Licentiatavhandling (sammanfattning) Linköping : Linköpings universitet, 2009. / Härtill 2 uppsatser.

Thesis (Ph.D.) - University of St Andrews, April 2008.

Thesis (M.SC.) - Carleton University, 2007. / Includes bibliographical references (p. 59-94). Also available in electronic format on the Internet.

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Agradecemos a compreensão. on 2016-08-03T13:36:52Z (GMT) / Submitted by PAULA LAZILHA FALEIROS null (paulal.faleiros@hotmail.com) on 2016-08-03T13:55:04Z No. of bitstreams: 1 TESE DE DOUTORADO - PAULA FINAL.pdf: 7377120 bytes, checksum: c55964dddab287b8b096d337e07195bf (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-08-03T18:08:55Z (GMT) No. of bitstreams: 1 faleiros_pl_dr_araca_par.pdf: 1921873 bytes, checksum: 138048a39be83d82924179cc13951f8c (MD5) / Made available in DSpace on 2016-08-03T18:08:55Z (GMT). No. of bitstreams: 1 faleiros_pl_dr_araca_par.pdf: 1921873 bytes, checksum: 138048a39be83d82924179cc13951f8c (MD5) Previous issue date: 2016-07-05 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Objetivo: Avaliar do ponto de vista biomecânico, histomorfométrico, histoquímico e através de marcadores imunoistoquímicos a influência da terapia com laser em baixa intensidade (LLLT) na osseointegração de implantes instalados em tíbias de ratos modificados ou não sistemicamente pela nicotina. Materiais e métodos: Um total de 120 ratos (Wistar) foi distribuído em dois grandes grupos experimentais (n = 60) submetidos a duas aplicações diárias de solução salina (Veh) ou nicotina (Nic). Após 30 dias, implantes de titânio (2.2 mm x 4 mm) foram instalados na metáfise proximal de ambas as tíbias de todos os animais. Grupos experimentais de diferentes tratamentos locais do alvéolo cirúrgico previamente a instalação do implante (n = 30) foram criados no momento da cirurgia: Veh - nenhum tratamento; Veh/LLLT - irradiação do alvéolo cirúrgico com laser em baixa intensidade; Nic - nenhum tratamento; Nic/LLLT - irradiação do alvéolo cirúrgico com laser em baixa intensidade. O laser utilizado foi o Thera Lase (InGaAlP 660 nm, modo de aplicação contínuo, em contato com a área, 35 mW, 0,14 J, 4,9 J/cm2, por 4 segundos). Dez animais de cada grupo foram eutanasiados aos 15, 30 e 60 dias pós-operatórios. As tíbias direitas foram submetidas à análise de torque reverso e posteriormente processadas para confecção de cortes histológicos descalcificados, corados por hematoxilina e eosina, vermelho picrosirius ou submetidos a reações imunoistoquímicas com os anticorpos primários policlonais: HIF-1 α, VEGF, BMP-2, RUNX-2, OCN, ALP, RANKL, OPG e TRAP. A partir das tíbias esquerdas foram preparados cortes histológicos calcificados para análise das porcentagens de contato osso-implante (BIC) e área óssea (BA), dentro dos limites das roscas do implante. A análise foi realizada através da microscopia de luz polarizada para análise histoquímica e da microscopia óptica de campo claro para a análise histológica, histométrica e imunoistoquímicas. As imunomarcações foram submetidas à análise qualitativa para os anticorpos HIF-1 α, VEGF, BMP-2, OCN, ALP, RANKL e OPG e análise quantitativa para os anticorpos RUNX-2 e TRAP. Os dados quantitativos foram analisados estatisticamente (p≤0,05). Resultados: A nicotina atrasou a produção de elementos da matriz óssea, diminuiu o padrão de imunomarcação de HIF-1α, VEGF, BMP-2, OCN, ALP e OPG, aumentou o padrão de imunomarcação de RANKL e aumentou a quantidade de células imunorreativas a RUNX-2 e TRAP, diminuindo a BA, embora não tenha sido capaz de influenciar o torque de remoção e o BIC. O laser aumentou o padrão de imunomarcação de HIF-1α, VEGF, BMP-2, OCN e ALP, aumentou a quantidade de células imunorreativas a RUNX-2 e TRAP, aumentou a BA, mas também não foi capaz de influenciar o torque de remoção e o BIC. Adicionalmente, o laser influenciou positivamente a BA e o torque de remoção dos implantes, aumentou a angiogênese e a diferenciação osteoblástica, promovendo a formação óssea, biomineralização e maturação óssea peri-implantar nos animais modificados sistemicamente pela nicotina. Conclusão: A LLLT é capaz de promover o processo de reparo ósseo peri-implantar em condições normais e compensar os efeitos negativos da nicotina na osseointegração. / Objective: To evaluate, from biomechanical, histomorphometric, histochemical stand point and through of immunohistochemical markers, the influence of