Dissertations / Theses on the topic 'Intranasal administration of dopamine'

Recent research has indicated a causal link between oxytocin and spirituality. The present experiment sought to examine the effects of intranasal oxytocin (IN-OT) and absorption on mysticism and spirituality in a sensory deprivation setting. The results failed to find any main effects of IN-OT on mysticism, or on spirituality. Interaction effects were discovered however, where IN-OT interacted with absorption both on mysticism and spirituality. More specifically IN-OT undermined the association between absorption and outcomes that were observed in the placebo control condition. The results contradict the findings from the only previous experiment conducted on IN-OT and spirituality. The interaction effects align with previous research on IN-OT, suggesting an increase in suggestibility among low absorption scorers. These results motivate further research on the relation between oxytocin, absorption and spirituality, keeping the idea of suggestibility in mind.

Objetivos: Avaliar a segurança e a eficácia do midazolam intranasal (MIN) para sedação para tomografia em crianças, bem como a qualidade dos estudos radiológicos obtidos com esta técnica. Material e métodos: Entre dezembro de 2011 e julho de 2012, este estudo prospectivo avaliou o MIN como sedativo para crianças submetidas à tomografia sem acesso venoso. Após aprovação do Comitê de Ética em Pesquisa e consentimento dos responsáveis, 0,4 mg/kg de MIN foi administrado, sendo feita dose adicional de 0,1 mg/kg se o nível de sedação avaliado pela Escala de Sedação de Ramsay não fosse atingida após 15 minutos da primeira dose. Os desfechos relacionados à sedação incluíram tempo para sedação e para atingir os critérios de alta; parâmetros fisiológicos como oximetria de pulso e frequência cardíaca foram registrados a cada cinco minutos até a alta. A qualidade dos exames tomográficos foi avaliada quanto à presença de artefatos de imagem e movimento. Resultados: 60 eventos de sedação foram realizados em 58 pacientes. A idade média foi de 15,5 meses, sendo 90,9% dos exames tomográficos de crânio. O tempo médio para sedação foi de 15,2 minutos (5-40) e o tempo médio para atingir os critérios de alta foi de 74,7 minutos. Eventos adversos foram observados em 5 crianças (8,4%), incluindo reação paradoxal (3), tempo de recuperação prolongado (1) e vômitos (1). Apenas 4 pacientes (6,7%) não foram adequadamente sedados com MIN. Imagens consideradas excelentes, sem artefatos, foram obtidas em 56 (93,3%) sedações. Não houve eventos como bradicardia, hipoxemia ou hipotensão. Conclusões: O midazolam intranasal, administrado via atomizador nasal, é um método simples e não-invasivo para sedação segura, eficaz e previsível para crianças na obtenção de estudos tomográficos de qualidade<br>Objective: To evaluate the safety, efficacy and image quality of sedation with aerosolized intranasal midazolam for pediatric CT studies. Materials and Methods: Between December 2011 to May 2012, this prospective study evaluated aerosolized intranasal (AIN) midazolam as a sedative for CT of children without intravenous access. After IRB approval and parental consent, 0,4 mg/kg of AIN midazolam was administered, and repeated with 0.1 mg/kg if adequate sedation evaluated by Ramsay Sedation Scale not achieved in 15 minutes after the first dose. Sedation outcome variables which included time to achieve sedation, to meet discharge criteria and physiological vital signs of pulse oximetry and heart rate, were recorded every five minutes until discharge. The quality of CT images was reviewed and graded for presence of motion and imaging artifacts, Results: 60 sedation encounters were performed in 58 children. Mean age was 15.5 months, and 90.9% of CT scans were brain scans. Mean time to sedation was 15.2 minutes (range 5-40) and mean time to achieve discharge criteria was 74.7 minutes. Adverse events were recorded in 5 children (8.4%) that underwent sedation - paradoxical reaction (3), prolonged recovery time (1) and vomiting (1). Only 4 patients (6.7%) failed to sedate. Excellent CT imaging, with no artifacts, were obtained in 56 (93.3%) of sedation encounters. No adverse events like bradycardia, hypoxia or hypotension were documented. Conclusions: The aerosolized route of administration of midazolam is a simple and noninvasive approach for predictable, effective and safe sedation of children for quality CT imaging studies

Orientador: Ricardo de Lima Zollner<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas<br>Made available in DSpace on 2018-08-14T06:15:03Z (GMT). No. of bitstreams: 1 ZanellatoFabbri_Natalia_M.pdf: 1108663 bytes, checksum: 3362aa87243ab7742f6bb55df1f964e6 (MD5) Previous issue date: 2009<br>Resumo: Rinite alérgica é uma doença comum com ampla morbidade, que gera aumento considerável nos custos de tratamento médico, redução da produtividade no trabalho e absenteísmo escolar. A aplicação tópica de corticosteróides intranasal é amplamente reconhecida como primeira linha de tratamento antiinflamatório. Espera-se que a maioria dos sprays nasais prescritos como drogas de ação local, ainda não possuam patente permitindo o aumento de cópias genéricas desses medicamentos, levando a concorrência e redução de preço. Estudos de bioequivalência para sprays nasais estão ainda em discussão. Geralmente, os estudos para essa finalidade usa modelos de intervenção terapêutica a longo prazo, com altos custos para o paciente e longo tempo de duração. O objetivo deste trabalho foi mostrar a aplicabilidade da provocação nasal com histamina e rinomanometria em estudos de bioequivalência para sprays nasais. Trata-se de um estudo aberto, cruzado aleatorizado, utilizando dois períodos e duas seqüências para avaliar a equivalência farmacodinâmica entre duas formulações de sprays de dipropionato de beclometasona de manufaturamento distinto. Após estímulo nasal com histamina (0,5 mg/ml em ambas narinas), 25 voluntários saudáveis foram submetidos a rinomanometria anterior nos tempos 0; 15; 30 e 60 minutos para estabelecimento do fluxo, pressão e resistência basal de cada câmara nasal. Os voluntários foram submetidos à utilização do spray nasal com a droga teste (T) ou referência (R) de acordo com o esquema de randomização, e a área sobre a curva foi analisada. De acordo com os parâmetros estudados os resultados obtidos mostraram diferença significativa entre as duas formulações (p= 0,31) indicando a equivalência terapêutica entre as drogas T e R. Assim, estes resultados sugerem que provocação nasal com histamina em indivíduos saudáveis e o emprego da rinomanometria como método quantitativo das alterações provocadas nas câmaras nasais possam ser aplicáveis em estudos farmacodinâmicos de bioequivalência para sprays nasais<br>Abstract: Allergic rhinitis is a common condition with widespread morbidity, increased medical treatment costs, reduced work productivity and lost school days. Topically delivered intranasal corticosteroids are widely recognized to be the first-line anti-inflammatory treatment. It is expected that many of the most-prescribed nasal sprays for local action drugs will go off patent, allowing the increase of the generic copies of these medications, with more products competition and price reduction. The bioequivalence studies for nasal sprays are still in discussion. Usually the studies designs for this purpose use a long-term therapeutic intervention models using patients with high costs and time duration. This study was designed to demonstrate the feasibility of rhinomanometry in bioequivalence studies for nasal sprays. Study design, an open, randomized, crossover study, using two periods and two sequences to evaluate pharmacodynamic equivalence between two formulations of beclometasone dipropionate spray. After nasal challenge with histamine (0.5mg/ml, in both nostrils), 25 healthy volunteers were submited to an anterior rhinomanometry at the time 0; 15; 30 and 60 minutes building a baseline of flow, pression and resistance of nasal chamber. Then, the vontuteers were submited to nasal drug spray (Test (T) or Reference(R)), according to randomized schedule and the Are Under Curve (AUC0-t) analyzed. The results suggest the potential use of nasal histamine challenge and rhinomanometry evaluation in healthy subjects in bioequivalence studies for nasal sprays<br>Mestrado<br>Ciencias Basicas<br>Mestre em Clinica Medica

The nasal administration of drugs offers advantages over administration by intravenous injection. Drugs can be rapidly absorbed through the nasal mucosa, resulting in a rapid onset of action, and also avoiding degradation in the gastro-intestinal tract and first-pass metabolism in the liver. Targeting the brain via nasal administration offers potential for the development of new drug products. The olfactory cells are in direct contact with both the environment and the central nervous system (CNS). The olfactory pathway thus circumvents the blood-brain barrier (BBB), which prevents many systemically administered drugs from entering the brain. A literature study concerning the anatomy a nd physiology oft he nose, factors affecting the absorption of nasally administered drugs; different mechanisms to enhance nasal drug absorption as well as the general characteristics of prochlorperazine edisylate were performed. As a result of the literature study, it was concluded that the nasal route is suited for administration and absorption of prochlorperazine edisylate. The aim of this study was to compare the concentrations of prochlorperazine found in plasma and brain tissue of rats, after intravenous, oral and intranasal administration. Methods: In order to investigate the objective, a dose of 0.167 mg/kg prochlorperazine edisylate was administered intravenously (100 µI), nasally (50 µl) and 1.333 mg/kg was administered orally (100 µl). The concentrations were corrected for the different dosages in order to compare the respective bioavailabilities. The nasal bioavailability of prochlorperazine was investigated in a rat model: uptake in the brain tissue and plasma levels were compared after intravenous, oral and intranasal administration. A liquid-liquid extraction method for the quantitative determination of prochlorperazine in brain tissue and a solid-phase extraction method for the quantitative determination of prochlorperazine in plasma were used. The concentrations of prochlorperazine in plasma and brain tissue were measured with high performance liquid chromatography. Results: The results indicate that the nasal administration of prochlorperazine is an easy and workable alternative to intravenous injections, which may enhance patient compliance. Nasal absorption: The concentration-time profile achieved after nasal administration of prochlorperazine is similar to that achieved after intravenous administration. The absorption of prochlorperazine edisylate from the nasal cavity into the systemic circulation was rapid and almost complete. The AUC-values, used as an indication of the extent of absorption, for the intravenous (3371.47 ± 173.79 ng/ml/h), intranasal (2936.71 ± 189.65 ng/ml/h) and oral routes of administration (718.07 ± 42.74 ng/ml/h) were compared. Compared to the intravenous route of administration (100%), the nasal route showed an absolute bioavailability of 87.10% and the oral route 21.30%. A low oral bioavailability was achieved, as expected, due to degradation in the gastro-intestinal tract and firstpass metabolism in the liver. Uptake into the brain tissue: The concentration-time profiles of prochlorperazine in brain tissue showed no increased maximum concentrations of drug after nasal administration compared to intravenous administration. However, this concentration was retained longer after nasal administration compared to intravenous administration. No direct evidence for transfer along the olfactory pathway was shown with prochlorperazine. The intranasal/intravenous brain tissue concentration ratio exceeded one after 30 and 45 minutes after nasal administration at a pH of 6.6 and 4.65 respectively, indicating that after these time intervals the concentrations of nasally administered prochlorperazine in the brain tissue were higher than those after intravenous administration. Prochlorperazine concentrations in the brain tissue were significantly higher after nasal administration than after oral administration. Significant concentrations of prochlorperazine were found in the brain tissue as early as 5 minutes after nasal administration. The AUC-values after nasal (96745.32 ± 3649.65 ng/g/h), intravenous (90051.71 ± 6189.75 ng/g/h) and oral administration (12507.20 ± 1248.01 ng/g/h) indicated that the nasal/intravenous AUC ratio in brain tissue was found to be greater than one. Conclusion: Nasal administration of CNS-active anti-emetic drugs with low oral bioavailability could be used as an alternative for the intravenous route of administration. The lipophilic drug, prochlorperazine was rapidly and almost completely absorbed after nasal administration. These molecules appeared rapidly in the brain tissue. Although hard evidence of direct transfer from the nose remains elusive, the fact that a higher AUC-value was obtained after nasal than after intravenous administration was evidence enough that the olfactory route does contribute to the delivery of drugs to the brain after nasal administration.<br>Thesis (M.Sc. (Pharm.))--North-West University, Potchefstroom Campus, 2004.

Master of Science<br>Department of Clinical Science<br>Elizabeth Davis<br>Background: While there is a commercially-available vaccine for Streptococcus equi subsp. equi licensed for the intranasal route of administration, some equine practitioners are administering this vaccine orally despite a lack of evidence for its efficacy by this route of administration. Objectives: To compare systemic and local immune responses following intranasal or oral administration of the USDA-approved, live-attenuated Streptococcus equi subspecies equi vaccine (Pinnacle IN®, Zoetis, Florham Park, New Jersey). Study Design: Experimental, randomized clinical trial Methods: Eight healthy horses with low Streptococcus equi M protein (SeM) titers (<1:1600) were randomly assigned to an intranasal or oral two-vaccine series. SeM-specific serum immunoglobulins G (IgG) and A (IgA) and nasal secretion IgA were assessed using a commercially-available ELISA (Equine Diagnostic Solutions, LLC, Lexington, Kentucky) and a novel magnetic microsphere assay utilizing fluorescence. A general linear mixed models approach was used for statistical data analysis. Results: As expected, intranasal vaccinates showed substantial increases in both serum SeM-specific IgG and IgA levels post-vaccination (P=0.0006 and P=0.007, respectively). Oral vaccinates showed an increase in serum SeM-specific IgG post-vaccination (P=0.0150), though only one-third the magnitude of intranasal vaccinates. Oral vaccinates showed no evidence of change in SeM-specific IgA post-vaccination (P=0.15). Main Limitations: Changes in mucosal antibody responses were not identified in this study which may be related to small change in antibody response, timing of sample collection, or method of nasal secretion collection. Conclusions: Results indicate that intranasal or oral vaccine administration resulted in increased serum SeM-specific IgG, though the magnitude of response differed between routes.

