mis en évidence fonctionnellement à la BHE de la souris. Ce transporteur appartenant très probablement à la superfamille des solute carrier (SLC), fonctionne comme un antiport proton. Actuellement, sa présence ne peut être démontrée que de façon fonctionnelle car son identité moléculaire est encore inconnue. Cet antiport proton constitue un nouvel acteur de la perméabilité cérébrale et ouvre une nouvelle voie d’accès au cerveau. Nous nous sommes tout d’abord attachés à approfondir les connaissances fonctionnelles de ce transporteur en étudiant de nouveaux substrats et tissus d’expression. Le transport cérébral de psychotropes a été étudié in vivo par la technique de perfusion carotidienne in situ chez la souris et in vitro grâce à une lignée de cellules endothéliales cérébrales humaines immortalisées (hCMEC/D3). Nous avons démontré que la haute perméabilité cérébrale de la cocaïne fait intervenir à la fois une diffusion passive et surtout une diffusion médiée par un antiport proton. La vitesse d’entrée des substances d’abus dans le cerveau est associée à un plus fort risque d’addiction et fait de ce transporteur un nouvel acteur critique de la régulation du passage cérébral. En effet, d’autres substances comme la nicotine et certaines amphétamines comme le MDPV et l'ecstasy sont également des substrats de cet antiport. Ce transporteur apparaît comme une cible pharmacologique potentielle dans la prise en charge de toxicomanies. Malgré la diversité chimique et pharmacologique d’interactions des composés avec cet antiport, les concentrations nécessaires pour l’inhiber dépassent celles retrouvées dans le sang. Pour aider l’identification d’inhibiteurs sélectifs et efficaces nous avons développé un modèle pharmacophorique d’inhibiteurs du transporteur à partir de données générées in vitro et de l’approche FLAPpharm. Ce modèle semble prédictif de nouveaux composés pouvant constituer de meilleurs inhibiteurs de ce transporteur. L’étude des échanges in vivo au niveau du tissu nerveux nous a menés à étudier l’impact de transporteurs ABC et de l’antiport-proton au niveau cérébral et rétinien à l’aide de substances spécifiques ou de substrats mixtes comme le vérapamil. L’antiport proton est fonctionnel au niveau de la BHR et transporte notamment la clonidine, le DPH et le vérapamil. Cependant, dans le cas d’un substrat mixte P-gp et SLC (ex : vérapamil), ce transport d’influx n’est visible à la BHE que lorsque la P-gp est neutralisée. Au contraire, à la BHR l’influx lié à cet SLC est visible naturellement. L’impact de la P-gp à la BHR étant 6.3-fois plus faible ce processus est probablement moins masqué. Cette étude illustre la difficulté actuelle de prédire l’impact fonctionnel d’un transporteur pour des substrats multi-spécifiques et l’existence d’une priorisation du transport. Enfin, nous avons essayé d’identifier l’antiport proton au niveau moléculaire par une méthode de photo-activation à l’aide d’un composé adapté. Cette méthode s’est avérée efficace pour fixer une molécule sur le transporteur, permettant par la suite de l’isoler plus facilement. En conclusion, ce travail a permis de mettre en évidence l’importance de l’antiport proton dans la distribution cérébrale de psychotropes et d’ouvrir de nouvelles perspectives dans l’addiction et la compréhension du transport de substrats multi-spécifiques. / The central nervous system is a privilege organ protected by histological barriers between the blood and the nervous tissue. The blood-brain barrier (BBB) and the blood-retinal barrier (BRB) separate cerebral parenchyma and retina from the circulating blood and both express tight junctions and membrane transporters, allowing a precise regulation of the exchanges between the blood and nervous tissues. We studied a new cationic transporter functionally evidenced at the mouse BBB. This molecularly unknown transporter belong to the solute carrier super family (SLC) and is a proton antiporter. It could constitute a new actor in the cerebral permeability and may be a new brain access pathway. First, we worked on the functional identification studying new substrates and new localization. Psychotropic brain transport was studied in vivo by brain in situ perfusion on mouse and in vitro with human immortalized endothelial cells (hCMEC/D3). We showed that cocaine