low level laser therapy (LLLT) on the osseointegration of implants placed in the tibias of nicotine systemically modified (or not) rats. Materials and methods: A total of 120 rats (Wistar) was assigned into two major experimental groups (n = 60) underwent two-daily-applications of saline (Veh) or nicotine (Nic). After thirty days, titanium implants (2.2 mm x 4 mm) were placed in the proximal metaphysis of both tibiae from all animals. Experimental groups of different surgical alveolus local treatments prior to implant placement (n = 30) were created at the moment of the surgery: Veh - no local treatment; Veh/LLLT - irradiation of surgical alveoli with low level laser; Nic - no local treatment; Nic/LLLT - irradiation of surgical alveoli with low level laser. The laser used was Thera Lase® (InGaAIP 660 nm, in continuous mode, in contact with the area, 35 mW, 0.14 J, 4.9 J/cm2 , for 4 seconds). Ten animals from each group were euthanized at 15, 30 and 60 days postoperative. The right tibiae were submitted to reverse-torque analysis and, then processing in order to prepare decalcified histological sections, stained either by hematoxylin and eosin or picrosirius red or subjected to immunohistochemical reactions with the primary polyclonal antibodies: HIF-1 α, VEGF, BMP-2, RUNX-2, OCN, ALP, RANKL, OPG and TRAP. From left tibias were prepared undecalcified histological sections to evaluate the percentages of bone-to-implant contact (BIC) and bone area (BA) within the limits of the implant threads. The histochemical analysis was performed by a polarized light microscopy and the histological, histometric and immunohistochemical analysis were performed by a bright field optical microscopy. The HIF- 1α, VEGF, BMP-2, OCN, ALP, RANKL and OPG immunostaining were submitted to qualitative analysis and the RUNX-2 and TRAP immunostaining were submitted to quantitative analysis. Quantitative data were analyzed statistically (p≤0.05). Results: The nicotine delayed the production of bone matrix components, decreased the imunolabeling pattern of HIF-1α, VEGF, BMP-2, OCN, ALP and OPG, increased the imunolabeling pattern of RANKL, increased the amount of RUNX-2 and TRAP-immunoreactive cells, reduced the BA, but did not influence the BIC and the force required to break the osseointegration. The laser increased the imunolabeling pattern of HIF-1α, VEGF, BMP-2, OCN and ALP, increased the amount of RUNX-2 and TRAP-immunoreactive cells, increased the BA, but also was not be able to influence the BIC and the force required to break the osseointegration. In addition, the laser positively influenced the BA and the implant removal torque, increased the angiogenesis and osteoblast differentiation, promoted bone formation, biomineralization and peri-implant bone maturation in the nicotine systemically modified rats. Conclusions: LLLT is able to promote the peri-implant bone repair process in normal conditions and is able to compensate the negative effects of nicotine on osseointegration. / FAPESP: 2013/11863-0

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Uma questão particularmente relevante é o fato de exposições precoces a drogas de abuso durante o desenvolvimento potencialmente aumentarem a susceptibilidade a estas drogas posteriormente durante o desenvolvimento. No presente estudo utilizando estudos comportamentais e eletrofisiológicos, investigamos efeitos tardios da exposição de camundongos à fumaça de cigarro, à nicotina e ao etanol durante o período que corresponde à gestação em humanos. Para tal, esta tese foi dividida em 2 estudos. No Estudo 1, submetemos camundongos durante o período que corresponde à gestação de humanos à fumaça de cigarro e/ou etanol visando investigar se a estas drogas de abuso, separadamente ou quando combinadas, programam maior susceptibilidade aos efeitos da nicotina durante a adolescência (PN30) ou idade adulta (PN90). Para avaliar a susceptibilidade, utilizamos 3 testes: campo aberto (CA), preferencia pela nicotina (PPN) e preferencia condicionada por lugar (CPP). No Estudo 2, os animais foram expostos a nicotina durante o período gestacional e, no período que corresponde à infância (PN9 a PN20), fatias de cérebro contendo o núcleo tegumental laterodorsal (LDT) foram expostas a etanol. Este núcleo foi escolhido uma vez que estudos recentes indicam sua participação em mecanismos de toxicodependência. Foram realizados registros eletrofisiológicos de uma