Schizophrenia is a chronic enduring disorder ranked among the most debilitating mental illnesses (Mueser & McGurk, 2004; Tandon, Keshavan, & Nasrallah, 2008). Although it has been vigorously studied over the past century, the etiology and pathophysiology of schizophrenia remains largely unknown and currently available treatments, in the form of antipsychotics, are mainly unsatisfactory (Insel, 2010; Tandon et al., 2008). Schizophrenia is characterized by three broad types of symptoms: positive symptoms, negative symptoms, and cognitive deficits. While drugs currently available for the treatment of this disorder are effective for positive symptoms, negative symptoms (including social impairments) and cognitive deficits still remain mainly untreatable.(Keefe et al., 2007; Neill et al., 2010). Negative and cognitive symptoms are more pervasive, fluctuate less over time than psychotic symptoms and are strongly associated with poor psychosocial functioning in community living and work (Kasper & Resinger, 2003; Mueser & McGurk, 2004; Tandon, Nasrallah, & Keshavan, 2009). Although there have been rapid progress in the development of non-invasive technologies to study human brain structure and function in the last two decades, there are still substantial limitations in our ability to investigate details of the physiology and molecular biology of the human brain (Nestler & Hyman, 2010a). To that end, it is imperative to have carefully validated animal models for continued progress in our understanding of pathophysiology and in the development and screening of novel therapeutic agents in order to enhance functional recovery of patients (Davis et al., 2013; Neill et al., 2010). Neuropeptides have an important role in intracerebral signaling and might have the potential to be used as therapeutic agents in many psychiatric and neurological diseases(Bedse, Di Domenico, Serviddio, & Cassano, 2015; Erbaş, Çınar, Solmaz, Çavuşoğlu, & Ateş, 2015; Nishimura, Murayama, & Takahashi, 2015; Reglodi et al., 2015, 2015). Unfortunately, they usually can’t be administered systemically due to the elevated hydrophilicity and molecular weight that prevent them to overcame the blood brain barrier. Intranasal administration might be a promising and non-invasive way of administration of neuropeptides: this administration route enables highly hydrophilic and high molecular weight molecules to bypass the blood-brain barrier permitting them to reach the brain in a non-invasive way. This way has been demonstrated in in humans that permits the delivery of biologically effective concentrations of many peptides to the brain without eliciting significantly eventual systemic hormone-like side effects (Born et al., 2002); This route of administration was tested also in mice and rats (Born et al., 2002; Neumann, Maloumby, Beiderbeck, Lukas, & Landgraf, 2013)and, recently, was successfully used specifically in the context of behavioural studies in in mice (Huang et al., 2014). The aim of the studies described in this thesis is to investigate, through the use of clinically-relevant animal models of schizophrenia, the pharmacogenetics behavioral effects of intranasal administration of two different neuropeptides: - oxytocin (OXT), a neurohypophyseal peptide suggested to have beneficial effects in social behaviors (Meyer-Lindenberg, Domes, Kirsch, & Heinrichs, 2011; Striepens, Kendrick, Maier, & Hurlemann, 2011) and currently in clinical studies for mental disorders characterized by social behavioral alterations such as autism (Anagnostou et al., 2012; Guastella et al., 2010) and schizophrenia (Feifel et al., 2010). - CRF(6-33), a syntetic peptide designed to be an effective competitive antagonist for the binding between the endogenous CRF neuropeptide and its binding protein (CRFbp)(Sutton et al., 1995) suggested in preclinical ICV studies to be an effective pro cognitive agent with potential therapeutic applications (Stephen C. Heinrichs & Joppa, 2001; Stephen C. Heinrichs, 2003; Koob & Bloom, 1985) In previous work from our laboratory (Huang et al., 2014), we already implemented, for the first time, the use of intranasal oxytocin in C57BL/6J mice, checking different behavioral effects. Thus, we now tested the effects of intransal OXT in the schizophrenia-relevant dysbindin-1 knockout mouse model. Genetic variations of the dysbindin-1 gene (DTNBP 1) has been associated with susceptibility to schizophrenia (O’Tuathaigh et al., 2007; Ross, Margolis, Reading, Pletnikov, & Coyle, 2006; Straub et al., 2002) and severity of negative symptoms and cognitive dysfunction in schizophrenic patients (Burdick et al., 2007; DeRosse et al., 2006; Fanous et al., 2005; Straub et al., 2002). Thereafter, using a Dys knockout mutant mouse model, we demonstrated that both the heterozygous and homozygous knockout mice manifested a reduction in social interaction compared to wildtype mice. Similar social deficits in Dys mutant mice have been reported by other groups (Feng et al., 2008; Hattori et al., 2008). Interestingly, both chronic and acute intranasal OXT treatments were able to ameliorate the social deficits observed in the Dys knockout mice. These data suggest that intranasal oxytocin might be beneficial to subjects with genetic modifications relevant to schizophrenia, while administration to healthy subjects has not significant behavioral effect or may eventually be detrimental (Huang et al., 2014). This opens new ways of exploration in relationship to the beneficial effects of OXT treatment and its utility in the clinical setting. The data (aforementioned) correlate well with the molecular data available. As a matter of fact, OXT receptors are downregulated in WT subjects that are chronically IN –OXT treated, and even mutant mice, though treated with the same therapeutic protocol, show that the receptors are not subject to variation , compared to the control subjects treated with the vehicle substance only. Indeed, after being chronically treated, WT mice showed a decrease in social behaviour, while mutant mice (under an equivalent OXT treatment) showed an increase in social behaviour. For the CRF(6-33) part, in order to set the ground for future studies with genetically modified mice, disease-related mouse models and facilitate inter-laboratory comparisons, we first tested the effects of both chronic and acute intranasal CRF treatments in C57BL/6J mice. Considered that OXT did not show significant improvement in cognitive performance, we chose to target these functions with a different peptide that in previous intracerebroventricular studies had shown to improve cognitive functions (Behan et al., 1995; Eckart et al., 1999; S C Heinrichs et al., 1997; Stephen C Heinrichs & Koob, 2004; Stephen C. Heinrichs & Joppa, 2001; Stephen C. Heinrichs, 2003; Koob & Bloom, 1985; Lee, Lee, Wang, & Lin, 1993; Lee & Sung, 1989; Thompson, Erickson, Schulkin, & Rosen, 2004) had shown to improve cognitive functions . To asses cognitive functions we used a modified version of the 5-Choice Serial Reaction Time Task (5-csrtt), a rodent test designed as analog of the Continuous Performance Test (cpt) used to asses quantitatively attentional control in humans (Amitai & Markou, 2011). The modifications are intended to reduce the time needed to train the animals , reduce stressful manipulations as food restriction or single housing. Additionally, several new manipulations have been implemented to investigate various specific cognitive functions such as attention, broad monitoring / compulsivity, response disinhibition/ impulsivity, distractibility and processing speed. We were able to demonstrate that the intranasal administration of CRF(6-33) can produce selective behavioral effects in mice. In particular, acute administration was able to improve accuracy of responses and reduce impulsivity, while chronic administrations produced a delay in correct responses. Gene expression studies with real time PCR are starting to suggest that the CRF(6-33) is able to reach the brain, as both CRFr1 and CRFbp were altered following intranasal CRF(6-33) in different and specific brain areas (i.e. Hippocampus and Prefrontal Cortex ). Subsequent dose-response tests confirmed the ability of evocate behavioral effects with a much lower dose of CRF(6-33). Moreover, we discovered a rebound effect in impulsive behavior the day after administration of higher doses.. This detrimental effect was absent with the lowest doses that was still able to significantly reduce impulsive behavior. Lastly we tested ability of CRF(6-33) to ameliorate an impulsive phenotype in a genetic modified mouse model of schizophrenia. The chosen model was a double mutant for Dysbindin and for the receptor D2. The D2 receptor that had proven in previous test to have an increased impulsive behaviour. The dopamine D2 receptor (D2) gene is another important risk gene identified for schizophrenia. Functional genetic variants in the D2 gene have been found to be differently expressed in patients with schizophrenia (Kaalund et al., 2013) and might modulate schizophrenia-related phenotypes by modifying the ratio of the short isoform (D2S) to the long isoform (D2L) (Bertolino et al., 2009). Heterozigote mutant for D2L has an increased D2S (receptor D2 short form)/ D2L (receptor D2 long form) ratio. From preliminary data CRF(6-33) was not able to significantly affect impulsive behaviour in double heterozygote Dys +/- D2L +/- . Interestigly, in single heterozygote Dys +/- was observed a trend of increased impulsivity il CRF(6-33) group suggesting a detrimental interaction with Dys deficient genotype. From this studies we can conclude that both OXT than CRF(6-33) have the potential to be used for treatment, respectively of social and impulsivity deficits. For OXT so far we observed a positive interaction with Dys deficient genotype , that needs to be confirmed in other schizophrenia relevant mouse models of social deficits. CRF(6-33) so far has demonstrated only to improve social performance in WT mice. The preliminary study on mutated mouse,if confirmed, seems to suggest that it might worsen the phenotype in presence of certain genetic mutations. It should be important to define mechanisms of interaction of CRF(6-33) with genetics as to define when it could be positively used for therapy and when it shouldn’t in a view of a genetic driven personalized schizophrenia therapy.<br>La schizofrenia è un disturbo cronico duraturo classificato tra le malattie mentali più debilitanti (Mueser e McGurk, 2004; Tandon, Keshavan, e Nasrallah, 2008). Anche se è stato vigorosamente studiata nel corso dell'ultimo secolo, l'eziologia e fisiopatologia della schizofrenia rimane in gran parte sconosciute e attualmente i trattamenti disponibili, in forma di antipsicotici, sono fondamentalmente insoddisfacente (Insel, 2010; Tandon et al., 2008). La schizofrenia è caratterizzata da tre grandi tipi di sintomi: sintomi positivi, i sintomi negativi e deficit cognitivi. Mentre i farmaci attualmente disponibili per il trattamento di questo disturbo sono efficaci per i sintomi positivi, sintomi negativi (compresi i deficit sociali) e deficit cognitivi rimangono principalmente incurabile. (Keefe et al, 2007;.. Neill et al, 2010). Sintomi negativi e cognitivi sono più pervasivi, fluttuano meno nel tempo dei sintomi psicotici e sono fortemente associati con scarso funzionamento psicosociale nella vita comunitaria e di lavoro (Kasper & Resinger, 2003; Mueser & McGurk, 2004; Tandon, Nasrallah, e Keshavan, 2009) . Anche se ci sono stati rapidi progressi nello sviluppo di tecnologie non invasive per studiare la struttura del cervello umano e il suo funzionamento negli ultimi due decenni, ci sono ancora limitazioni sostanziali nella nostra capacità di indagare i dettagli della fisiologia e della biologia molecolare del cervello umano (Nestler & Hyman, 2010a). A tal fine, è indispensabile avere modelli animali accuratamente convalidati per proseguire i progressi nella nostra comprensione della fisiopatologia e nello sviluppo e lo screening di nuovi agenti terapeutici al fine di migliorare il recupero funzionale dei pazienti (Davis et al, 2013;. Neill et al ., 2010). Neuropeptidi hanno un ruolo importante nella segnalazione intracerebrale e potrebbe avere il potenziale per essere utilizzati come agenti terapeutici in molte malattie psichiatriche e neurologiche (Bedse, Di Domenico, Serviddio, e Cassano, 2015; Erbas, Cinar, Solmaz, Cavusoglu, e Ates, 2015 , Nishimura, Murayama, e Takahashi, 2015;. Reglodi et al, 2015, 2015). Purtroppo, di solito non possono essere somministrati per via sistemica a causa della idrofilia elevata e peso molecolare che impediscono loro di superamento del la barriera ematoencefalica. La via di somministrazione intranasale potrebbe essere un modo promettente e non invasivo di somministrazione di neuropeptidi: questa via di somministrazione permette molecole altamente idrofile e ad alto peso molecolare per bypassare la barriera emato-encefalica, consentendo loro di raggiungere il cervello in modo non invasivo. In questo modo è stato dimostrato in negli esseri umani che permette la consegna delle concentrazioni biologicamente efficaci di molti peptidi al cervello, senza suscitare in modo significativo eventuali effetti collaterali ormonali sistemici (Born et al., 2002); Questa via di somministrazione è stata testata anche in topi e ratti (Nato et al., 2002; Neumann, Maloumby, Beiderbeck, Lukas, e Landgraf, 2013) e, recentemente, è stato utilizzato con successo in particolare nel contesto degli studi comportamentali nei topi ( Huang et al., 2014). Lo scopo dello studio descritto in questa tesi è di indagare, attraverso l'uso di modelli animali clinicamente rilevanti per la schizofrenia, la farmacogenetica degli effetti comportamentali della somministrazione intranasale di due neuropeptidi differenti: - Ossitocina (OXT), un peptide neuroipofisario suggerito avere effetti benefici in comportamenti sociali (Meyer-Lindenberg, Domes, Kirsch, e Heinrichs, 2011; Striepens, Kendrick, Maier, e Hürlemann, 2011) e attualmente in studi clinici per i disturbi mentali caratterizzata da alterazioni comportamentali sociali come l'autismo (Anagnostou et al, 2012;.. Guastella et al, 2010) e la schizofrenia (Feifel et al., 2010). - (. Sutton et al, 1995) CRF (6-33), un peptide sintetico progettato per essere un antagonista competitivo efficace per il legame tra il neuropeptide CRF endogena e la sua proteina legante (CRFbp) ha suggerito in studi preclinici ICV ad essere un efficace agente pro cognitivo con potenziali applicazioni terapeutiche (Stephen C. Heinrichs e Giaffa, 2001; Stephen C. Heinrichs, 2003; Koob & Bloom, 1985) In un precedente lavoro del nostro laboratorio (Huang et al., 2014), abbiamo già implementato, per la prima volta, l'uso di ossitocina intranasale in C57BL / 6J, controllando effetti comportamentali diversi. Così, ora abbiamo testato gli effetti di OXT intransale nel modello di topo dysbindin-1 knockout rilevante per schizofrenia. Variazioni genetiche del dysbindin-1 gene (DTNBP 1) è stato associato con la suscettibilità alla schizofrenia (O'Tuathaigh et al, 2007;. Ross, Margolis, lettura, Pletnikov, & Coyle, 2006;. Straub et al, 2002) e gravità dei sintomi negativi e disfunzioni cognitive nei pazienti schizofrenici (Burdick et al, 2007;. DeRosse et al, 2006;.. Fanous et al., 2005; Straub et al, 2002). Successivamente, utilizzando un modello di topo knockout mutante Dys, abbiamo dimostrato che sia i topi knockout eterozigoti e omozigoti manifestano una riduzione nell'interazione sociale rispetto ai topi di wild type. Deficit sociali simili a Dys topi mutanti sono stati segnalati da altri gruppi (Feng et al, 2008;. Hattori et al., 2008). È interessante notare che entrambi i trattamenti intranasale OXT, cronici e acuti, sono stati in grado di migliorare i deficit sociali osservati nei topi knockout Dys. Questi dati suggeriscono che intranasale di ossitocina potrebbe essere utile ai soggetti con modificazioni genetiche rilevanti per la schizofrenia, mentre la somministrazione a soggetti sani non ha significativo effetto comportamentale o alla fine può essere dannoso (Huang et al., 2014). Questo apre nuove vie di esplorazione in relazione agli effetti benefici del trattamento OXT e la sua utilità in ambito clinico. I dati (di cui sopra) correlano bene con i dati molecolari disponibili. È un dato di fatto, recettori OXT sono inibiti in soggetti WT che sono cronicamente IN -OXT trattati, e persino topi mutanti, anche se trattata con lo stesso protocollo terapeutico, mostrano che i recettori non sono soggetti a variazioni, rispetto ai soggetti di controllo trattati con solo la sostanza veicolo. Infatti, dopo essere stati trattati cronicamente, WT topi hanno mostrato una diminuzione del comportamento sociale, mentre i topi mutanti (nel quadro di un trattamento OXT equivalente) hanno mostrato un aumento dei comportamenti sociali. Per il CRF (6-33) parte, al fine di impostare le basi per futuri studi con i topi geneticamente modificati, modelli murini legati alla malattia e facilitare il confronto tra laboratori, in primo luogo abbiamo testato gli effetti di entrambi i trattamenti intranasale CRF cronici e acuti in C57BL / 6J. Considerato che OXT non ha mostrato un significativo miglioramento delle prestazioni cognitive, abbiamo scelto di indirizzare queste funzioni con un peptide diverso che in precedenti studi intracerebroventricolare aveva dimostrato di migliorare le funzioni cognitive (Behan et al, 1995;.. Eckart et al, 1999; SC Heinrichs et al, 1997;. Stephen C Heinrichs & Koob, 2004; Stephen C. Heinrichs e Giaffa, 2001; Stephen C. Heinrichs, 2003; Koob & Bloom, 1985; Lee, Lee, Wang, e Lin, 1993; Lee & Sung , 1989; Thompson, Erickson, Schulkin, e Rosen, 2004) aveva dimostrato di migliorare le funzioni cognitive. Per asini funzioni cognitive abbiamo utilizzato una versione modificata del 5 choice serial reaction time task (5-csrtt), un test per roditori progettato come analogo del Continuous Performance Test (cpt) utilizzato per testare quantitativamente il controllo dell'attenzione negli esseri umani (Amitai & Markou , 2011). Le modifiche sono destinate a ridurre il tempo necessario per addestrare gli animali, ridurre manipolazioni stressanti come restrizione alimentare o allevamento isolato. Inoltre, diverse nuove manipolazioni sono state implementate per studiare varie specifiche funzioni cognitive quali l'attenzione, broad monitoring / compulsività, risposta disinibizione / impulsività, distraibilità e velocità di elaborazione. Siamo stati in grado di dimostrare che la somministrazione intranasale di CRF (6-33) in grado di produrre effetti comportamentali selettivi nei topi. In particolare, la somministrazione acuta è stata in grado di migliorare la precisione delle risposte e ridurre l'impulsività, mentre le amministrazioni croniche hanno prodotto un ritardo nelle risposte corrette. Gli studi di espressione genica con real time PCR stanno iniziando a suggerire che il CRF (6-33) è in grado di raggiungere il cervello, poiché l’espressione di molecole come CRFR1 e CRFbp sono state modificate in seguito intranasale CRF (6-33) in diverse e specifiche aree cerebrali (cioè Hippocampus e corteccia prefrontale). Test dose-risposta successivi hanno confermato la possibilità di evocare effetti comportamentali con una dose molto più bassa di CRF (6-33). Inoltre, abbiamo scoperto un effetto di rimbalzo nel comportamento impulsivo il giorno dopo la somministrazione di dosi più elevate .. Questo effetto negativo è stato assente con le dosi più basse che ancora sono in grado di ridurre in modo significativo il comportamento impulsivo. Infine abbiamo testato la capacità di CRF (6-33) di migliorare un fenotipo impulsivo in un modello genetico del topo modificato della schizofrenia. Il modello scelto è stato un doppio mutante per Dysbindin e per la D2 recettore. Il recettore D2 che aveva dimostrato in prova precedente per avere una maggiore comportamento impulsivo. Il gene recettore D2 della dopamina (D2) è un altro importante gene rischi identificati per la schizofrenia. Varianti genetiche funzionali nel gene D2 sono stati trovati per essere espressi in modo diverso nei pazienti con schizofrenia (Kaalund et al., 2013) e possono modulare fenotipi correlato alla schizofrenia modificando il rapporto tra la breve isoforma (D2S) alla lunga isoforma (D2L ) (Bertolino et al., 2009). Mutante Heterozigote per D2L ha una maggiore rapporto D2S (recettore D2 forma breve) / D2L (recettore forma estesa D2). Dai dati preliminari CRF (6-33) non è stato in grado di incidere in modo significativo il comportamento impulsivo in Dys doppi eterozigoti +/- D2L +/-. È interessante notare che, in Dys singoli eterozigote +/- è stata osservata una tendenza di aumento impulsività il CRF (6-33) del gruppo suggerisce una interazione dannoso con Dys genotipo carente. Da questi studi si può concludere che sia OXT di CRF (6-33) hanno il potenziale per essere utilizzato per il trattamento, rispettivamente, di deficit sociali e impulsività. Per OXT finora abbiamo osservato una interazione positiva con il genotipo deficitario per Dys, che ha bisogno di essere confermata in altri modelli murini di schizofrenia rilevanti di deficit sociali. CRF (6-33) finora ha dimostrato solo per migliorare le prestazioni sociali in topi WT. Lo studio preliminare sul mouse mutato, se confermato, sembra suggerire che potrebbe peggiorare il fenotipo in presenza di alcune mutazioni genetiche. E’ importante quindi definire meccanismi di interazione di CRF (6-33) con la genetica per definire quando potrebbe essere utilizzato positivamente per la terapia e quando non dovrebbe in una visione di un regime terapeutico per la schizofrenia personalizzato e direzionato da test genetici.

