Local infiltration analgesia (LIA) is a new technique for postoperative pain management following knee arthroplasty. LIA involves a long-acting local anesthetic (ropivacaine), a non-steroid anti-inflammatory drug (ketorolac) and epinephrine infiltrated into the knee joint during surgery and injected postoperatively via a catheter. In the first two studies, LIA was compared with placebo in unicompartmental (I) and total (II) knee arthroplasty. Postoperative pain levels, morphine consumption and the incidence of side effects were lower in the LIA groups. In addition, we found a shorter length of hospital stay in the LIA group following unicompartmental knee arthroplasty compared with placebo (I), while the time to home readiness was shorter in the LIA group following total knee arthroplasty (II). In this study, we found that the unbound venous blood concentration of ropivacaine was below systemic toxic blood concentrations in a sub-group of patients. In the third study, LIA was compared with intrathecal morphine for postoperative pain relief following total knee arthroplasty (III). Pain scores and morphine consumption were lower, length of hospital stay was shorter and patient satisfaction was higher in the LIA group. In the final study, we investigated the effect of minimally invasive surgery (MIS) compared with conventional surgery in unicompartmental knee arthroplasty (IV). Both groups received LIA. We found no statistically significant differences in postoperative pain, morphine consumption, knee function, home readiness, hospital stay or patient satisfaction. In conclusion, LIA provided better postoperative pain relief and earlier mobilization than placebo, both in unicompartmental and total knee arthroplasty. When compared to intrathecal morphine, LIA also resulted in improved postoperative pain relief and earlier mobilization. Minimally invasive surgery did not improve outcomes after unicompartmental knee arthroplasty, when both groups received LIA.

Posttraumatic Stress Disorder (PTSD) is an anxiety disorder that affects over 7.7 million adults and carries an estimated societal cost of $3.1 billion every year. People develop PTSD after exposure to a traumatic event. Alone or combined, approved pharmacotherapies or psychotherapy are somewhat effective, but symptoms for many remain refractory. Emerging evidence suggests that opiate systems may modulate the development and expression of PTSD, and their role can be investigated preclinically. Pavlovian fear conditioning is a preclinical model which elicits behaviors mirroring those that occur in humans during and after exposure to trauma. This presents an experimental tool that can help elucidate the opiate mechanisms involved in traumatic memory as well as the resulting fear behavior. Mu opioid receptor (MOR) analgesics, such as morphine, are often given as a response to trauma, and there is emerging evidence that they are, at least partially, protective against PTSD. The kappa opioid receptor (KOR) system has also been implicated in stress-related processes, with KOR agonists reported to enhance stress in both laboratory animals and in humans, and KOR antagonists reported to attenuate stress-like behaviors preclinically. This project attempted to clarify part of the role of the mu and kappa opiate receptor systems in mediating effects of Pavlovian fear conditioning in mice as a predictor of their involvement in some of the signs and symptoms of PTSD. Kappa agonists increased acute fear responses but surprisingly also facilitated fear extinction learning. This would suggest that the use of kappa agonists might increase the efficiency and effectiveness of this therapy and could improve existing PTSD patient outcomes. MOR agonists, as well as KOR antagonists reduced acute and long-term fear behavior. These results support that the use KOR analgesics like morphine and fentanyl in the treatment of trauma could have an added benefit of reducing the emergence and persistence of PTSD. Self-medication may help explain the comorbidity of opioid abuse in PTSD patient populations. Understanding the relative effects of these opiate ligands could lead to more informed usage of MOR analgesics which vary in mu and kappa receptor activity under battlefield and other traumatic conditions.