brain entry depends on passive diffusion but also mainly on a proton antiporter. Brain entry rate of drugs of abuse is associated with modulation of addiction liability, making this transporter a new component of brain entry of cocaine, and also nicotine and some amphetamines such as ecstasy and MDPV. This proton antiporter appears to be a new potential target in addiction. Various chemical entities interact with this transporter; however concentrations used to inhibit the transporter are much higher than the one possibly found in the blood. In order to help find or design new selective and potent inhibitors, we developed a pharmacophore model of the proton antiporter inhibitors using in vitro data and the FLAPpharm approach. The model predicts well new possible inhibitors of this transporter. We also studied the impact of the ABC transporters and the proton antiporter at the BBB and the BRB using specific or multi-specific substrates such as verapamil. The proton antiporter is functionally expressed at the BRB and transports clonidine, DPH and verapamil. However, for the multi-specific (P-gp and SLC) compound verapamil, influx transport by the proton antiporter is visible at the BBB only when P-gp efflux is neutralized. On the contrary, at the BRB, the proton antiporter influx is always visible. This is certainly due to the lower impact (by 6.3 fold) of P-gp at the BRB compared to the BBB. These results show the difficulty to predict the functional impact of a transporter for multi-specific compounds and a probable transport prioritization. Finally we worked on the molecular identification of the proton antiporter using a photolabeling method. This work evidenced the importance of the proton antiporter in the brain distribution of psychotropic and drugs of abuse and opened toward new perspectives in addiction and transport comprehension.

Le système nerveux central est un organe privilégié et protégé, notamment grâce à l’existence des barrières histologiques entre le sang et les tissus nerveux. La barrière-hémato encéphalique (BHE) et la barrière hémato-rétinienne (BHR) séparent respectivement le parenchyme cérébral et la rétine des composés contenus dans l’espace vasculaire, grâce à l’expression de jonctions serrées et de transporteurs membranaires permettant une régulation spécifique des échanges entre le sang et le parenchyme nerveux. Ce travail a porté sur l’étude d’un nouveau transporteur de cations organiques mis en évidence fonctionnellement à la BHE de la souris. Ce transporteur appartenant très probablement à la superfamille des solute carrier (SLC), fonctionne comme un antiport proton. Actuellement, sa présence ne peut être démontrée que de façon fonctionnelle car son identité moléculaire est encore inconnue. Cet antiport proton constitue un nouvel acteur de la perméabilité cérébrale et ouvre une nouvelle voie d’accès au cerveau. Nous nous sommes tout d’abord attachés à approfondir les connaissances fonctionnelles de ce transporteur en étudiant de nouveaux substrats et tissus d’expression. Le transport cérébral de psychotropes a été étudié in vivo par la technique de perfusion carotidienne in situ chez la souris et in vitro grâce à une lignée de cellules endothéliales cérébrales humaines immortalisées (hCMEC/D3). Nous avons démontré que la haute perméabilité cérébrale de la cocaïne fait intervenir à la fois une diffusion passive et surtout une diffusion médiée par un antiport proton. La vitesse d’entrée des substances d’abus dans le cerveau est associée à un plus fort risque d’addiction et fait de ce transporteur un nouvel acteur critique de la régulation du passage cérébral. En effet, d’autres substances comme la nicotine et certaines amphétamines comme le MDPV et l'ecstasy sont également des substrats de cet antiport. Ce transporteur apparaît comme une cible pharmacologique potentielle dans la prise en charge de toxicomanies. Malgré la diversité chimique et pharmacologique d’interactions des composés avec cet antiport, les concentrations nécessaires pour l’inhiber dépassent celles retrouvées dans le sang. Pour aider l’identification d’inhibiteurs sélectifs et efficaces nous avons développé un modèle pharmacophorique d’inhibiteurs du transporteur à partir de données générées in vitro et de l’approche FLAPpharm. Ce modèle semble prédictif