Bakgrund: Ett vanligt symtom inom ambulanssjukvården är smärta, vilken ofta kan förvärra den smärta som redan upplevs av patienten, då de nödvändiga förflyttningarna genomförs av besättningen till bår och ambulans. Det är sjuksköterskorna i ambulansbesättningen som gör bedömningen huruvida smärtlindring krävs akut eller inte under transporten till sjukhus. Syfte: Syftet med studien var att studera patienters upplevelser av intranasal (i.n) smärtlindring jämfört med intravenös (i.v) smärtlindring inom ambulanssjukvården i Stockholm. Metod: Studien är en empirisk studie med deskriptiv design. Huvudresultat: Studien visade endast signifikant skillnad i området där upplevelsen av oro och/eller rädsla mättes i samband med i.n kontra i.v smärtlindringsmetod. Det var fler som upplevde en negativ aspekt i form av oro och eller rädsla i den grupp som fått i.n smärtlindring. Inga andra signifikanta skillnader i upplevelsen av varken, nöjdhet av smärtlindringens effekt, själva sättet att erhålla läkemedlet eller upplevelse av biverkningar påvisades. En jämförelse mellan de rapporterade biverkningarna visade att illamående var den vanligaste biverkningen och den var vanligast i den grupp som erhållit i.n smärtlindring. Slutsats: Med bakgrund av detta resultat kan man ifrågasätta om administreringssättet är avgörande kring effekten och upplevelsen av positiva eller negativa aspekter av smärtbehandlingen.<br>Background: A common symptom of ambulance care is pain, which can often exacerbate the pain already experienced by the patient, as the necessary movements are carried out by the crew to the stern and ambulance. It is the nurses in the ambulance crew who assess whether pain relief is required urgently or not during transport to hospitals.   Objective: The purpose of this study was to study patient´s experiences of intranasal (i.n) pain relief compared to intravenous (i.v) pain relief within the ambulance service in Stockholm area.    Method: The study is an empirical study with a descriptive design.   Main results: The study showed that the only significant difference in the area where the experience of anxiety and/or fear was measured in connection with i. n versus i. v. pain relief method. It was more that experienced a negative aspect in the form of anxiety and or fear in the group that received in n pain relief. No other significant differences in the experience of pain relief, satisfaction of neither the ring's power, the way to obtain the medicine or experience side effects was demonstrated. A comparison of the reported showed that the most common side effect was nausea, and it was most common in the group that received in n pain relief.   Conclusion: Based on this result, one can question whether the mode of administration is crucial for the effect and experience of positive or negative aspects of pain management.