INTRODUÇÃO: Baseados em estudo que evidenciou menor área de infarto do miocárdio (IM) em ratos submetidos a treinamento físico (TF),na ausência de reperfusão; e na liberação de endorfinas que ocorre durante o TF, nossos objetivos são: demonstrar se não só TF, mas também opióides e isquemia/reperfusão (IR) intermitente são capazes de reduzir área de IM, na ausência de reperfusão; se TF e opióides exibem efeito sinérgico e se o mecanismo de redução da área de IM pelo TF envolve receptores opióides. MATERIAIS E MÉTODOS: Ratos Wistar machos (n=76) foram divididos em 7 grupos:1- controle;2- TF (esteira elétrica,1 hora/dia,5 vezes/semana,por 12 semanas), antes do IM; 3- morfina antes do IM; 4- morfina+TF; 5- grupo com 3 ciclos de IR antes do IM; 6- naloxone antes da morfina; 7- naloxone antes de cada dia de TF. Todos os ratos foram submetidos à mensuração da pressão diastólica final (PDF) e a IM através da oclusão da artéria descendente anterior. A eficácia do TF foi avaliada através do consumo de oxigênio (VO2) e da distância máxima percorrida. Os ratos foram sacrificados no 8o pós-IM e a área de IM mensurada por planimetria. RESULTADOS: Não houve diferença no peso inicial (p=0,94), mortalidade (p=0,99), e relação peso cardíaco/peso corporal (p=0,29) entre os grupos. Entretanto, houve aumento do deltaVO2 (VO2 pico - VO2repouso) (p=0,0001)e da distância máxima percorrida (p=0,0001), nos grupos treinados. A PDF aumentou no pós-IM, em todos os grupos (p=0,0001). Os grupos tratados tiveram menor área de IM (p=0,0001), com exceção dos grupos morfina + naloxone e TF+ naloxone sendo que não houve maior redução no grupo TF+morfina. Os grupos TF e TF+morfina apresentaram maior espessura do septo inter-ventricular, em relação ao grupo controle (p=0,0008). Já o grupo TF + naloxone não apresentou maior espessura do septo IV, em relação aos outros grupos. Também não houve diferença na densidade capilar (p=0,88). CONCLUSÃO: Não só TF, mas também morfina e IR reduzem a área de IM, na ausência de reperfusão, sendo que não há efeito sinérgico entre TF e morfina. Esta redução não ocorre através do aumento da densidade capilar. Além disto, a ação do TF sobre a área de IM provavelmente ocorre através do estímulo de receptores opióides, visto que seu bloqueio anulou o efeito cardioprotetor do TF / BACKGROUND AND OBJECTIVES: Studies have shown a decrease in infarcted area in rats submitted to exercise training (ET), in the absence of reperfusion. Based on that, we tested four hypotheses: 1- not only ET but also another stimulus that causes myocardial protection, like opioid infusion and brief periods of ischemia-reperfusion (IR) before irreversible left anterior descending (LAD) coronary occlusion could reduce infarct area, 2- ET plus opioid infusion could have additive effects in reducing infarct size, 3- blocking the opioid system we could lose the myocardial protection caused by ET, 4-myocardial protection given by different strategies could occur due to the increase in capillary density. METHODS: Male Wistar rats (n=76) were randomly assigned to 7 groups: control (n=11); ET (n=12); morphine infusion before myocardial infarction (MI) (n=14); ET plus morphine (n=11); naloxone (a non selective opioid receptor blocker) plus morphin (n=9); intermittent IR (n=12) before MI; naloxone before each ET session (n=7). All groups were submitted to MI by LAD ligation technique and to measurement of left ventricular end-diastolic pressure (LVEDP) before and 5 min after MI. ET was performed on a treadmill for 60 min, 5 times/week for 12 weeks at 60% peak oxygen (peak VO2). To evaluate the efficacy of ET, we tested the exercise capacity and the peak VO2 before and after experimental period. Seven days after MI induction, rats were killed and hearts were harvested. Infarct size was expressed by evaluation of necrotic area, expressed as a % of the risk region (total left ventricle area). RESULTS: There were no differences in initial weight, cardiac/animal weight or mortality among groups. Exercise training increased exercise capacity (p=0.0001) and delta VO2 (VO2 peak-VO2 rest) (p=0.0001). Inter-ventricular septum thickness was higher in the ET and ET plus morphine groups, compared to the control group (p=0.0008). The LVEDP increased in the post-MI period, for all groups (p=0.0001). All treatment groups but not morphine plus naloxone and ET plus naloxone showed a decrease in infarcted area (p=0.0001). There was no additional decrease in infarct size in the ET+ morphine group, comparing with each group alone . There was no difference in capillary density for all groups. CONCLUSION: Not only ET, but also morphine and IR decrease infarcted area, in the absence of reperfusion. There is no additional effect between ET and morphine. Moreover, this reduction is not due to an increase in capillary density. The effect of ET in decreasing infarct size might occur by opioid receptor stimulus