de nouveaux composés pouvant constituer de meilleurs inhibiteurs de ce transporteur. L’étude des échanges in vivo au niveau du tissu nerveux nous a menés à étudier l’impact de transporteurs ABC et de l’antiport-proton au niveau cérébral et rétinien à l’aide de substances spécifiques ou de substrats mixtes comme le vérapamil. L’antiport proton est fonctionnel au niveau de la BHR et transporte notamment la clonidine, le DPH et le vérapamil. Cependant, dans le cas d’un substrat mixte P-gp et SLC (ex : vérapamil), ce transport d’influx n’est visible à la BHE que lorsque la P-gp est neutralisée. Au contraire, à la BHR l’influx lié à cet SLC est visible naturellement. L’impact de la P-gp à la BHR étant 6.3-fois plus faible ce processus est probablement moins masqué. Cette étude illustre la difficulté actuelle de prédire l’impact fonctionnel d’un transporteur pour des substrats multi-spécifiques et l’existence d’une priorisation du transport. Enfin, nous avons essayé d’identifier l’antiport proton au niveau moléculaire par une méthode de photo-activation à l’aide d’un composé adapté. Cette méthode s’est avérée efficace pour fixer une molécule sur le transporteur, permettant par la suite de l’isoler plus facilement. En conclusion, ce travail a permis de mettre en évidence l’importance de l’antiport proton dans la distribution cérébrale de psychotropes et d’ouvrir de nouvelles perspectives dans l’addiction et la compréhension du transport de substrats multi-spécifiques. / The central nervous system is a privilege organ protected by histological barriers between the blood and the nervous tissue. The blood-brain barrier (BBB) and the blood-retinal barrier (BRB) separate cerebral parenchyma and retina from the circulating blood and both express tight junctions and membrane transporters, allowing a precise regulation of the exchanges between the blood and nervous tissues. We studied a new cationic transporter functionally evidenced at the mouse BBB. This molecularly unknown transporter belong to the solute carrier super family (SLC) and is a proton antiporter. It could constitute a new actor in the cerebral permeability and may be a new brain access pathway. First, we worked on the functional identification studying new substrates and new localization. Psychotropic brain transport was studied in vivo by brain in situ perfusion on mouse and in vitro with human immortalized endothelial cells (hCMEC/D3). We showed that cocaine brain entry depends on passive diffusion but also mainly on a proton antiporter. Brain entry rate of drugs of abuse is associated with modulation of addiction liability, making this transporter a new component of brain entry of cocaine, and also nicotine and some amphetamines such as ecstasy and MDPV. This proton antiporter appears to be a new potential target in addiction. Various chemical entities interact with this transporter; however concentrations used to inhibit the transporter are much higher than the one possibly found in the blood. In order to help find or design new selective and potent inhibitors, we developed a pharmacophore model of the proton antiporter inhibitors using in vitro data and the FLAPpharm approach. The model predicts well new possible inhibitors of this transporter. We also studied the impact of the ABC transporters and the proton antiporter at the BBB and the BRB using specific or multi-specific substrates such as verapamil. The proton antiporter is functionally expressed at the BRB and transports clonidine, DPH and verapamil. However, for the multi-specific (P-gp and SLC) compound verapamil, influx transport by the proton antiporter is visible at the BBB only when P-gp efflux is neutralized. On the contrary, at the BRB, the proton antiporter influx is always visible. This is certainly due to the lower impact (by 6.3 fold) of P-gp at the BRB compared to the BBB. These results show the difficulty to predict the functional impact of a transporter for multi-specific compounds and a probable transport prioritization. Finally we worked on the molecular identification of the proton antiporter using a photolabeling method. This work evidenced the importance of the proton antiporter in the brain distribution of psychotropic and drugs of abuse and opened toward new perspectives in addiction and transport comprehension.