INTRODUÇÃO: O midazolam é uma imidazobenzodiazepina usada para induzir o sono, produzir sedação antes de procedimentos dolorosos e no tratamento do estado de mal epiléptico. Seu uso pela via intranasal proporciona um rápido início de ação e esta via, em muitos casos, pode substituir as vias endovenosa e intramuscular, mais invasivas. Assim, o midazolam intranasal tem sido sugerido no tratamento extra-hospitalar de crises epilépticas e ataques de pânico. Por outro lado, os benzodiazepínicos possuem um potencial para serem abusados, principalmente em usuários de outras drogas. OBJETIVO: o presente estudo objetivou verificar o potencial de abuso do midazolam intranasal numa população experiente no uso intranasal de substâncias - abusadores de cocaína aspirada. MÉTODOS: Foram estudados 31 voluntários abusadores ou dependentes de cocaína e 34 controles saudáveis, subdivididos em quatro grupos: Abusadores de Cocaína (N = 16) e Voluntários Saudáveis (N= 17) que receberam midazolam (0,5 mg de hidrocloreto de midazolam em cada narina), e Abusadores de Cocaína (N = 15) e Voluntários Saudáveis (N = 17) que receberam o mesmo volume de um placebo ativo. As variáveis de resposta foram a Apreciação da Substância (AS) e a Vontade de Repetir o Uso da Substância (VR), avaliadas através de escalas analógicas visuais. RESULTADOS: A análise de perfis para medidas repetidas das variáveis de resposta mostrou um efeito significante da variável Tempo sobre AS (F[5;57] =3,947, p=0,004) e VR (F[5;57] =3,311, p=0,011). A variável Grupo (Abusadores de Cocaína x Voluntários Saudáveis) também teve um impacto sobre as variáveis de resposta AS e VR, sendo que os Abusadores de Cocaína tiveram pontuações mais altas tanto em AS (F[5;57] = 4,946, p = 0,030) quanto em VR (F[5;57] =5,229, p=0,026). Numa análise de regressão linear para investigar os efeitos do humor - medidos através de uma Escala Visual Analógica do Humor (VAMS) - sobre as variáveis de resposta AS e VR, os Abusadores de Cocaína apresentaram escores maiores que os Voluntários saudáveis tanto para AS (t = 3,37; p = 0,001) quanto para VR (t = 5,607; p = 0,011). Observou-se, também, um efeito dos fatores VAMS 16 (MAIS DEPRIMIDO MAIS EUFÓRICO; t = 4,28; p < 0,001) e VAMS 12 (MAIS EXCITADO MAIS RELAXADO; t = 2,66; p = 0,010), sobre a variável de resposta AS (R2 = 0,32): níveis maiores de euforia e relaxamento predisseram uma maior Apreciação da Substância instilada. O fator VAMS 16 (MAIS DEPRIMIDO MAIS EUFÓRICO) teve um efeito também sobre a variável de resposta VR (t = 3,65, p < 0,001; R2 =0,24): maior euforia predisse maior vontade de repetir o uso da substância. Finalmente, uma análise de regressão linear utilizando-se AS como variável explicativa e VR como variável de resposta resultou que quanto maior a apreciação positiva da substância, maior era a vontade de repetir seu uso (F = 108, 517; p < 0,001; R2 = 0,65). CONCLUSÕES: Corroborando estudos anteriores, observou-se que sensações como relaxamento e euforia correlacionam-se com o potencial de abuso de uma substância e uma maior apreciação positiva dos efeitos de uma substância correlaciona-se com uma maior vontade de usá-la novamente. Por outro lado, conclui-se que via intranasal em si aumentaria a probabilidade de abuso em usuários de substâncias intranasais. Consequentemente, uma eventual produção e comercialização para uso intranasal de uma substância com potencial de abuso deverá levar em conta este risco adicional para populações usuárias de drogas<br>INTRODUCTION: Midazolam is an imidazobenzodiazepine used for sleep induction, for sedation before painful procedures and in the treatment of status epilepticus. When it is administered through the intranasal route, it has a fast beginning action and this route can many times be substituted for the more invasive intravenous and intramuscular routes. Therefore, intranasal midazolam has been uggested for the community management of pileptic seizures and panic attacks. On the other side, benzodiazepines display abuse liability, particularly in substance abusers. OBJECTIVE: The present study aimed at examining the abuse liability of intranasal midazolam in a population experienced with the intranasal abuse of substances, namely snorted cocaine abusers. METHODS: Thirty-one subjects with diagnoses of snorted-cocaine abuse or dependence have been studied, divided in four groups: Cocaine-Abusers (n = 16) and Healthy Volunteers (n = 17) that received midazolam (0.5 mg midazolam hydrochloride in each nostril), and Cocaine-Abusers (n = 15) and Healthy Volunteers (n = 17) that received the same volume of an active placebo. Response variables were Substance Liking (SL) and the Desire to Take the Substance Again (SA), assessed through visual analogue scales. RESULTS: Profile analysis for repeated measures of the response variables showed a significant effect of Time over both SL (F[5,57] =3.947, p=0.004) and SA (F[5;57] =3.311, p=0.011). Group had also a significant effect, in that Cocaine Abusers scored higher in both SL (F[5,57] = 4.946, p = 0.030) and SA (F[5;57] =5.229, p=0.026). In a linear regression analysis examining the effects of mood (measured through Visual Analogic Mood Scales VAMS) over the response variables SL and SA, Cocaine Abusers displayed higher scores than Healthy Volunteers at both SL (t = 3.37; p = 0.01) and SA (t = 5.607; p = 0.011). It was also found that variables VAMS 16 (MORE DEPRESSED MORE EUPHORIC; t = 4.28; p < 0.001) and VAMS 12 (MORE EXCITED MORE RELAXED; t = 2.66; p = 0.010) had an effect over response variable SL (R2 = 0.32): higher euphoria and relaxation scores predicted more liking of the administered substance. Factor VAMS 16 (MORE DEPRESSED MORE EUPHORIC) had also an effect over response variable SA (t = 3.65, p < 0.001; R2 =0.24): more euphoria predicted more desire to take the drug again. Finally, in a linear regression with SL as explaining variable and SA as response variable, it was found that higher SL predicted a higher SA (F = 108.517; p < 0.001; R2 = 0.65). CONCLUSIONS: Corroborating previous findings in literature, it was observed that feelings of relaxation and euphoria after the administration of an intranasal substance are correlated with higher abuse liability and that subjects who report more liking of a substance do also report more desire to take it again. On the other hand, it is concluded that the intranasal route might per se increase the probability of abuse in intranasal-substance users. Therefore, the production and marketing for intranasal use of a substance with abuse liability should take into account this additional risk for intranasal drug abusing populations.

Cariprazine is a novel dopamine (DA) and serotonin (5-HT) partial agonist with an in vitro receptor affinity profile that endows it with the potential to be used successfully in the treatment of both unipolar and bipolar disorders. The objective of this study was to determine whether in vitro findings with cariprazine lead to functional alterations of monoamine systems in the intact rat brain. In vivo electrophysiological recordings were carried out in male Sprague-Dawley rats under chloral hydrate anesthesia. Dorsal raphé nucleus (DRN), locus coeruleus (LC), and hippocampus cornu ammonis region 3 (CA3) pyramidal neurons were recorded and cariprazine was administered systemically by intravenous injection or locally through iontophoresis. In the DRN, cariprazine induced a complete inhibition of the firing of 5-HT neurons, which was fully reversed by the selective 5-HT1A antagonist WAY100.635. In the LC, the inhibitory effect of the preferential 5-HT2A agonist 2,5-dimethoxy-4-iodoamphetamine (DOI) was reversed by cariprazine with an ED50 value of 67 µg/kg, i.v., and it did not block the inhibitory effect of the α2-adrenergic agonist clonidine. In the hippocampus, when cariprazine was administered by iontophoresis, it inhibited the firing of pyramidal neurons, but it did not dampen the suppressant effect of 5-HT. These results indicate that, in vivo, cariprazine acts as a 5-HT1A agonist in the DRN, as an antagonist on 5-HT2A receptors controlling the firing of NE neurons, and is a full agonist at 5-HT1A receptors located on pyramidal neurons of the hippocampus. The modulatory actions of cariprazine on the 5-HT and NE systems may contribute to its reported effectiveness in depressive episodes.

High grade gliomas remain a devastating disease, for which a curative therapy is virtually absent. The high medical need is unmet by novel treatment strategies and advances in chemo-and radiotherapy. Patients diagnosed for GBM face a median survival of 15 months after maximal standard-of-care therapy, and relapse is often observed due to micro-metastasis in the direct environment of resection. In part, current treatment modalities such as chemo-and immunotherapy are hampered in their efficacy due to the specialized TME. This area is adequately equipped to withstand the cytotoxic attack of chemo- and immunotherapy. Therefore, we hypothesized that modulation of the TME could decrease these defense mechanisms, and increase susceptibility to tumor lysis.In this respect, we focused on Gal-1 as an ideal target to modulate the TME in the context of GBM. Gal-1 exerts multiple tumor promoting functions. From pre-clinical research, we have learned that Gal-1 is an important mediator for the proliferation and migration of tumor cells, moreover Gal-1 could also promote angiogenesis in the TME, providing nutrients and oxygen for GBM to grow. Gal-1 also maintains the inherent defense mechanisms to chemo and immunotherapy. Gal-1 is crucial for the resistance mechanisms to TMZ by altering the EPR stress response. Moreover, and most important for our purposes, Gal-1 is also a crucial immune suppressor in the TME, which can induce apoptosis in activated T cells, and recruit Tregs. To target Gal-1 in the TME would be clinically most relevant if this could be performed via a non-invasive treatment modality. Therefore, we developed a nanoparticle complex that could deliver siGal-1 from the nasal cavity directly to the CNS, and even the TME. This nose-to-brain delivery bypasses systemic routes, with a higher (and more selective) local bioavailability in the CNS. The major pharmaceutical excipient in this nanoparticle complex consists of chitosan polymers. These polymers are highly interesting agents to promote nose-to-brain delivery due their muco-adhesive and epithelial barrier modulation properties. When applying these particles in vitro on GBM cells, a solid decrease of Gal-1 was noted, and the epithelial modulatory properties were confirmed. Furthermore, we observed a rapid transport from the nasal cavity to the brain upon intranasal administration of a highly-concentrated chitosan nanoparticle siGal-1 suspension and we could even observe the sequence-specific cleavage of Gal-1 mRNA, and a decrease of Gal-1 in the TME. This Gal-1 reduction could modulate the TME from immune suppression to immune activation, as demonstrated by decrease in suppressor cells, and increased stage of activation in rejective immune cells. Moreover, due to decreased Gal-1, also angiogenesis was alleviated, and a reduced size in vasculature was observed, mimicking a morphological vessel normalisation. Reversing the immune and vascular contexture of the TME by Gal-1 reduction seemed a prerequisite to increase the efficacy of TMZ, DC vaccination and PD-1 blocking. In combination experiments, we noticed that siGal-1 on top of these treatments, could further increase the efficiency of chemo and immunotherapy. The findings presented in this thesis can serve as a proof of concept for the feasibility to modulate and re-orchestrate the TME of GBM via intranasal administration. The intranasal administration of siGal-1 could represent a valuable clinically translational treatment to increase the efficiency of chemo- and immunotherapy for GBM patients. In our research facilities, a phase 0 as a first-in-human trial is actively pursued.<br>Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie)<br>info:eu-repo/semantics/nonPublished

Submitted by Erika Demachki (erikademachki@gmail.com) on 2017-04-05T21:16:41Z No. of bitstreams: 2 Dissertação - Joji Sado Filho - 2017.pdf: 5994690 bytes, checksum: 2a60faf14b700ecba0cd47970099e8f8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)<br>Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-04-06T15:04:55Z (GMT) No. of bitstreams: 2 Dissertação - Joji Sado Filho - 2017.pdf: 5994690 bytes, checksum: 2a60faf14b700ecba0cd47970099e8f8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)<br>Made available in DSpace on 2017-04-06T15:04:55Z (GMT). No. of bitstreams: 2 Dissertação - Joji Sado Filho - 2017.pdf: 5994690 bytes, checksum: 2a60faf14b700ecba0cd47970099e8f8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-13<br>Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq<br>During dental treatment of children, basic or advanced behavior guidance techniques can be used. Advanced techniques, non-pharmacological (protective stabilization) or pharmacological (sedation and general anesthesia), are indicated when there is no success with the basic techniques. Regarding advanced techniques, sedation has been increasingly widespread in pediatric dentistry, although the evidence is still weak about which sedative regime provides the greatest comfort and the lowest risk. In view of this need to identify the best sedative regimen, the objective of this randomized, masked, controlled, parallel-design trial was to evaluate the effectiveness of intranasal sedation with Midazolam and Ketamine in behavioral control of preschool children undergoing dental treatment. Eighty-four preschoolers, aged between 1.5 and 6 years old, with dental caries and non-cooperative behavior in previous dental treatment, were randomized into three groups: (1) Midazolam and Ketamine intranasally (MKI); (2) Midazolam and oral ketamine (MKO); (3) Oral midazolam. In all groups, the sedative was administered by a pediatrician or anesthesiologist, and the children received restorative treatment under local anesthesia and rubber dam isolation. Dental treatment was performed by pediatric dentists who, in the end, classified child behavior through the Frankl scale. The training sessions were videotaped, and the videos were then analyzed by trained and calibrated researchers to assess behavior using the Ohio State University Behavioral Rating Scale (OSUBRS) scale. The need to discontinue care and to use protective stabilization was recorded. The data obtained were organized and analyzed in the software Statistical Package for Social Sciences (SPSS). The statistical analysis involved the description of the data and bivariate tests, considering a level of significance of 5%. Considering as a parameter for sedation success the "positive" or "definitely positive" behavior verified with the Frankl scale, the following rates were observed: MKI - 50.0%; MKO - 64.3% and MO - 28.6%. Success rates differed significantly between MKO compared to MKI and MO (P = 0.03). When comparing the groups in relation to the behavior evaluated by the OSUBRS scale, no statistically significant difference was observed in the frequency of "quiet" (P = 0.22), "movement without crying" (P = 0.69) and "combative (P = 0.30). The groups did not differ in assessing the number of cases in which there was a "quiet" behavior in at least 51.7% (median) of the session duration (P = 0.27) and in the number of cases in which the child was "combative" "In at least 24.5% (median) of the session duration (P = 0.65). The need to suspend treatment (P = 0.69) and use protective stabilization (P = 0.14) did not differ significantly among the three groups. The results of behavior evaluation using the Frankl and OSUBRS scale were strongly correlated (rho = -0.84, P0.01). From these results, it was concluded that the sedative protocol MKO presented a higher success rate in behavioral control when compared to the other groups (MKI and MO). The combination of Midazolam and Ketamine, regardless of route of administration, was more effective in controlling behavior than Midazolam given alone. Therefore, the use of the combination of Midazolam and Ketamine, both orally and intranasally, is an effective alternative for controlling the behavior of non-collaborating preschoolers.<br>Durante o atendimento odontológico de crianças, podem ser utilizadas técnicas básicas ou avançadas de controle do comportamento. As técnicas avançadas, não farmacológicas (estabilização protetora) ou farmacológicas (sedação e anestesia geral), são indicadas quando não há sucesso com as técnicas básicas. Em relação às técnicas avançadas, a sedação tem sido cada vez mais difundida na odontopediatria, embora as evidências ainda sejam fracas sobre qual regime sedativo proporciona o maior conforto e menor risco. Diante dessa necessidade de se identificar o melhor regime sedativo, o objetivo deste ensaio clínico randomizado, mascarado, controlado e de delineamento paralelo foi avaliar a eficácia da sedação intranasal com Midazolam e Cetamina no controle comportamental de crianças pré-escolares submetidas a atendimento odontológico. Oitenta e quatro pré-escolares, com idade entre 1,5 e 6 anos, com cárie dentária e comportamento não colaborador em atendimentos odontológicos anteriores, foram randomizados em três grupos: (1) Midazolam e Cetamina por via intranasal (MKI); (2) Midazolam e Cetamina por via oral (MKO); (3) Midazolam por via oral. Em todos os grupos, o sedativo foi administrado por um médico pediatra ou anestesiologista, e as crianças receberam tratamento restaurador sob anestesia local e isolamento absoluto. O tratamento odontológico foi realizado por odontopediatras que, ao final, classificaram o comportamento infantil por meio da escala de Frankl. As sessões de atendimento foram filmadas e, posteriormente, os vídeos foram analisados por pesquisadores treinados e calibrados a fim de se avaliar o comportamento usando a escala Ohio State University Behavioral Rating Scale (OSUBRS). A necessidade de suspender o atendimento e de utilizar estabilização protetora foi registrada. Os dados obtidos foram organizados e analisados no software Statistical Package for Social Sciences (SPSS). A análise estatística envolveu a descrição dos dados e testes bivariados, considerando-se um nível de significância de 5%. Considerando-se como parâmetro para sucesso da sedação o comportamento “positivo” ou “definitivamente positivo” verificado com a escala de Frankl, foram observadas as seguintes taxas: MKI – 50,0%; MKO – 64,3% e MO – 28,6%. As taxas de sucesso diferiram significativamente entre MKO em comparação com MKI e MO (P = 0,03). Ao se comparar os grupos em relação ao comportamento avaliado pela escala OSUBRS, não foi observada diferença estatisticamente significativa na frequência de comportamentos “quieto” (P=0,22), “movimento sem choro” (P=0,69) e “combativo” (P=0,30). Os grupos não diferiram ao se avaliar o número de casos em que houve comportamento “quieto” em pelo menos 51,7% (mediana) da duração da sessão (P=0,27) e número de casos em que a criança ficou “combativa” em, pelo menos, 24,5% (mediana) da duração da sessão (P=0,65). A necessidade de suspender o atendimento (P=0,69) e de utilizar estabilização protetora (P=0,14) não diferiu significativamente entre os três grupos. Os resultados da avaliação do comportamento por meio da escala de Frankl e da OSUBRS foram fortemente correlacionados (rho = -0,84, P≤0,01). A partir destes resultados conclui-se que o protocolo sedativo MKO apresentou maior taxa de sucesso no controle comportamental, quando comparado aos outros grupos (MKI e MO). A combinação de Midazolam e Cetamina, independentemente da via de administração, foi mais eficaz para o controle do comportamento que o Midazolam administrado isoladamente. Sendo assim, o uso da associação Midazolam e Cetamina, tanto por via oral, quanto intranasal, é uma alternativa eficaz para o controle do comportamento de pré-escolares não colaboradores.