O tratamento com morfina ao final da prenhez, faz com que uma única dose dessa droga durante a lactação iniba o comportamento maternal e estimule a caça predatória. A intensidade dessa mudança comportamental depende da dose desafio de morfina utilizada. A exposição a drogas de abuso pode levar a um fenômeno denominado tolerância reversa, que consiste na exacerbação dos efeitos do tratamento agudo com um fármaco observada após a interrupção de um tratamento crônico. O objetivo deste estudo foi avaliar como os efeitos do pré-tratamento com morfina durante a gestação podem influenciar na seleção comportamental após o parto. Ratas foram expostas simultaneamente a filhotes e insetos, sendo observada a expressão dos comportamentos maternal e predatório. As ratas foram tratadas com injeções diárias de morfina (3,5 mg/Kg, s.c.) do 17º ao 21º dia de gestação, e desafiadas agudamente no 5o ou 6o dia de lactação com doses distintas de morfina (0,5, 1,0 e 1,5mg/Kg; grupos MM0,5, MM1,0 e MM1,5) ou salina (grupo MS). Grupos controle foram pré-tratados com salina e desafiados com morfina (0,5, 1,0 e 1,5mg/Kg; grupos SM0,5, SM1,0 e SM1,5) ou salina (grupo SS), respectivamente. Em seguida foram testadas no paradigma de escolha entre cuidar dos filhotes e caçar. Animais pré-tratados com morfina e desafiados com 1,0 mg/Kg tiveram parâmetros de comportamento maternal prejudicados e facilitação ao comportamento de caça de maneira significante, o que não foi observado nos animais desafiados com a dose de 0,5 mg/Kg. Animais desafiados com 1,5 mg/Kg de morfina tiveram prejuízo em relação ao comportamento maternal e facilitação do comportamento predatório tanto no grupo de fêmeas pré-tratadas com morfina, quanto com salina. Em ratas lactantes a exposição simultânea a filhotes e insetos permitiu revelar a existência de tolerância reversa à mudança comportamental induzida por estímulo opioidérgico. / Treatment of postpartum female rats with morphine inhibits maternal behavior. The same treatment also stimulates foraging in adult animals. Exposure to drugs of abuse may result in a progressive and enduring enhancement of their reinforcing effects. Puerperal treatment with morphine leads to reverse tolerance to this drug, ultimately influencing the effects of opiates on maternal behavior. The aim of the present study was to investigate whether abrupt withdrawal from repeated morphine treatment during late pregnancy may influence the effects of morphine on behavioral selection in lactating rats. Animals were exposed simultaneously to pups and insects, and the choice between taking care of the pups and hunting for insects was observed. Female Wistar rats were treated with morphine (3.5 mg/kg/day, subcutaneous [s.c.]) or saline for 5 days beginning on pregnancy day 17. On day 5 of lactation, animals were acutely challenged with morphine (0.5, 1.0, or 1.5 mg/kg, s.c.; MM0.5, MM1.0, and MM1.5 groups, respectively) or saline (MS group) and simultaneously tested for predatory hunting and maternal behavior. Control groups were pretreated with saline and challenged with morphine (SM0.5, SM1.0, and SM1.5 groups) or saline (SS group). Animals treated with morphine during late pregnancy and acutely challenged with 1.0 mg/kg morphine (MM1.0 group) exhibited significantly decreased maternal behavior and enhanced hunting. This effect was not evident for the 0.5 mg/kg dose. The 1.5 mg/kg morphine dose decreased maternal behavior and increased hunting in both the MM1.5 group and in animals acutely challenged with morphine after previous saline treatment (SM1.5 group). These results provide evidence of plasticity of the opioidergic role in behavioral selection during lactation.