Esta tese apresenta os estudos e esforços visando ao desenvolvimento de sensores químicos para aplicações diversas na área forense. Foram desenvolvidos métodos eletroanalíticos para detecção e quantificação de alguns compostos comumente encontrados na adulteração de amostras de drogas de abuso (procaína, fenacetina, aminopirina, paracetamol, levamisol), além da cocaína e estudos fundamentais sobre o comportamento eletroquímico desses compostos. Empregaram-se também métodos eletroquímicos para quantificação de compostos tóxicos e perigosos como explosivos (ácido pícrico) e melamina por exemplo. Os trabalhos utilizando sensores eletroquímicos contemplam modificações eletroquímicas das superfícies eletródicas, utilização de sensores com polímeros molecularmentes impressos (MIP) e eletrodos descartáveis em papel utilizando diferentes técnicas voltamétricas e amperométricas, eletrodo disco rotatório (EDR) e microbalança de cristal de quartzo. Além da fabricação de dispositivos analíticos descartáveis em papel empregando detecção eletroquímica utilizou-se também a detecção colorimétrica para quantificação de alguns dos principais adulterantes de amostras de apreensão de cocaína, como procaína e fenacetina, bem como análises e discriminações de compostos explosivos (peroxi e nitro compostos) nessas plataformas portáteis e de baixo custo. Os métodos foram sempre desenvolvidos visando característicos como: facilidade, praticidade, baixo custo e portabilidade para análises diretamente no local de medida com mínima infraestrutura laboratorial. Por fim, são apresentados alguns estudos realizados durante estágio de pesquisa no exterior (Universidade da Califórnia - San Diego (UCSD)) na área de Wearable Sensors, em que foram desenvolvidos métodos para análises de micronutrientes no suor (zinco) e um metabólito (ácido úrico) na saliva usando sensores aplicados diretamente no corpo humano. / This thesis shows studies and efforts to the development of chemical sensors for different applications in the forensic field. Electroanalytical methods were developed for detection and quantification of some compounds (procaine, phenacetin, aminopyrine, acetaminophen, levamisole) commonly found in the drug of abuse adulteration process and cocaine, as well as, fundamental studies about the electrochemical behavior of these compounds. It was also employed electrochemical methods for quantification of hazardous compounds such as explosives (picric acid) and melamine. Analytical methods with electrochemical sensors included electrochemical modification of electrodic surfaces, molecularly imprinted polymers (MIP), and paper disposable electrochemical devices using different voltammetric and amperometric techniques, rotating disc electrode (RDE) and quartz crystal microbalance. In addition to the fabrication of paper disposable analytical devices with electrochemical detection, it was also used the colorimetric detection to quantify some of the major adulterants in cocaine seizure samples, such as procaine and phenacetin, as well as analysis and discrimination of explosive compounds (peroxy and nitro explosives) in these low cost portable platforms. All proposed methods were always developed aming at theses characteristics: ease, convenience, low cost and portability for analysis directly at the measurement site with minimal laboratory infrastructure. Finally, we presented some studies conducted during research internship abroad (University of California - San Diego (UCSD)) in the area of Wearable Sensors, which have been developed methods for micronutrient analysis in sweat (Zn) and a metabolite (Uric Acid) in saliva using sensors applied directly to the human body