L’augmentation préoccupante de la résistance aux antibiotiques de Pseudomonas aeruginosa (PA) nécessite la recherche de thérapies alternatives telles que les Lactobacillus. Dans cette thèse, différents travaux ont été réalisés :1) Sur un modèle murin de pneumonie aigüe à PA, nous avons démontré l’effet bénéfique de l’administration intratrachéale d’un mélange de 3 souches de Lactobacillus provenant de lait ou de la cavité orale de patients sains.2) Nous avons ensuite prospectivement étudié la population en Lactobacillus des expectorations de patients atteints de mucoviscidose (CF). La prévalence moyenne de portage était de 61%.3) Parmi ces Lactobacillus issus de l’écosystème respiratoire des patients CF, nous avons constitué deux mélanges de 3 souches de Lactobacillus, sélectionnées pour leur activité anti-élastolytique et antipyocyanine in vitro. L’administration intranasale de ces mélanges à des souris C57Bl/6, 18h avant leur infection par PAO1, améliore significativement la survie à 7 jours des souris, ainsi que la clairance pulmonaire en PAO1 à 24h post-infection. Une diminution significative du recrutement pulmonaire des neutrophiles et des cytokines proinflammatoires était observée, associée à une augmentation de celle en IL-10.Ainsi, cette thèse démontre les effets bénéfiques de l’administration prophylactique des Lactobacillus par voie respiratoire sur la pneumonie aigue à PA. Ce résultat serait lié à l’immunomodulation de ces bactéries.Enfin, les propriétés contre l’élastase et la pyocyanine de ces souches in vitro ne présagent pas de leur activité in vivo, avec le mélange L. paracasei 9N, L. brevis 24C et L. salivarius 20C présentant seulement in vivo la meilleure activité (Brevet BIO17555)<br>The alarming increase in antibiotic resistance of Pseudomonas aeruginosa (PA) requires studying alternative therapies, such as Lactobacillus. In this thesis, several studies were carried out:1) In a murine model of acute PA pneumonia, we have demonstrated the beneficial effect of intratracheal administration of a mixture of three Lactobacillus strains from milk or the oral cavity of healthy patients.2) We then prospectively studied the Lactobacillus population in sputum of patients with cystic fibrosis (CF). The average prevalence of carry was 61%.3) Lactobacillus from the respiratory ecosystem of CF patients were used to establish two mixtures of 3 Lactobacillus strains. Strains were selected for their antielastolytic and anti-pyocyanin effects in vitro.Intranasal administration of these mixtures to C57Bl / 6 mice, 18h prior to PAO1 infection significantly improves 7-day survival and pulmonary clearance of PAO1 24h postinfection.A significant decrease in lung neutrophil recruitment and pro-inflammatory cytokines was observed, while the production of IL-10 increased.This thesis demonstrates the beneficial effects of prophylactic respiratory administration of Lactobacillus on acute PA pneumonia. Anti-PA properties in vitro are not an indication of the in vivo activity. The mixture containing L. paracasei 9N, L. brevis 24C, and L. salivarius 20C show the best anti-PA activity (Patent BIO17555). This result is likely related to an immunomodulating effect of these bacteria

Given that Streptococcus pneumoniae can cause life-threatening pulmonary and systemic infection, an apparent paradox is that the bacterium resides, usually harmlessly, in the nasopharynx of many people. Humoral immunity is thought to be the primary defense against serious pneumococcal infection, and we hypothesized that nasopharyngeal colonization of mice results in the generation of an antibody response that provides long-term protection against lung infection. We found that survival of of C57L/6 mice after intranasal inoculation with wild-type serotype 4 strain TIGR4 pneumococci required B cells but not T cells, suggesting that nasopharyngeal colonization elicited a protective humoral immune response. In fact, intranasal inoculation resulted in detectable pneumococcal-specific antibody responses, and protected mice against a subsequent high-dose S. pneumoniae pulmonary challenge. B cells were required for this response, and transfer of immune sera from i.n. colonized mice, or monoclonal antibodies against phosphorylcholine, a common surface antigen of S. pneumoniae, was sufficient to confer protection. IgA, which is thought to participate in mucosal immunity, contributed to but was not absolutely required for protection from pulmonary challenge. Protection induced by i.n. colonization lasted at least ten weeks. Although it was partially dependent on T cells, depletion of CD4+ T cells at the time of challenge did not alter protection, suggesting that T cells did not provide essential help in activation of conventional memory cells. Peritoneal B1b cells and radiation-resistant, long-lived antibody secreting cells have previously been shown to secrete anti-pneumococcal antibodies and mediate protection against systemic infection following immunization with killed bacteria or capsular polysaccharide [1, 2]. We found that peritoneal cells were not sufficient for colonization-induced protection, but sub-lethally irradiated mice largely survived pulmonary challenge. Thus, our results are consistent with the hypothesis that nasopharyngeal colonization, a common occurrence in humans, is capable of eliciting extended protection against invasive pneumococcal disease by generating long-lived antibody-secreting cells.

A large body of evidence indicates that dopamine (DA) neurotransmission regulates approach toward rewards and reward-related cues. The best-cited hypothesis proposes that DA accomplishes this by mediating the pleasurable effects of a variety of natural and drug rewards. This "anhedonia hypothesis" has received support from some pre-clinical models of reward and a few drug challenge studies in humans. However, direct assessment of DA's role in mood and other subjective states in healthy humans has been largely limited to the use of psychostimulant drugs, which elevate brain levels of multiple neurotransmitters in addition to DA. This thesis is comprised of one study which examined the effect of more selectively elevated DA neurotransmission, as produced by administration of the immediate DA precursor, L-DOPA, in healthy human volunteers. L-DOPA failed to alter mood and other subjective states. These results add to the evidence that DA neurotransmission does not directly influence mood in healthy humans.<br>La contribution précise de récompense et de motivation de la neurotransmission de la dopamine (DA) n'est pas entièrement comprise. La meilleure hypothèse citée propose que la DA forme un trait d'union des effets agréables d'une variété de récompenses naturelles et narcotiques. Cette hypothèse « anhédoniste » a reçu l'appui de quelques modèles précliniques de récompense et de quelques études chez l'humain mettant la drogue en question. Cependant, l'évaluation directe du rôle de la DA sur la disposition et autres états subjectifs chez l'humain en santé a été principalement limitée à l'utilisation de drogues psychostimulantes, ce qui élève le niveau de neurotransmetteurs multiples en plus de la DA dans le cerveau. La présente thèse comprend une étude qui examine l'effet d'une neurotransmission sélectivement plus élevée de la DA, produite par l'administration du précurseur immédiat de la DA, L-DOPA, chez des volontaires en santé. L-DOPA n'a modifié ni disposition ni autres états subjectifs. Ces résultats s'ajoutent à l'évidence la neurotransmission de DA n'influence pas directement la disposition chez l'humain en santé.

Le virus de la grippe infecte les muqueuses du tractus respiratoire. Un vaccin intranasal induit une réponse immunitaire proche de celle faisant suite à une infection naturelle en bloquant le virus directement sur le site de l'infection et permet une vaccination sans aiguille. Par ailleurs, les vaccins à base de pseudo-particules virales ou Virus-like Particles (VLP) produites sur cellules représentent une alternative intéressante au vaccin classique produit sur oeufs. Les VLP sont des particules non-réplicatives qui ressemblent au virus et qui peuvent être immunogènes même sans adjuvant, en particulier par voie intranasale. Au cours de ma thèse, une plateforme de production de VLP grippales composées d'hémagglutinine, de neuraminidase et de protéine de matrice M1 a été développée par transfection transitoire des cellules de mammifères. Des immunisations de souris BALB/c ont montré que les VLP de type A et B, purifiées et caractérisées, étaient immunogènes à de faibles doses par voie intramusculaire. L'administration par voie intranasale de VLP avec la sous-unité B de la toxine cholérique, comme adjuvant muqueux, a permis d'obtenir des taux d'anticorps sériques comparables à ceux obtenus par immunisation en intramusculaire mais également une forte réponse IgA au niveau des muqueuses. Par ailleurs, le rendement des VLP s'est révélé souche-dépendant et lié aux protéines HA et NA à la surface de la particule. Pour contourner ce problème, un vaccin quadrivalent composé de deux VLP bivalentes exprimant chacune deux HA et NA différentes à la surface a été produit montrant ainsi la flexibilité de cette plateforme<br>The influenza virus infects the mucous membranes of the respiratory tract. An intranasal vaccine induces an immune response close to the one induced by the natural infection by blocking the virus directly at the site of infection and allows needle-free vaccination. In addition, vaccines based on Virus-like Particles (VLP) produced in cells represent an interesting alternative to the traditional egg-based vaccine. VLPs are non-replicative particles that mimic the virus. Studies on influenza VLPs have shown protection by the intranasal route without adding an adjuvant. During my thesis, a platform for the production of influenza VLPs composed of the hemagglutinin, the neuraminidase and the M1 matrix proteins was developed by transient transfection of mammalian cells. Immunizations of BALB/c mice showed that the purified and characterized type A and B VLPs were immunogenic at low doses by the intramuscular route. The intranasal administration of VLPs with the B subunit of cholera toxin as a mucosal adjuvant resulted in serum antibody levels comparable to those obtained by intramuscular immunization but also a strong IgA response in the mucosal secretions. In addition, VLP yield was found to be strain-dependent and linked to the HA and NA proteins on the surface of the particle. To overcome this problem, a quadrivalent vaccine based on two bivalent VLPs each expressing two different HAs and NAs at the surface was produced, demonstrating the flexibility of this platform

Orientador: Marlus Chorilli<br>Resumo: O diabetes mellitus (DM) é uma síndrome metabólica caracterizada por deficiência na produção/secreção pancreática de insulina e/ou resistência à ação do hormônio nos tecidos alvo, resultando em hiperglicemia. Diversas pesquisas têm desencadeado o desenvolvimento de novos sistemas de administração de insulina que possibilitem a utilização de vias alternativas à parenteral, com destaque à administração de insulina por via nasal. Esta via tem-se mostrado promissora, pois pode promover uma rápida absorção do fármaco e aumentar a sua biodisponibilidade. Entretanto, existem mecanismos de depuração mucociliar que limitam a administração de fármacos, além da baixa permeabilidade do epitélio nasal, o qual dificulta a absorção de fármacos com alto massa molar. Uma estratégia para vencer tais barreiras é a utilização de sistemas nanoestruturados (lipossomas), pois são amplamente utilizados para o aperfeiçoamento da potencialização da ação terapêutica de fármacos. Além disso estes lipossomas foram funcionalizados com peptídeos de transdução de membrana (CPPs), tais como os peptídeos TAT e Penetratin (PNT), que atuam como promotores da penetração e absorção do fármaco, com posterior dispersão em hidrogel de hidroxietilcelulose. O objetivo deste trabalho foi desenvolver e caracterizar lipossomas contendo solução de insulina, funcionalizados com CPPs (TAT e PNT) e dispersos em hidrogel, avaliar o potencial pela via nasal, in vivo, para a melhora dos níveis séricos e efeito hipoglicemiante d... (Resumo completo, clicar acesso eletrônico abaixo)<br>Doutor

This research project investigates the long-term consequences of perinatal exposure to high-fat (HF) on the mesocorticolimbic dopamine (DA) system. Adult offspring of mothers fed a HF diet (30% fat, compared to 5% in control mothers (C)) during the last week of gestation and throughout lactation displayed decreased locomotion in response to an acute amphetamine challenge and decreased behavioral sensitization to repeated amphetamine compared to C animals. These behavioral effects were accompanied by small increases in tyrosine hydroxylase expression in the ventral tegmental area and significant increases in DA and DOPAC content in the NAc, suggesting an elevated DA tone in this target field. In the NAc, there were no significant changes in D1, D2 receptors or DA transporter (DAT) levels between diet groups. The behavioural and biochemical data were collected in adulthood, long after the termination of the diet suggesting that a HF perinatal diet is inducing permanent changes within the DA system and might contribute to the development of metabolic disturbances.