Durante muito tempo a dor nos animais foi subestimada, pensando-se que não sentiam dor ou que seu limiar era alto. As manifestações de dor eram mal interpretadas ou ignoradas, e por este motivo não existia uma justificativa para a prevenção ou tratamento adequado da dor. Existem sinais de dor nos animais que podem passar desapercebidos, como sons e mudanças comportamentais ou gestuais que necessitam ser tomado em conta. Devemos observar tanto as variações de conduta como as fisiológicas, que nos indicam dor, para conseguir tratá-la adequada e oportunamente. Neste estudo foram utilizados 20 eqüinos encaminhados para cirurgia artroscópica junto ao Serviço de Cirurgia de Grandes Animais da FMVZ/USP, estabelecendo-se protocolos de avaliação de dor dos pacientes associados a tratamento da dor pós-operatória. Os animais foram divididos aleatoriamente em dois grupos, sendo que no grupo 1 foi aplicada morfina na dosagem 0,1 mg/kg, por via intra-articular ao término do procedimento artroscópico, e no grupo 2 administrou-se fenilbutazona na dose de 4.4 mg/kg, uma vez ao dia, durante três dias, sendo a primeira aplicação realizada imediatamente antes ao procedimento cirúrgico. Dez eqüinos, pertencentes aos Departamentos de Clínica Médica e de Cirurgia da FMVZ / USP, formaram o grupo controle para valores séricos de cortisol, que também foi mensurado nos grupos 1 e 2. Nestes animais não foi realizado procedimento artroscópico. Os animais operados que apresentaram dor acentuada, não controlada com o protocolo do experimento, receberam terapia analgésica adicional e foram então retirados da avaliação nos momentos seguintes. Os parâmetros observados para avaliação da dor foram: grau de claudicação, resposta à palpação local, escala numérica visual (ENV), escala facial de dor e mensuração de freqüência cardíaca, freqüência respiratória e cortisol sérico. Onze momentos foram avaliados durante cinco dias pela manhã e à tarde, e as duas, quatro e seis horas após o procedimento cirúrgico. A análise dos resultados baseou-se na comparação entre os grupos e entre os diferentes momentos. Os resultados analisados demonstraram que a avaliação comportamental e a mensuração da concentração de cortisol foram essenciais para complementar o exame habitualmente realizado através dos parâmetros fisiológicos, como freqüência cardíaca e respiratória. Para esta avaliação comportamental é necessário o uso de escalas que permitam fazer uma avaliação mais especifica para a espécie, portanto, a escala numérica visual e em especial a escala de dor pela face são uma boa maneira de avaliar a dor em cavalos e podem ser utilizadas rotineiramente para a determinação clínica de dor na espécie eqüina.Também pudemos observar que a utilização de fenilbutazona sistêmica foi mais efetiva no tratamento da dor após cirurgia artroscópica que a administração de morfina intra-articular, nas doses empregadas. / For a long time the pain in animals was underestimated, thought that they didnt feel pain or that its threshold was high. Demonstrations of pain were misinterpreted or ignored, for that reason there wasnt a justification for its prevention or adequate treatment. There are signs of pain in animals that can pass unnoticed, like sounds and behavioral or gesture changes that most be taken into account. We must to observe behavioral and physiological variations that would indicate existence of pain in animals in order to manage and treat it adequate and opportunely. In this study were used 20 horses referred for arthroscopic surgery to the Veterinary Hospital at FMVZ/USP, and were established protocols for the patient evaluation and treatment of postoperative pain. Horses were divided randomly in two groups: in the group 1, it was performed the intra-articular administration of morphine in a dose of 0,1mg/kg at the end of the arthroscopic procedure and in the group 2 It was administered phenilbutazone in a dose of 4.4mg/kg once a day for three days, being carried out the first application immediately before the surgical procedure. Ten horses, which belong to the Departments