única célula. No Estudo 1, identificamos maior sensibilidade para os efeitos da reexposição à nicotina na adolescência quando comparada com a idade adulta . Em animais testados no CA durante a adolescência, a nicotina foi capaz de causar aumento da atividade locomotora nos animais controle, previamente expostos à fumaça de cigarro e ao etanol. Contudo, em animais expostos à fumaça combinada com etanol, não houve aumento da locomoção. Na idade adulta, a nicotina causou um aumento da atividade locomotora no CA somente nos animais expostos à fumaça de cigarro. Quanto ao CPP, a exposição prévia à fumaça de cigarro e ao etanol causaram aumento da resposta condicionada à nicotina em fêmeas adolescentes. Nos animais previamente expostos à combinação entre fumaça de cigarro e etanol, a resposta condicionada à nicotina não atingiu significância estatística. Não houve alterações na idade adulta. A exposição a fumaça de cigarro e/ou etanol não afetou a PPN. No Estudo 2, os dados eletrofisiológicos mostraram que a exposição pré-natal à nicotina foi capaz de alterar as correntes de despolarização basais e o potencial de repouso de células do LDT A nicotina também foi capaz de alterar as respostas deste núcleo ao etanol reduzindo as correntes de despolarização e aumentando, embora que não de forma significativa, as correntes inibitórias. De acordo com estes dados, injurias causadas pela exposição à fumaça do cigarro, à nicotina isoladamente, e ao etanol durante o desenvolvimento são capazes de perdurar por um logo tempo na vida do individuo, alterando as respostas a comportamentais e celulares a uma exposição tardia à nicotina e ao etanol. / Particularly relevant is the fact that early exposure to drugs of abuse during development potentially increases drug susceptibility later during development. In the present study we used mice models to investigate postnatal behavioral and electrophysiological effects of exposure to cigarette smoke, nicotine and ethanol during the period that corresponds to pregnancy in humans. To this end, this thesis was divided into two studies. In Study 1, we submitted mice to cigarette smoke and / or ethanol in order during the period that corresponds to human pregnancy to investigate whether these drugs of abuse, alone or when combined, programs increased susceptibility to the effects of nicotine during adolescence (PN30) or adulthood (PN90). To evaluate susceptibility, we use three tests: open field (OF), preference for nicotine (PFN) and conditioned place preference (CPP). In Study 2, the animals were exposed to nicotine during pregnancy and, in the period corresponding to childhood (PN9 to PN20), brain slices containing the laterodorsal tegmental nucleus (LDT) were exposed to ethanol. This nucleus was chosen based on recent studies that indicate that it participates in mechanisms of addiction. Whole cell patch clamp recordings were performed. In Study 1, a higher sensitivity to the effects of nicotine exposure was identified during adolescence when compared to adulthood. In animals tested in the OF during adolescence, nicotine was able to cause an increase in locomotor activity in controls, in mice previously exposed to cigarette smoke, and in those exposed to ethanol. However, nicotine failed to increase locomotion in mice previously exposed to smoke combined with ethanol. In adulthood, nicotine caused an increase in OF locomotor activity only in animals exposed to cigarette smoke. In the CPP, previous exposure to cigarette smoke, and to ethanol caused an increase of the conditioned response to nicotine in adolescent females. In animals previously exposed to the combined cigarette smoke and ethanol, the conditioned response to nicotine did not reach statistical significance. There were no changes in adult mice. Exposure to cigarette smoke and / or ethanol did not affect the PFN. In Study 2, electrophysiological data have shown that prenatal exposure to nicotine alters the pattern of basal and depolarization of the resting potential of cells LDT. Nicotine was also able to change the answers this core ethanol reducing and increasing depolarization currents, although not significantly inhibitory currents. According to these data, injuries caused by exposure to cigarette smoke, nicotine alone, and ethanol during development persist during postnatal development, changing the behavioral and cellular responses to a late exposure to nicotine and ethanol.