Stress has been reported to activate the locus coeruleus (LC)-noradrenergic system. In this study, corticosterone (CORT) was orally administrated to rats for 21 days to mimic stress status. In situ hybridization measurements showed that CORT ingestion significantly increased mRNA levels of norepinephrine transporter (NET) and dopamine β-hydroxylase (DBH) in the LC region. Immunofluorescence staining and western blotting revealed that CORT treatment also increased protein levels of NET and DBH in the LC, as well as NET protein levels in the hippocampus, the frontal cortex and the amygdala. However, CORT-induced increase in DBH protein levels only appeared in the hippocampus and the amygdala. Elevated NET and DBH expression in most of these areas (except for NET protein levels in the LC) was abolished by simultaneous treatment with combination of corticosteroid receptor antagonist mifepristone and spironolactone (s.c. for 21 days). Also, treatment with mifepristone alone prevented CORT-induced increases of NET expression and DBH protein levels in the LC. In addition, behavioral tasks showed that CORT ingestion facilitated escape in avoidance trials using an elevated T-maze, but interestingly, there was no significant effect on the escape trial. Corticosteroid receptor antagonists failed to counteract this response in CORT-treated rats. In the open-field task, CORT treatment resulted in less activity in a defined central zone compared to controls and corticosteroid receptor antagonist treatment alleviated this increase. In conclusion, this study demonstrates that chronic exposure to CORT results in a phenotype that mimics stress-induced alteration of noradrenergic phenotypes, but the effects on behavior are task dependent. As the sucrose consumption test strongly suggests CORT ingestion-induced depression-like behavior, further elucidation of underlying mechanisms may improve our understanding of the correlation between stress and the development of depression.

Caffeine is the most consumed psychoactive substance in the world, and most caffeine consumption in coffee and energy drinks is intended to produce a psychoactive effect. However, caffeine is not a primary reinforcer in preclinical paradigms – non-human species do not reliably take the drug to produce a psychoactive effect. However, caffeine is a ‘reinforcement enhancer’ in preclinical models; the effects of caffeine increase the motivation to obtain other non-drug reinforcers. The overall goal of this project was to determine if these reinforcement enhancing effects of caffeine could promote caffeine self-administration and to subsequently investigate the behavioral and neurochemical underpinnings of this effect. We hypothesized reliable caffeine self-administration would occur by adventitious pairing of caffeine with saccharin, a primary reinforcer. Second, we hypothesized that caffeine enhances reinforcement by increasing the salience of incentive stimuli, which are stimuli that come to evoke approach behaviors through associative learning (e.g., Pavlovian conditioning). Finally, incentive salience is moderated by dopamine release in the nucleus accumbens (NAc), an area highly involved in reward-learning and substance dependence. Therefore, we hypothesized that if caffeine enhanced control of approach behavior by incentives, then it would increase the ability of incentive stimuli to evoke dopamine in the NAc. These studies show that intravenous delivery of caffeine with oral saccharin increases operant relative to control groups responding for intravenous caffeine or oral saccharin. The effect was also dose-dependent, confirming that the psychoactive effects of caffeine increased behavior. We also extended this effect to an oral model of caffeine self-administration, which included a simple sweetener (saccharin) or a complex oral vehicle (saccharin with decaffeinated coffee) to mask the bitter taste of caffeine. Presenting caffeine with oral saccharin promoted self-administration, relative to saccharin alone and did not depend on the nature of the complexity of the vehicle. Caffeine also dose-dependently increased approach to an incentive stimulus and this effect was associated with increased extracellular dopamine in the NAc. These findings suggest caffeine enhances incentive motivation and that this effect may result from increases in CS-evoked striatal dopamine.

Caffeine is a methylxanthine that acts as a nonselective adenosine antagonist, binding to adenosine A1 and A2A receptor subtypes. This substance is highly present in different beverages like coffee, tea and in energy drinks. Epidemiology studies have shown a positive correlation between the consumption of energy drinks and that of ethanol. Combination of both substances could affect alcohol consumption patterns and can modulate effects like sedation or motor incoordination. Adenosine acts on A1 and A2A receptors, which are located in brain areas involved in motivational processes. Adenosine also interacts with other neurotransmitters such as dopamine. The present research also assessed the impact of caffeine on behavioral procedures that induce individual differences in effort expenditure for food seeking behavior. Effort-related dysfunctions are seen in many psychopathologies, thus the study of individual differences could be useful to optimize treatments and to look for alternative treatments based on the adenosine system.<br>La cafeína es una metilxantina que actúa como un antagonista no selectivo de los receptores de adenosina A1 y A2A. Esta sustancia está presente en bebidas como café, té y bebidas energéticas. Los estudios epidemiológicos han mostrado una correlación positiva entre el consumo de bebidas energéticas y de etanol. La combinación de ambas sustancias podría afectar los patrones de consumo de alcohol y modular efectos como la sedación o la incoordinación motora. Los receptores A1 y A2A, se localizan en áreas cerebrales involucradas en procesos motivacionales en las que interactúa con la dopamina. También se evaluó el impacto de la cafeína en procedimientos conductuales que inducen diferencias individuales en la predisposición a realizar esfuerzo. Las disfunciones relacionadas con el esfuerzo se observan en muchas psicopatologías, por lo que el estudio de las diferencias individuales podría ser útil para optimizar los tratamientos y buscar otros alternativos basados en el sistema adenosinérgico.

Background: A single infusion of ketamine has rapid antidepressant properties, although the drawback is a lack of sustained effect. A previous study showed a rapid enhancement (within 2 hours) in ventral tegmental area (VTA) dopamine (DA) neuron population and locus coeruleus (LC) norepinephrine (NE) firing and bursting activity following a single ketamine administration. The current study investigated whether these changes are present 24 hours after a single administration and if they are maintained with repeated administration. Additionally, we examined dorsal raphe nucleus (DRN) serotonin (5-HT) neurons to assess the effects of single and repeated ketamine administration on these neurons. Methods: Ketamine (10 mg/kg, i.p.) was administered to male Sprague Dawley rats once or repeatedly (3 times/week) for 2 weeks. After single and repeated administration of ketamine, electrophysiological recordings were done in the VTA, LC and DRN in anesthetized rats, 24 hrs, 3 or 7 days post-administration. Spike frequency, bursting, and for VTA neurons, spontaneously active neurons/trajectory were assessed. Results: In the VTA, LC and DRN, 24 hrs after ketamine was injected acutely there was no significant difference between controls and treated animals in all parameters assessed. However, after repeated administration, there was an increase in bursting and number of spontaneously discharging neurons per tract of VTA DA neurons as well as an increase in frequency of discharge of LC NE neurons. While the increased number of spontaneously discharging neurons per tract had dissipated after 3 days, the enhanced bursting was still present but dissipated after 7 days. As for LC NE neurons, the increased frequency of discharge was no longer present after 3 days. No significant differences in the firing of DRN 5-HT neurons were observed between controls and treated animals even after ketamine was administered repeatedly. Conclusion: These results indicate that repeated but not acute administration of ketamine maintained the increase in population activity of DA neurons and firing activity of NE neurons.

Background: Dopamine levels has been implicated in obesity, feeding behaviour, and hedonic control of appetite like olfactory cues and food palatability. Methylphenidate (MPH) is a dopamine reuptake inhibitor that increases brain dopamine levels and has been shown to reduce appetite and promote weight loss in patients with attention deficit hyperactivity disorder (ADHD). As such, the objectives of this study were to test the possible effect of MPH on appetite, olfaction, and food palatability as well as its effects on energy intake and body weight of healthy individuals with obesity. Methods: In a randomized, double-blind study, 12 participants (age 28.9±6.7 yrs) (BMI 36.1±4.5 kg/m2) were assigned to receive MPH (0.5mg/kg) (n=5) or placebo (n=7) twice daily for two months. Appetite and palatability (Visual Analog Scale (VAS)), odour threshold (Sniffin’ Sticks®), in-lab energy intake (ad libitum buffet), free-living energy intake (3-day food boxes) and body weight (DEXA scan) were measured at baseline (day 1) and final visit (day 60). Results: MPH intake caused significantly greater suppression of appetite sensations (desire to eat (p=0.001), hunger (p=0.008), and prospective food consumption (p=0.003)) and increase in fullness (p=0.028) over time compared to placebo. There was a significant increase in odour threshold scores in the MPH group (6.3±1.4 vs. 9.4±2.1) compared to placebo (7.9±2.3 vs. 7.8±1.9) (p=0.029). Both placebo and MPH groups showed decreases in their energy intake (p=0.021) and body weight (p=0.005) over time but with large effect sizes favouring greater reduction in the MPH group compared to placebo. Conclusions: Compared to placebo, MPH intake over 60 days suppressed appetite and improved olfactory sensitivity in individuals with obesity. These data provide novel findings into the possible efficacy of MPH to favourably impact appetite and therefore promoting weight loss in individuals living with obesity.

Binge alcohol consumption is a rising concern in the United States, especially among adolescents as during this developmental period alcohol use is usually initiated and has been shown to cause detrimental effects on brain structure and function. These findings have been established through the use of binge models in animals, where animals are repeatedly administered high doses of ethanol typically over a period of three or four days. While such work has examined the effects of a four-day and repeated three-day binge, there has been almost no work conducted aimed at investigating the long-term behavioral and neurochemical and/or functional consequences of repeated binge pattern administration during adolescence relative to adulthood on later ethanol-induced behavior and neurochemistry in adulthood. The present set of experiments aimed to examine the dose-response and age-related differences induced by repeated binge pattern ethanol administration during adolescence or adulthood on voluntary ethanol consumption (Aim 1), changes in ethanol metabolism following ethanol pretreatment (Aim 2) and mesolimbic dopamine functionality (Aim 3) in adulthood. In both experiments, adolescent and adult male rats were intragastrically administered ethanol (0.5, 1.0 or 2.0 g/kg/ig) or isovolumetric water on postnatal days (PND) 28-31, PND 35-38 and PND 42-45 for adolescent rats and PND 60-64, PND 67-70 and PND 74-77 for adult rats. In both experiments all rats underwent fourteen days of abstinence (PND 46-59 or PND 78-91, respectively). Subsequently, in Experiment 1, all rats underwent voluntary ethanol consumption procedures, in which animals were exposed to 10% ethanol combined with decreasing saccharin concentrations across days from PND 60-82 for adolescent-exposed rats and PND 92-114 for adult-exposed rats. Finally, on PND 83 and PND 115, respectively, all animals were challenged with 2.0 g/kg ethanol and trunk blood samples were collected at 60 and 240 minutes post-injection. Results indicate there was a significant increase in voluntary ethanol intake in adolescent ethanol-exposed rats pretreated with 2.0 g/kg relative to their adult ethanol-pretreated counterparts. Faster ethanol metabolism was observed in adolescent rats pretreated with 2.0 g/kg during adolescence relative to adolescent-exposed rats pretreated with 0.5 g/kg and adults pretreated with 2.0 g/kg. For Experiment 2, all rats underwent surgery (PND 60 for adolescent-exposed and PND 92 for adult-exposed rats). From PND 61-64 for adolescent-exposed and PND 93-96 for adult exposed rats, all animals underwent recovery from surgery. Finally, all rats underwent in vivo microdialysis on PND 65 for adolescent-exposed and PND 97 for adult-exposed rats, with K+ (100 mM) infused into the ventral tegmental area and accumbal dopamine overflow assessed in the nucleus accumbens septi. The results from Experiment 2 indicate lasting changes in mesolimbic dopamine functionality with a trend for decreased potassium-stimulated dopamine overflow in the nucleus accumbens septi in adolescent-ethanol pretreated rats and a trend for increased potassium-stimulated dopamine overflow in adult ethanol-pretreated rats. The results from the present set of experiments show the dose-dependent impact of binge-pattern ethanol exposure during adolescence on subsequent ethanol consumption and ethanol metabolism in adulthood. These findings indeed determine adolescence as a period of vulnerability to the long-term changes in ethanol consumption relative to similarly-exposed adult male rats. Importantly, the results of Experiment 2 indicate an alteration in the functionality of the mesolimbic pathway in adulthood following adolescent binge pattern ethanol exposure, which demonstrates a long-term depression in mesolimbic dopamine functionality following adolescent binge pattern ethanol exposure.