Le système nerveux central est un organe privilégié et protégé, notamment grâce à l’existence des barrières histologiques entre le sang et les tissus nerveux. La barrière-hémato encéphalique (BHE) et la barrière hémato-rétinienne (BHR) séparent respectivement le parenchyme cérébral et la rétine des composés contenus dans l’espace vasculaire, grâce à l’expression de jonctions serrées et de transporteurs membranaires permettant une régulation spécifique des échanges entre le sang et le parenchyme nerveux. Ce travail a porté sur l’étude d’un nouveau transporteur de cations organiques mis en évidence fonctionnellement à la BHE de la souris. Ce transporteur appartenant très probablement à la superfamille des solute carrier (SLC), fonctionne comme un antiport proton. Actuellement, sa présence ne peut être démontrée que de façon fonctionnelle car son identité moléculaire est encore inconnue. Cet antiport proton constitue un nouvel acteur de la perméabilité cérébrale et ouvre une nouvelle voie d’accès au cerveau. Nous nous sommes tout d’abord attachés à approfondir les connaissances fonctionnelles de ce transporteur en étudiant de nouveaux substrats et tissus d’expression. Le transport cérébral de psychotropes a été étudié in vivo par la technique de perfusion carotidienne in situ chez la souris et in vitro grâce à une lignée de cellules endothéliales cérébrales humaines immortalisées (hCMEC/D3). Nous avons démontré que la haute perméabilité cérébrale de la cocaïne fait intervenir à la fois une diffusion passive et surtout une diffusion médiée par un antiport proton. La vitesse d’entrée des substances d’abus dans le cerveau est associée à un plus fort risque d’addiction et fait de ce transporteur un nouvel acteur critique de la régulation du passage cérébral. En effet, d’autres substances comme la nicotine et certaines amphétamines comme le MDPV et l'ecstasy sont également des substrats de cet antiport. Ce transporteur apparaît comme une cible pharmacologique potentielle dans la prise en charge de toxicomanies. Malgré la diversité chimique et pharmacologique d’interactions des composés avec cet antiport, les concentrations nécessaires pour l’inhiber dépassent celles retrouvées dans le sang. Pour aider l’identification d’inhibiteurs sélectifs et efficaces nous avons développé un modèle pharmacophorique d’inhibiteurs du transporteur à partir de données générées in vitro et de l’approche FLAPpharm. Ce modèle semble prédictif de nouveaux composés pouvant constituer de meilleurs inhibiteurs de ce transporteur. L’étude des échanges in vivo au niveau du tissu nerveux nous a menés à étudier l’impact de transporteurs ABC et de l’antiport-proton au niveau cérébral et rétinien à l’aide de substances spécifiques ou de substrats mixtes comme le vérapamil. L’antiport proton est fonctionnel au niveau de la BHR et transporte notamment la clonidine, le DPH et le vérapamil. Cependant, dans le cas d’un substrat mixte P-gp et SLC (ex : vérapamil), ce transport d’influx n’est visible à la BHE que lorsque la P-gp est neutralisée. Au contraire, à la BHR l’influx lié à cet SLC est visible naturellement. L’impact de la P-gp à la BHR étant 6.3-fois plus faible ce processus est probablement moins masqué. Cette étude illustre la difficulté actuelle de prédire l’impact fonctionnel d’un transporteur pour des substrats multi-spécifiques et l’existence d’une priorisation du transport. Enfin, nous avons essayé d’identifier l’antiport proton au niveau moléculaire par une méthode de photo-activation à l’aide d’un composé adapté. Cette méthode s’est avérée efficace pour fixer une molécule sur le transporteur, permettant par la suite de l’isoler plus facilement. En conclusion, ce travail a permis de mettre en évidence l’importance de l’antiport proton dans la distribution cérébrale de psychotropes et d’ouvrir de nouvelles perspectives dans l’addiction et la compréhension du transport de substrats multi-spécifiques. / The central nervous system is a privilege organ protected by histological barriers between the blood and the nervous tissue. The blood-brain barrier (BBB) and the blood-retinal barrier (BRB) separate cerebral parenchyma and retina from the circulating blood and both express tight junctions and membrane transporters, allowing a precise regulation of the exchanges between the blood and nervous tissues. We studied a new cationic transporter functionally evidenced at the mouse BBB. This molecularly unknown transporter belong to the solute carrier super family (SLC) and is a proton antiporter. It could constitute a new actor in the cerebral permeability and may be a new brain access pathway. First, we worked on the functional identification studying new substrates and new localization. Psychotropic brain transport was studied in vivo by brain