of Clinics and Surgery at FMVZ/USP, formed the control group for the seric cortisol levels determination that was also measured in groups 1 and 2. In these animals wasnt carried out any arthroscopic procedure. In specific cases when the horse presented intense pain that couldnt be controlled with the experimental protocol, it was instituted additional analgesia and the animal withdrawn of the subsequent evaluations. Parameters observed for pain evaluation were: degree of lameness, response to local palpation, numeric rating scale (NRS), scale of pain determined by the facial expression, measurement of respiratory and cardiac rate, and seric cortisol measurement. Eleven moments were evaluated during 5 days, in the morning and in the afternoon and at two, four and six hours after the surgical procedure. Results analysis was based on the comparison between the groups and among the different moments. Results of analysis showed that the behavioral evaluation and measurement of cortisol concentrations were essential to complement the routine exam carried out to obtain physiological parameters, as respiratory and cardiac rate. For the behavioral testing it is necessary the use of scales that allow to perform a more specific pain evaluation for the specie, for that purpose the visual analog scale and especially the scale of pain determined by the facial expression are a good way to evaluate the level of pain in horses, and they can be used in a routine way for the clinical pain assessment in horses. Also we could observe that the use of systemic phenilbutazone was more effective in the pain control after arthroscopic surgery that intra-articular administration of morphine at the end of the arthroscopic procedure in the doses used in the present study.

Dor em pediatria tem sido o foco de muitos estudos nas últimas décadas devido ao fato de que neonatos apresentam menor limiar a estímulos nocivos e inócuos em relação aos adultos. Desta forma, o uso de analgésicos é frequente para sedação e analgesia em UTIs pediátricas, e entre os opioides, a morfina é um dos mais utilizados. Adicionalmente, exposição a estímulos estressantes como a deprivação materna está entre os fatores ambientais relacionados a alterações no desenvolvimento neural. O estresse social ou a negligência do cuidado parental, e mais precisamente do cuidado materno em ratos, está associado a importantes alterações comportamentais na vida adulta. Entre estas alterações apresentadas ao longo da vida estão alterações na resposta ao estresse, e à alteração na sensibilidade a estímulos dolorosos, indexadas por hiperalgesia. Consquentemente, o estresse social gerado pela deprivação materna está relacionado a prejuízos cognitivos, emocionais e sociais, além de alterações neuroquímicas de longo prazo. Deste modo, é necessário atenção, prevenção e tratamento a esses eventos físicos, emocionais e comportamentais no período neonatal e durante a infância, uma vez que as bases neurobiológicas envolvidas nestes fenômenos ainda não foram completamente elucidadas. Considerando a relevância do tema, o objetivo deste estudo foi verificar os efeitos do tratamento com 5 μg de morfina, uma vez ao dia, do P8 ao P14 e a exposição a deprivação materna por 3 horas durante os primeiros 10 dias de vida em curto (P16), médio (P30) e longo prazo (P60), sobre o desenvolvimento dos reflexos neuromotores da prole por meio do Reflexo de Endireitamento, Geotaxia Negativa e Marcha; comportamento nociceptivo por meio dos testes Tail-Flick e Placa Quente, respectivamente. Um total de 58 filhotes foi utilizado. Os animais foram divididos em 5 grupos: controle total (C), que não recebeu nenhuma intervenção; salina (S), que recebeu solução salina; morfina (M), que recebeu morfina; deprivado salina (DS), que foram submetidos a deprivação maternal e receberam solução salina; e deprivado morfina (DM), que foram submetidos a deprivação maternal e receberam morfina. Em relação aos testes neuroquímicos, foram analisados níveis de BDNF, NGF, IL-1β e IL-4 em tronco e córtex cerebral que estão relacionados a