Schizophrenia affects about 1% of the population. A hallmark of the disorder is increased dopamine D2 receptor sensitivity in the brain. Studies have shown that schizophrenics smoke cigarettes at approximately 4 times the rate of the general population. It has been suggested that nicotine use is a form of self-medication for symptoms in schizophrenia. Smoking behaviors typically begin in adolescence. We assessed effects of nicotine on behavior and brain plasticity in an adolescent rodent model of schizophrenia with the goal of identifying targets for smoking cessation. Methods: Rats were neonatally treated with quinpirole (a D2/D3 agonist) or saline and sensitized to 0.3, 0.5, or 0.7 mg/kg (free base) nicotine or saline, every other day for 9 days, and locomotor activity was recorded. After behavioral testing, animals demonstrating sensitization to 0.5mg/kg nicotine were surgically implanted with a guide cannula, aimed at the nucleus accumbens core. After recovery, animals underwent microdialysis and in vivo samples were collected every 20 minutes for 300 minutes. Postmortem brains from animals exposed to 0.5mg/kg nicotine or saline were dissected and the nucleus accumbens and dorsal striatum were analyzed for brain-derived neurotrophic factor (BDNF), phosphorylated cAMP response element binding protein (pCREB), and glial-cell derived neurotrophic factor (GDNF), all proteins involved in neuronal plasticity. Results: Animals neonatally treated with quinpirole and administered nicotine showed robust increases in locomotor sensitization and a 400% increase in dopamine overflow from the accumbens core, which was greater than all other groups. Nicotine administration led to increased accumbal BDNF levels, which was enhanced by neonatal quinpirole pretreatment. GDNF levels were also increased in control animals given nicotine, which was attenuated to control levels by neonatal quinpirole. Finally, pCREB levels were robustly increased in animals neonatally treated with quinpirole, an effect that was partially attenuated by adolescent nicotine treatment. These data on pCREB suggest a possible biological marker of anhedonia. In conclusion, it is apparent that nicotine results in a robust increase in behavioral activity and changes in neural proteins of brain plasticity that may serve as possible pharmaceutical targets for smoking cessation in schizophrenia.

Pharmaceutical Sciences<br>Ph.D.<br>Illicit drug use has been a growing concern over the past few decades. The rise in use of illegal drugs drove the government and law enforcement to aggressively tackle this problem and crackdown on the illicit use of drugs. However, this sparked a further interest in ‘legal highs.’ Before 2011, among the newly popular ‘legal highs’ was ‘Bath Salts.’ Cathinone is a monoamine alkaloid and the active ingredient found in the leaves of the khat plant. The psychoactive form of bath salts may contain a mixture of synthesized cathinones, including, 4-methyl-N-methcathinone (mephedrone), 3,4-methylenedioxy-N- methylcathinone (methylone) and methylenedioxypyrovalerone (MDPV). These three are commonly found in bath salts. One of the major psychoactive ingredients in bath salts is mephedrone. Mephedrone grew in popularity due to its low price, accessibility, and the shortage of MDMA, thus making mephedrone the prime drug to sell as a ‘legal high’ up until 2011 when it became banned in the United S<br>Temple University--Theses

In patients with Parkinson's disease (PD), the associated pathology follows a characteristic pattern involving inter alia the enteric nervous system (ENS), the dorsal motor nucleus of the vagus (DMV), the intermediolateral nucleus of the spinal cord and the substantia nigra, providing the basis for the neuropathological staging of the disease. Here we report that intragastrically administered rotenone, a commonly used pesticide that inhibits Complex I of the mitochondrial respiratory chain, is able to reproduce PD pathological staging as found in patients. Our results show that low doses of chronically and intragastrically administered rotenone induce alpha-synuclein accumulation in all the above-mentioned nervous system structures of wild-type mice. Moreover, we also observed inflammation and alpha-synuclein phosphorylation in the ENS and DMV. HPLC analysis showed no rotenone levels in the systemic blood or the central nervous system (detection limit [rotenone]<20 nM) and mitochondrial Complex I measurements showed no systemic Complex I inhibition after 1.5 months of treatment. These alterations are sequential, appearing only in synaptically connected nervous structures, treatment time-dependent and accompanied by inflammatory signs and motor dysfunctions. These results strongly suggest that the local effect of pesticides on the ENS might be sufficient to induce PD-like progression and to reproduce the neuroanatomical and neurochemical features of PD staging. It provides new insight into how environmental factors could trigger PD and suggests a transsynaptic mechanism by which PD might spread throughout the central nervous system.

L'administration nasale a un grand avantage pour stimuler l'immunité protectrice locale et systémique. Cependant, des systèmes d'administration et des adjuvants sont souvent nécessaires pour améliorer l'efficacité du vaccin intranasal. Nous avons appliqué la technologie des nanoparticules en tant que système universel de délivrance de vaccins contre la grippe dans le projet européen FP7 appelé UniVacFlu.Nous avons évalué différentes nanoparticules (NP) pour rechercher le meilleur nanovecteur. Pour cela, nous avons comparé 5 types de nanoparticules avec différentes charges de surface (anioniques ou cationiques) et diverses compositions internes comme vecteurs potentiels: des liposomes cationiques ou anioniques, des NP de PLGA cationique ou anionique (poly acide lactique co-glycolique) et une NP cationique composée de maltodextrine fonctionnalisée par un agent cationisant avec un coeur de lipides anioniques (NPL). Nous avons d'abord quantifié leur temps de résidence nasale après l'administration nasale chez la souris en utilisant l'imagerie in vivo et les NPL ont montré le plus long temps de résidence. L'endocytose in vitro sur des cellules muqueuses (cellules épithéliales des voies respiratoires, macrophages et cellules dendritiques) en utilisant des nanoparticules marquées a été réalisée par cytométrie de flux et microscopie confocale. Parmi les 5 nanoparticules, les NPL ont été majoritairement captées par 3 lignées cellulaires différentes représentatives d’un épithélium respiratoire et les mécanismes d'endocytose ont été caractérisés. Afin d’évaluer le meilleur vecteur en tant que véhicules, le chargement d'antigènes et la délivrance intracellulaire ont été évalués dans des cellules de la muqueuse des voies respiratoires (cellules épithéliales des voies aériennes, macrophages et cellules dendritiques) par cytométrie de flux. Nous montrons que les NPL sont les meilleurs candidats capables de délivrer la plus grande quantité de protéines dans les cellules. Pris ensemble, notre étude a révélé que parmi 5 nanoparticules, la NPL était le meilleur nanovecteur en termes de temps de résidence nasale, d'endocytose par les cellules et de délivrance de protéines dans l'épithélium des voies respiratoires. Les NPL ont donc été sélectionnées comme nanovecteurs pour le projet UniVac Flu.Les antigènes de la grippe CTA1-3M2e-DD et HA ont été formulés avec les NPL. Le CTA1-3M2e-DD est un antigène chimérique adjuvanté et ciblé. Il est composé de la sous-unité A1 de la toxine du choléra et un épitope conservé du virus grippal A (M2e), ainsi que le dimère de l'analogue synthétique de la protéine A de Staphylococcus aureus (DD) utilisé comme agent de ciblage des lyphocytes B. Pour améliorer l'effet antigénique, l’HA recombinant de H1N1 a été combinée avec CTA1-3M2e-DD. Ces formulations ont été évaluées chez la souris par le consortium UniVacFlu. Les résultats ont montré que CTA1-3M2e-DD et HA chargé dans les NPL formeraient un vaccin intranasal prometteur contre la grippe. Ce travail de thèse montre que les NPL sont des nanovecteurs d’intérêt pour le vaccin nasal<br>Nasal administration has great advantage for stimulating the immune system, such as stimulating local and systemic protective immunity. However, delivery systems and adjuvants are often necessary to improve the efficacy of the intranasal vaccine. We applied nanoparticle technology to deliver a universal influenza vaccine via the nasal route in a European FP7 project called UniVacFlu.We evaluated different nanoparticles to search the best nanocarrier for an intranasal vaccine. Here we compared 5 types of nanoparticles with different surface charges (anionic or cationic) and various inner compositions as potential vectors: cationic and anionic liposomes, cationic and anionic PLGA (Poly Lactic co-Glycolic Acid) nanoparticles and zwitterionic maltodextrin nanoparticles (cationic surface with an anionic lipid core: NPL). We first quantified their nasal residence time after nasal administration in mice using in vivo live imaging and NPL showed the longest residence time. In vitro endocytosis on mucosal cells (airway epithelial cells, macrophages and dendritic cells) using labeled nanoparticles were performed by flow cytometry and confocal microscopy. Among the 5 nanoparticles, NPL were taken up to the greatest extent by the 3 different cell lines and the endocytosis mechanisms of NPL were characterized. In order to compare different nanoparticles as vaccine carriers, antigen loading and cell delivery were evaluated. In this study, we compared the loading and delivery of labeling ovalbumin with airway mucosa cells (airway epithelial cells, macrophages and dendritic cells) by flow cytometry. Our data showed that NPL were the best candidate that can payload with highest amount of protein and eventually the most efficient cellular protein delivery capacity. Taken together, our study revealed that among 5 nanoparticles, NPL were the best nanocarrier that own longer nasal residence time, efficiently uptake and deliver protein into airway epithelium. NPL were then selected as nanocarrier for the UniVac Flu project.The flu antigens CTA1-3M2e-DD and HA were formulated with NPL. The CTA1-3M2e-DD is an adjuvanted antigen composed of the A1 subunit of cholera toxin and a conserved epitope of influenza A virus (M2e), as well as the dimer of the synthetic analogue of Staphylococcus aureus protein A (DD) used to target B cells. To improve antigenic effect, recombinant HA from H1N1 was combined with CTA1-3M2e-DD. These formulations were evaluated in mice by the UniVacFlu consortium. We observed that CTA1-3M2e-DD and HA loaded into NPL could be a promising universal intranasal influenza vaccine

Cocaine dependence is a major health concern worldwide, but despite this high rate of abuse there are currently no approved therapies for cocaine dependence. Replacement pharmacotherapies are one possible approach for treating cocaine dependence, and identification of such therapeutics for cocaine abuse is the long-term goal of this research. Cocaine binds to, and inhibits uptake at the dopamine (DAT), serotonergic (SERT) and noradrenaline (NET) uptake transporters, but studies have shown that cocaine produces its strong behavioural and positive reinforcing effects through inhibition of the DAT. To this end a great number of diverse, non-selective DAT-inhibiting compounds have been investigated as potential cocaine replacement therapies. It was the initial objective of this research to determine whether the behavioral profile of a novel, selective DAT inhibitor, D-84, fit with that thought for an ideal cocaine replacement therapy. Results indicated that D-84 stimulated locomotor activity, incompletely generalized to the cocaine cue in discrimination tests, attenuated cocaine-self-administration and was self-administered. These observations provide a profile consistent, although perhaps not ideal, with one possible treatment strategy for cocaine dependence. Although it is well established that cocaine predominantly produces its abuse-related effects through inhibition of the DAT, recent evidence suggests that inhibition at the SERT may have modulating effects on the pharmacology of cocaine-like compounds. The second part of this dissertation investigated what effects that increasing SERT inhibition had on the cocaine-like behavioural effects of DAT inhibitors, as a method of determining the fruitfulness of incorporating this feature into future drug candidates to improve them. RTI-55 (DAT Ki 2.7 nM SERT Ki 3 nM) and GBR-12909 (DAT Ki 4.3 nM SERT Ki 73 nM) were selected based on their high and intermediate SERT inhibitory effects, respectively. They were compared in behavioural studies with D-84, which is considered to be a selective DAT inhibitor. The results indicated that although increasing SERT inhibition attenuated locomotor activity effects, it had less effect on cocaine-like discriminative stimulus and reinforcing effects, at least with the doses tested

In patients with Parkinson's disease (PD), the associated pathology follows a characteristic pattern involving inter alia the enteric nervous system (ENS), the dorsal motor nucleus of the vagus (DMV), the intermediolateral nucleus of the spinal cord and the substantia nigra, providing the basis for the neuropathological staging of the disease. Here we report that intragastrically administered rotenone, a commonly used pesticide that inhibits Complex I of the mitochondrial respiratory chain, is able to reproduce PD pathological staging as found in patients. Our results show that low doses of chronically and intragastrically administered rotenone induce alpha-synuclein accumulation in all the above-mentioned nervous system structures of wild-type mice. Moreover, we also observed inflammation and alpha-synuclein phosphorylation in the ENS and DMV. HPLC analysis showed no rotenone levels in the systemic blood or the central nervous system (detection limit [rotenone]<20 nM) and mitochondrial Complex I measurements showed no systemic Complex I inhibition after 1.5 months of treatment. These alterations are sequential, appearing only in synaptically connected nervous structures, treatment time-dependent and accompanied by inflammatory signs and motor dysfunctions. These results strongly suggest that the local effect of pesticides on the ENS might be sufficient to induce PD-like progression and to reproduce the neuroanatomical and neurochemical features of PD staging. It provides new insight into how environmental factors could trigger PD and suggests a transsynaptic mechanism by which PD might spread throughout the central nervous system.