in situ perfusion on mouse and in vitro with human immortalized endothelial cells (hCMEC/D3). We showed that cocaine brain entry depends on passive diffusion but also mainly on a proton antiporter. Brain entry rate of drugs of abuse is associated with modulation of addiction liability, making this transporter a new component of brain entry of cocaine, and also nicotine and some amphetamines such as ecstasy and MDPV. This proton antiporter appears to be a new potential target in addiction. Various chemical entities interact with this transporter; however concentrations used to inhibit the transporter are much higher than the one possibly found in the blood. In order to help find or design new selective and potent inhibitors, we developed a pharmacophore model of the proton antiporter inhibitors using in vitro data and the FLAPpharm approach. The model predicts well new possible inhibitors of this transporter. We also studied the impact of the ABC transporters and the proton antiporter at the BBB and the BRB using specific or multi-specific substrates such as verapamil. The proton antiporter is functionally expressed at the BRB and transports clonidine, DPH and verapamil. However, for the multi-specific (P-gp and SLC) compound verapamil, influx transport by the proton antiporter is visible at the BBB only when P-gp efflux is neutralized. On the contrary, at the BRB, the proton antiporter influx is always visible. This is certainly due to the lower impact (by 6.3 fold) of P-gp at the BRB compared to the BBB. These results show the difficulty to predict the functional impact of a transporter for multi-specific compounds and a probable transport prioritization. Finally we worked on the molecular identification of the proton antiporter using a photolabeling method. This work evidenced the importance of the proton antiporter in the brain distribution of psychotropic and drugs of abuse and opened toward new perspectives in addiction and transport comprehension.

Submitted by admin tede (tede@pucgoias.edu.br) on 2016-11-22T12:24:00Z No. of bitstreams: 1 FLAVIA MARIA PEREIRA SIMÃO MIRANDA.pdf: 981229 bytes, checksum: 3b1aaa70f2d5eeda217acc6312f17009 (MD5) / Made available in DSpace on 2016-11-22T12:24:00Z (GMT). No. of bitstreams: 1 FLAVIA MARIA PEREIRA SIMÃO MIRANDA.pdf: 981229 bytes, checksum: 3b1aaa70f2d5eeda217acc6312f17009 (MD5) Previous issue date: 2016-06-29 / It sought, through a historical-legal, doctrinal study, analyze the problem of drugs and the growing crime resulting from the use, abuse or dependence on narcotic substances, which requires scholars to develop new alternatives to the traditional model of law criminal and aims to implement a more effective fundamental rights, in particular the right to life, which stems from the principle of human dignity (one of the foundations of the Federative Republic of Brazil, pursuant to art. 1 of the Federal Constitution), the right to physical, mental and social integrity and the right to health. It adopts the method of hypothetical-deductive approach, with literature review. It presents an overview of the Brazilian prison system, demonstrating the failure of deprivation of liberty and its ineffectiveness because, despite the increase in prisoners, it did not have the power to reduce crime. It is observed from the study that the Institute of Therapeutic Justice is presented as a viable and effective solution to the growing of the crime problems related to drug use, as it is an excellent strategy for coping with addiction binomial and/or drug use and crime. It is being implemented throughout the Brazilian territory, as it has high levels of efficiency. He emerged in Brazil following the American model of Drug Courts, but with its own peculiarities, which will be demonstrated in this study. It aims to analyze the mechanisms used by the program, its usefulness, scope, applicability in the Brazilian legal system and the results obtained from its action within the State of Goiás Judiciary. Specifically, one of the foundations for the research was a shortage of about stuff theme, therefore, is intended to guide the professionals of law and spread the importance of the institute, aiming at greater implementation of the program and