abstract: Smoking remains the leading cause of preventable death in the United States, and early initiation is associated with greater difficulty quitting. Among adolescent smokers, those with attention-deficit hyperactivity disorder (ADHD), characterized by difficulties associated with impulsivity, hyperactivity, and inattention, smoke at nearly twice the rate of their peers. Although cigarette smoking is highly addictive, nicotine is a relatively weak primary reinforcer, spurring research on other potential targets that may maintain smoking, including the potential benefits of nicotine on attention, inhibition, and reinforcer efficacy. The present study employs the most prevalent rodent model of ADHD, the spontaneously hypertensive rat (SHR) and its control comparison Wistar Kyoto (WKY) to examine the effects of acute and chronic subcutaneous nicotine injections on performance in three operant response inhibition paradigms. Functional activation in select regions of the prefrontal cortex and striatum was also explored. Acute (0.1, 0.3, 0.6 mg/kg) and chronic (0.3 mg/kg) nicotine increased impulsive responding regardless of strain, dose, or operant schedule. Dose-dependent decreases in latency to initiate the task were also observed. SHR receiving daily nicotine injections showed less activation in the nucleus accumbens shell compared to saline controls. Despite close similarities, one of the three operant tasks did not detect response inhibition deficits in SHR relative to WKY. A closer examination of these tasks may highlight critical components involved in the amelioration of response inhibition deficits. / Dissertation/Thesis / Doctoral Dissertation Psychology 2014

abstract: Tobacco and alcohol are the most commonly abused drugs worldwide. Many people smoke and drink together, but the mechanisms of this nicotine (NIC) -ethanol (EtOH) dependence are not fully known. EtOH has been shown to affect some nicotinic acetylcholine receptors (nAChRs), which potentially underlies NIC-EtOH codependence. Ventral Tegmental Area (VTA) dopamine (DA) and γ-aminobutyric acid (GABA) neurons express different nAChR subtypes, whose net activation results in enhancement of DA release in the Prefrontal Cortex (PFC) and Nucleus Accumbens (NAc). Enhancement of DA transmission in this mesocorticolimbic system is thought to lead to rewarding properties of EtOH and NIC, clarification of which is relevant to public health and clinical diseases. The aim of this study was to elucidate pharmacological mechanisms of action employed by both NIC and EtOH through nAChRs in VTA neurons by evaluating behavioral, network, synaptic and receptor functions therein. It was hypothesized that VTA GABA neurons are controlled by α7 nAChRs on presynaptic GLUergic terminals and α6 nAChRs on presynaptic GABAergic terminals. NIC and EtOH, via these nAChRs, modulate VTA GABA neuronal function. This modulation may underlie NIC and EtOH reward and reinforcement, while pharmacological manipulation of these nAChRs may be a therapeutic strategy to treat NIC or EtOH dependence. This data demonstrates that in VTA GABA neurons, α7 nAChRs on GLUergic terminals play a key role in the mediation of local NIC-induced firing increase. α6*-nAChRs on GABA terminals enhances presynaptic GABA release, and leads to greater inhibition to VTA GABA neurons, which results in an increase VTA DA neuron firing via a disinhibition mechanism. Genetic knockout of these nAChRs significantly prevents EtOH-induced animal conditioned place preference (CPP). Furthermore, levo-tetrahydropalmadine (l-THP), a compound purified from natural Chinese herbs, blocks nAChRs, prevents NIC-induced DA neuronal firing, and eliminates NIC CPP, suggesting it as a promising candidate in a new generation of interventions for smoking cessation. Improved understanding of underlying mechanisms and development of new drugs will increase the number of successful quitters each year and dramatically improve the quality of life for millions suffering from addiction, as well as those around them. / Dissertation/Thesis / Doctoral Dissertation Neuroscience 2015