Preference for palatable and energy-dense foods may be a risk factor for body weight gain and has both genetic and environmental components. Once obesity develops in an individual, weight loss is difficult to achieve. Indeed, obesity is often characterized by repeated attempts to reduce the overconsumption of energy-dense foods, followed by food craving and relapse to overconsumption. Relapse and loss of control over intake are observed also in drug addicts, and it has been shown that obesity and drug addiction not only share behavioural features but also neural circuitry, e.g. the mesolimbic dopamine pathway. In this thesis, we sought to investigate the mechanisms related to food preferences and craving using animal models previously used in addiction research. The risk of gaining weight may implicate behavioural traits and emotional states. We showed in rats that a risk-taking behavioural profile was associated both with increased preference for a high-fat (HF) diet and with increased motivational response to a palatable high-sucrose (HS) diet. Hypothalamic urocortin 2 expression was associated with the preference for the HF diet. We also tested the hypothesis that consumption of HS and HF diets separately or provided simultaneously (HFHS) affect anxiety-like behaviour and locomotion. Furthermore, we showed that withdrawal from HFHS food affects diet-induced obesity-prone (OP) and obesity-resistant (OR) animals differently. OP animals had increased motivation (craving) for HS food pellets as measured by the operant self-administration technique during withdrawal. Dopamine receptor expression in the striatum differed between OP and OR animals both at access to HFHS and during withdrawal. This strongly implicates dopaminergic signaling in the OP phenotype. In humans, food preferences may be monitored using questionnaires. We analyzed food preference data from parents of preschool children, and identified an inverse association of parental preference for high-fat high-protein food and overweight in children. In conclusion, we have employed animal models previously used in the addiction field to identify molecular mechanisms related both to food preference and vulnerability to obesity, and to food craving associated with withdrawal from palatable food. These findings add to our current understanding of obesity.

A key issue in neuroscience is to determine the connection between neuronal circuits and behaviour. In the adult brain, all neuronal circuits include a glutamatergic component. Three proteins designated Vesicular glutamate transporter 1-3 (VGLUT1-3) possess the capability of packaging glutamate into presynaptic vesicles for release of glutamate at the nerve terminal. The present study aimed at determining the role of VGLUT2 in neuronal circuits of higher brain function, emotion, and reward-pocessing. A conditional knockout (cKO) strategy was utilised, and three different mouse lines were produced to delete VGLUT2 in specific neuronal circuits in a temporally and spatially controlled manner. First, we produced a cKO mouse in which Vglut2 was deleted in specific subpopulations of the cortex, amygdala and hippocampus from preadolescence. This resulted in blunted aspects in cognitive, emotional and social behaviour in a schizophrenia-related phenotype. Furthermore, we showed a downstream effect of the targeted deletion on the dopaminergic system. In a subsequent analysis of the same cKO mice, we showed that female cKO mice were more affected their male counterparts, and we also found that female schizophrenia patients, but not male patients, had increased Vglut2 levels in the cortex.  Second, we produced and analysed cKO mice in which Vglut2 was deleted in the cortex, amygdala and hippocampus already from midgestation, and could show that this deletion affected emotional, but not cognitive, function. Third, we addressed the role of VGLUT2 in midbrain dopamine neurons by targeting Vglut2 specifically in these neurons. These cKO mice showed a blunted activational response to the psychostimulant amphetamine and increased operant self-administration of both sugar and cocaine reinforcers. Further, the cKO mice displayed strongly enhanced cocaine-seeking in response to cocaine-associated cues, a behaviour of relevance for addiction in humans. In summary, this thesis work has addressed the role of the presynaptic glutamatergic neuron in different neuronal circuits and shown that the temporal and spatial distribution of VGLUT2 is of great significance for normal brain function.

Le glioblastome (GB), tumeur primitive du cerveau, la plus agressive, et la plus fréquente chez l’adulte, présente une prolifération vasculaire importante. Des agents thérapeutiques innovants ciblant à la fois l'angiogenèse et les cellules tumorales sont recherchés, ainsi que des systèmes pour augmenter leur délivrance dans la tumeur cérébrale. Un de ces agents est le sorafénib (SFN), un inhibiteur de tyrosine kinase. Sa mauvaise solubilité aqueuse et ses effets secondaires indésirables limitent son utilisation. Le premier objectif de cette thèse était d'encapsuler cet agent dans des nanocapsules lipidiques (NCL) pour contrer ces inconvénients. Nous avons développé des NCL avec une haute efficacité d'encapsulation du SFN qui inhibaient in vitro l'angiogenèse et la viabilité de la lignée de GB humain U87MG. La délivrance intratumorale de SFN-NCL chez des souris porteuses d’une tumeur intracérébrale U87MG induit une normalisation vasculaire tumorale précoce qui pourrait améliorer l'efficacité de la chimiothérapie et de la radiothérapie. Le second objectif était de définir si la délivrance intranasale de NCL pouvait constituer une voie non-invasive alternative. Nous avons étudié via le transfert d'énergie par résonance de type Förster, le devenir des NCL chargées d’un fluorochrome à travers des monocouches de cellules Calu-3, un modèle de l'épithélium nasal. L'utilisation de NCL augmente le passage du fluorochrome à travers les cellules Calu-3, mais les particules sont rapidement dégradées après leur capture. Ces données mettent en évidence que les NCL sont appropriées pour la délivrance locale du SFN mais doivent être modifiées pour une délivrance intranasale<br>Glioblastoma (GB), the most aggressive, and the most frequent primary tumor of the brain in adults, present a prominent vascular proliferation. Innovative therapeutic agents targeting both angiogenesis and tumor cells are urgently required, along with competent systems for their delivery to the brain tumor. One such agent is sorafenib (SFN), a tyrosine kinase inhibitor. However, poor aqueoussolubility and undesirable side effects limit its clinical application. The first objective of this thesis was to encapsulate this drug inside lipid nanocapsules(LNCs) to overcome these drawbacks. We developed LNCs with a high SFN encapsulation efficiency (&gt;90%) that inhibited in vitro angiogenesis and the viability of the human U87MG GB cell line. Intratumoral delivery of SFN-LNCs in mice bearing intracerebral U87MG tumors induced early tumor vascular normalization which could be used to improve the efficacy of chemotherapy and radiotherapy in the treatment of GB. The second objective was to define whether intranasal delivery of LNCs could be an alternative non-invasive route. In this regard, we investigated through Förster resonance energy transfer, the fate of dye-loaded LNCs across Calu-3 cell monolayers, a model of the nasal mucosa. We showed that employment of LNCs dramatically increased the delivery of the dye acrossCalu-3 cell monolayer but they were rapidly degraded after their uptake. These data highlight that LNCs are suitable nanocarriers for the local delivery of SFN but must be redesigned for enhancing their nose-to-brain delivery

La modification de l'activité neuronale peut engendrer des altérations dans des circuits neuronaux. L'activation des récepteurs couplés aux protéines G (RCPG) peux participer à des mécanismes à la base du développement de maladies comme l'addiction à la cocaïne. La consommation de cocaïne conduit à une augmentation de neurotransmetteurs tels que la dopamine, l'histamine et le glutamate qui activent des RCPG dans le cerveau. La signalisation des RCPG peut se faire au travers de monomères, d'homo- ou d'hétéromères de RCPG ainsi que par des interactions protéine - protéine, permettant entre autre une régulation croisée. Nous montrons que les récepteurs de la dopamine 1 (D1R) et de l'histamine 3 (H3R) induisent une signalisation croisée dans le striatum de rat vraisemblablement par la formation d'hétérodimères. Une administration chronique de cocaïne modifie la signalisation de ces récepteurs tant que la signalisation croisée des D1R et H3R. Les H3R et les récepteurs métabotropes du glutamate 1/5 (mGlu1/5R) sont fortement exprimés dans l'hippocampe et le striatum. Des expériences de comportement suggèrent que ces récepteurs seraient susceptibles de coordonner leurs signalisations par une régulation croisée. Nos expériences d'électrophysiologie, de mesure de Ca++ intracellulaire et de transduction du signal montrent effectivement une régulation croisée des récepteurs H3R et mGlu1/5R dans le cerveau de rat. De plus, nous montrons que la consommation chronique de cocaïne affecte la signalisation des H3R et mGlu1/5R de manière différente de son impacte sur leur signalisation croisée. Nos résultats démontrent l'existence d'une régulation croisée de certains RCPG dans le cerveau de rat. De plus, la consommation chronique de la cocaïne affecte différemment la signalisation induite par l'activation d'un récepteur et l'induction d'une signalisation croisée<br>Alterations of neuronal activity, mediated by G-protein coupled receptors (GPCRs), can modulate neuronal circuits and are thought to be important in the development and expression of diseases as cocaine addiction. GPCR activity is regulated by various mechanisms, including protein-protein interactions in the membrane, permitting these receptors to crosstalk and form homo-and heteromers. Cocaine blocks monoamine reuptake leading to increased synaptic presence of various neurotransmitters including dopamine, histamine and glutamate in the brain. First we describe that dopamine D1 receptors (D1R) and histamine H3 receptors (H3R) crosstalk in the rodent brain. Chronic cocaine self-administration altered the crosstalk between D1R and H3R in the striatum, a brain structure involved in habit learning and motor control. The altered signaling was observed in both individual receptor signaling and by D1R-H3R crosstalk signaling. Both histamine H¬3R and metabotropic glutamate 1/5 receptors (mGlu1/5R) are highly expressed in the hippocampus and the striatum of rodents and they are involved in behaviors regulated by these structures. We describe that H3R and mGlu1/5R crosstalk in pyramidal neurons of the hippocampus and in the striatum of rats. In addition, we found that signaling through H3R and mGlu1/5R were differently affected by chronic cocaine self-administration than the apparent crosstalk between the receptors. These results show evidence of GPCR interactions in adult rodent brain and reveal that chronic cocaine self-administration differently affected crosstalk and single receptor mediated signaling

<p>Drug delivery to the nasal and ocular mucosa faces several obstacles. One of these is from the effective clearance mechanisms present in the nose and eye. Polymer gels with suitable rheological properties can facilitate the absorption of poorly absorbed drugs by increasing the contact time of the drug with the mucosa. This has been attributed to the rheological and mucoadhesive properties of the gel. The main objective of this thesis was to investigate the importance of these features for the anticipated in vivo contact time, here exemplified by the ocular and nasal routes of administration.</p><p>The in situ gelling polymer gellan gum was found to have a favourable rheological and in vivo performance. When administered in the nasal cavity of rats, a gel was formed that could remain at the site of administration for up to 4 hours. In addition, the epithelial uptake and transfer of a 3 kDa fluorescein dextran was higher than for a mannitol solution. Therefore, it was concluded that a gellan gum formulation should be a promising strategy for nasal drug delivery.</p><p>The potential mucoadhesive properties of a variety of polymer gels were investigated using a rheological method and by measuring the tensile force required to detach the gel from a mucosa. With both methods the rheological properties of the gel were a determining factor for the results obtained. The rheological method was found to have several limitations. One of these was that a positive response, interpreted as mucoadhesion, was only seen with weak gels. The tensile method could, in contrast, detect strengthening of the mucus only for strong gels. However, this method reflects the in vivo performance of the gel better than the rheological method.</p><p>Finally, dielectric spectroscopy was explored as a tool for investigating the likelihood of intimate surface contact between the gel and the mucus layer. This novel approach involved determining the ease with which a charged particle can pass the gel-mucus interface layer, and may enable the study of the events at the interface closer to the molecular level, than is possible with the rheological and tensile strength methods.</p>

The putative 5-HT6 receptor agonist ST1936 has been shown to increase extracellular dopamine (DA) in the n.accumbens (NAc) Shell and in the medial prefrontal cortex (PFCX). These observations suggest that 5-HT6 receptors modulate DA transmission in mesolimbic and mesocortical terminal DA areas. To investigate the behavioral counterpart of this interaction I studied in rats the effect of 5-HT6 receptor blockade on cocaine stimulated overflow of DA in dialysates from the PFCX and from the NAc Shell and on cocaine i.v. selfadministration. Pretreatment with the 5-HT6 antagonist SB271046 reduced cocaine-induced increase of dialysate DA in the NAc Shell but not in the PFCX and impaired i.v. cocaine selfadministration. These suggest that 5-HT6 receptors play a role in cocaine reinforcement via their facilitatore interaction with DA projections to the NAc Shell. This 5-HT/DA interaction might provide the basis for a new pharmacotherapeutic strategy of cocaine addiction. Caffeine is one of the psychoactive substances most widely used as adulterant in illicit drugs, such as cocaine. Animal studies have demonstrated that caffeine is able to potentiate cocaine actions, although the enhancement of the cocaine reinforcing property by caffeine is less reported, and the results depend on the paradigms and experimental protocols used. In the present study I examined the ability of caffeine to enhance the motivational and rewarding properties of cocaine using the intravenous self-administration paradigm in rats. Additionally, the role of caffeine as a primer cue during extinction was evaluated. To this end, we assessed in naïve rats: 1) the ability of the combination of cocaine (0,125 mg/kg/infusion) and caffeine (0,0625 mg/kg/infusion) to maintain self-administration in fixed ratio (FR) and progressive ratio (PR) schedules of reinforcement compared with cocaine and caffeine alone; 2) the effect of caffeine in the maintenance of responding in the animals exposed to the combination of the drugs during cocaine extinction. Cocaine and the combination of cocaine and caffeine were self-administered on a FR and PR schedules of reinforcement, and the responding for the combination of the drugs was higher than cocaine alone. Caffeine was not reliably self-administered, but was able to maintain a drug-seeking behavior in rats previously exposed to cocaine plus caffeine. These findings suggest that the presence of caffeine enhances the reinforcing effects of cocaine and the motivational value of the drug. Our results highlight the role of active adulterants commonly used in illicit street drugs.