also effective participation of society. / Buscou-se, por meio de um estudo histórico-legislativo, doutrinário, analisar a problemática das drogas e a crescente criminalidade decorrente do uso, abuso ou dependência de substâncias entorpecentes, a qual exige dos estudiosos o desenvolvimento de novas alternativas ao modelo tradicional do Direito Penal, bem como almeja implementar uma maior efetividade aos direitos fundamentais, em específico o direito à vida, do qual decorre o princípio da dignidade da pessoa humana (um dos fundamentos da República Federativa do Brasil, nos termos do art. 1º, da Constituição Federal), o direito à integridade física, psíquica e social e o direito à saúde. Adota o método de abordagem hipotético-dedutivo, com análise bibliográfica. Apresenta o panorama do sistema prisional brasileiro, demonstrando a falência da pena privativa de liberdade e a sua ineficácia, pois, apesar do aumento de presos, isto não teve o condão de diminuir a criminalidade. Observa-se a partir do estudo que o instituto da Justiça Terapêutica se apresenta como solução viável e eficaz para a problemática crescente do delito relacionado ao uso de drogas, tratando-se de uma excelente estratégia para o enfrentamento do binômio dependência química e/ou uso de drogas e criminalidade. Está sendo implementada em todo o território brasileiro, pois apresenta altos níveis de eficácia. Despontou no Brasil seguindo o modelo norteamericano das Drug Courts, mas, com suas peculiaridades, as quais serão demonstradas no presente estudo. Objetiva analisar os mecanismos utilizados pelo programa, sua utilidade, abrangência, aplicabilidade no ordenamento jurídico brasileiro e os resultados obtidos a partir da sua intervenção dentro do Poder Judiciário do Estado de Goiás. Especificamente, um dos fundamentos para a pesquisa foi a escassez de material acerca do tema, pois, pretende orientar os profissionais do Direito e difundir a importância do instituto, visando a maior implementação do programa e também participação efetiva da sociedade.

O objetivo deste estudo foi compreender os conceitos e estratégias que norteiam a assistência dos profissionais que atuam em um Centro de Atenção Psicossocial para atendimento de pacientes com transtornos, decorrentes do uso e dependência de substâncias psicoativas, na tentativa de subsidiar as ações dos demais serviços de saúde que atendem tais pacientes. Optou-se pela pesquisa qualitativa, seguindo a metodologia de estudo de caso; assim, nove profissionais foram entrevistados e o eixo norteador das questões foi referente à concepção de serviço e as ações assistenciais ali desenvolvidas. Os dados foram analisados à luz do conceito da representação social. Valendo-se dos temas emergentes foram elaboradas duas representações centrais da pesquisa: Concepção sobre o serviço de atenção psicossocial. Ações assistenciais aos pacientes com transtornos decorrentes de uso prejudicial e ou dependência de álcool e outras drogas. A análise final orienta-se para a compreensão de que o uso prejudicial do álcool e outras drogas decorrem de fatores multifacetados, propondo nessa linha de raciocínio a assistência psicossocial / The purpose of this study is to understand concepts and strategies that orient the assistance to professionals working in a Center of Psychosocial Attention to assist patients with disorders due to use and addiction of psychoactive substances, in the attempt to support actions of other health services which assist these patients. A qualitative research was our choice following the methodology of a case study. Nine professionals were interviewed and the interviews were based on questions aiming service and the assistance actions developed in the same institutions. Data were analyzed under the point of view of the social representation. Considering the emergent issue, two central representations of the research were elaborated: conception on the service of psychosocial attention. Assistance actions to patients with disorders due to the harmful use and/or addiction to alcohol and other drugs. The final analysis is oriented towards the comprehension that the harmful use of alcohol and other drugs are caused by varied factors, proposing a psychosocial assistance