abstract: The ability to manipulate the interaction between small molecules and biological macromolecules towards the study of disease pathogenesis has become a very important part of research towards treatment options for various diseases. The work described here shows both the use of DNA oligonucleotides as carriers for a nicotine hapten small molecule, and the use of microsomes to study the stability of compounds derived to treat mitochondrial diseases. Nicotine addiction is a worldwide epidemic because nicotine is one of the most widely used addictive substances. It is linked to early death, typically in the form of heart or lung disease. A new vaccine conjugate against nicotine held within a DNA tetrahedron delivery system has been studied. For this purpose, several strands of DNA, conjugated with a modified dTpT having three or six carbon atom alkynyl linkers, have been synthesized. These strands have later been conjugated to three separate hapten small molecules to analyze which conjugates formed would be optimal for further testing in vivo. Mitochondrial diseases are hard to treat, given that there are so many different variations to treat. There is no one compound that can treat all mitochondrial and neurodegenerative diseases; however, improvements can be made to compounds currently under study to improve the conditions of those afflicted. A significant issue leading to compounds failing in clinical trials is insufficient metabolic stability. Many compounds have good biological activity, but once introduced to an animal, are not stable enough to have any effect. Here, several synthesized compounds have been evaluated for metabolic stability, and several showed improved stability, while maintaining biological activity. / Dissertation/Thesis / Doctoral Dissertation Chemistry 2016

Este trabalho visa avançar o conhecimento sobre a interferência da nicotina no protocolo de treino forçado em gerbilos isquêmicos. Utilizamos 110 Gerbilos, distribuídos em 11 grupos. Grupo controle com animais ingênuos (C), falso operado (S) e isquêmicos (I). Grupos nicotina (CN, SN, SNE, IN e INE) e salina (CS, SS, IS) com ou sem treinamento. Os animais receberam uma injeção de 2mg/kg de nicotina ou salina durante 9 dias. Utilizamos uma esteira motorizada (treino contínuo, 5 dias, 5m/min por 15 minutos), um monitor de atividades e o Rota Rod. Os resultados foram analisados por uma ANOVA e pós teste de Holm-Sidak, p<0,05. Observamos aumento de apresentação de todos comportamentos no monitor de atividades, o CN maior do que os C, S, SS, SNE e INE, o SN maior do que o SNE, o I maior do que os C, S, SS, SNE, IS, INE e ainda, o IN maior do que os C, S, SS, SNE, IS e INE (cruzamento, F10,93 = 2,97), (distância, F10,93 = 2,79), (velocidade, F10,93 = 2,79) e (atividade, F10,93 = 2,81). No Rota Rod o CN apresentou maior tempo de permanência com relação aos outros grupos exceto SNE, já o SNE, em relação aos CS, S, SN, I, IN e o I menor tempo de permanência do que o C e INE (F10,93 = 3,35). Nos grupos I e IS observamos menor densidade de neurônios no hipocampo (F6,42 = 31,02), córtex motor M1 (F6,42 = 4,01) e estriado (F6,42 = 23,33). A nicotina não interferiu com a melhora do comportamento dos animais isquêmicos. / This work aims to advance the knowledge about the interference of nicotine in the forced training protocol in ischemic gerbils. We use 110 gerbils, distributed in 11 groups. Control (C), false operated (S) and ischemic (I). Nicotine groups (CN, SN, SNE, IN INE) and sham (CS, SS, SI) with or without training. The animals received an injection of 2mg/kg of nicotine or saline for 9 days. We use a treadmill (continuous training 5 days, 5m / min for 15 minutes), an activity monitor and the Rota Rod (RR). The results were analyzed by an ANOVA and post Holm-Sidak test, p <0.05. We observed increased in all behaviors in the activity monitor, increase of CN than C, S, SS and NSS INE, SN increase compared to NES, the I increase than C, S, SS, ENS, S, NSI and also the IN showed an increase compared to C, S, SS, SNE, SI and NSI (crossing, F10,93 = 2,97), (distance, F10,93 = 2,79), (speed, F10,93 = 2,79) and (activity, F10,93 = 2,81). In RR, CN showed an increase of time spent with the other groups except SNE, and an increase in the SNE, compared to the CS, S, SN, I, IN and I showed an decrease of time compared to C and INE (F10,93 = 3,35). In groups I and and IS we observed an decrease of neurons density in the hippocampus (F6,42 = 31,02), motor cortex M1 (F6,42 = 4,01) and striatum (F6,42 = 23,33). Nicotine did not interfere with the improvement of motor behavior in ischemic animals.

Made available in DSpace on 2014-06-11T19:22:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-12-11Bitstream added on 2014-06-13T20:09:54Z : No. of bitstreams: 1 manzano_bm_me_prud.pdf: 326376 bytes, checksum: 0b7d0d1cb3bdf834e64a73541c8ddedd (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O tabagismo é considerado um dos principais fatores de risco modificáveis de doenças cardiovasculares e suas complicações, dentre as quais doença vascular aterosclerótica, hipertensão, infarto do miocárdio, angina instável e morte súbita. Os efeitos cardiovasculares promovidos pela nicotina ocorrem principalmente, devido ao aumento da atividade simpática, decorrente do estímulo de liberação de catecolaminas, por meio da ativação dos receptores nicotínicos localizados nas terminações nervosas simpáticas pós-ganglionares periféricas e medula adrenal. Além disso, o fumo acarretada uma disfunção autonômica a qual pode ser avaliada por meio da variabilidade da freqüência cardíaca (VFC) que descreve as oscilações dos intervalos entre batimentos consecutivos (intervalos RR) e reflete a atividade do sistema nervoso autônomo (SNA) sobre o nódulo sinusal, sendo uma ferramenta clínica para avaliar e identificar comprometimentos na saúde. Estudos demonstram que o tabagismo crônico leva a ativação simpática e redução da modulação vagal, de forma que essas alterações autonômicas basicamente se expressam por diminuição dos índices de VFC em fumantes. A redução da VFC é considerada uma condição de alta morbidade e mortalidade cardíaca, no entanto, alguns estudos apontam que a cessação do tabagismo pode levar a restauração da função autonômica... / Smoking is considered an important modifiable risk factor for cardiovascular disease and its complications, such as atherosclerosis, hypertension, myocardial infarction, instable angina and sudden death. The cardiovascular effects promoted by nicotine are caused by increased sympathetic activity occurred because of catecholamine release by nicotinic receptors activation at peripheral postganglionic sympathetic nerve endings and adrenal medulla. Moreover, smoking leads to autonomic dysfunction that can be evaluated by heart rate variability (HRV) which describes the oscillations in the interval between consecutive heart beats (RR interval) and reflects the autonomic nervous system (ANS) activity on the sinus node and as a clinical instrument to assess and identify health involvements. Studies have been demonstrated that chronic smoking causes sympathetic activation and reduces vagal modulation and that autonomic alterations basically are showed by decreases of HRV indices in smokers... (Complete abstract click electronic access below)