fenômenos modulatórios em sistemas nervoso e imune. Os animais que receberam morfina e os deprivados maternos que receberam morfina apresentaram atraso no desenvolvimento dos reflexos iniciais. Alterações neuroquímicas também foram observadas. Os níveis de BDNF no tronco encefálico foram diminuídos em animais que receberam morfina e deprivação materna. Animais deprivados apresentaram um aumento nos níveis de NGF no tronco encefálico. Além disso, observou-se um aumento nos níveis de NGF do córtex cerebral em animais que receberam morfina, deprivados maternos e deprivados maternos que receberam morfina. Uma diminuição no limiar nociceptivo foi observada em animais que receberam morfina, deprivados maternos e os deprivados maternos que receberam morfina. Também houve interações em tronco encefálico e córtex cerebral nos níveis de BDNF, IL-1β e IL-4 entre as variáveis independentes: tratamento, deprivação e tempo, o que levou à modificação nos níveis centrais dos neuroimunomoduladores avaliados. Estes dados demonstram a importância de estudos focados nos efeitos do tratamento com morfina no período neonatal ao longo da vida, assim como na busca por alternativas terapêuticas que possam reverter possíveis alterações decorrentes da separação materna no período neonatal. / Pediatric pain has been the focus of many studies in the last decades due to the fact that neonates have a lower threshold for innocuous and noxious stimuli than for adults. Thus, the use of analgesics is frequent for sedation and analgesia in pediatric intensive care units, and among opioids, morphine is one of the most used. Additionally, exposure to stressful stimuli such as maternal deprivation is among the environmental factors related to changes in neural development. The social stress or neglect of parental care, and more precisely maternal care in rats, is associated with important behavioral changes in adult life. Among these changes presented throughout life are changes in the response to stress, and the change in sensitivity to painful stimuli, indexed by hyperalgesia. Consequently, the social stress generated by maternal deprivation is related to cognitive, emotional and social impairments, further to long-term neurochemical changes. Thus, attention, prevention and treatment are necessary to these physical, emotional and behavioral events in the neonatal period and during childhood, since the neurobiological bases involved in these phenomena have not yet been fully elucidated. Considering the relevance of the subject, the objective of this study was to verify the effects of treatment with 5 μg of morphine once a day from P8 to P14 and exposure to maternal deprivation for 3 hours during the first 10 days of short (P16), medium (P30) and long- term (P60), on the development of neuromotor reflexes of offspring through the righting Reflex, Negative Geotaxis and Gait; nociceptive behavior through the Tail-Flick and Hot Plate tests, respectively. A total of 58 puppies were utilized. The animals were divided in 5 groups: the total control group (C), which did not receive any intervention; saline group (S), which receive saline solution; morphine group (M), which receive morphine; deprived saline (DS), which were subjected to maternal deprivation and receive saline solution; and deprived morphine group (DM), which were subjected to maternal deprivation and receive morphine. In relation to the neurochemical tests, levels of BDNF, NGF, IL-1β and IL-4 were analyzed in the brainsteam and cerebral cortex and are related to modulatory phenomena of the nervous and immune systems. Animals that received morphine and deprived animais that received morphine showed a delay in the development of early reflexes. Neurochemical changes were also observed. BDNF levels in the brainstem were decreased in animals receiving morphine and maternal deprivation. Deprived animals had an increase in NGF levels in the brainstem. Besides, an increase in NGF levels of the cerebral cortex was observed in animals receiving morphine, maternal deprivation and maternal deprivation receiving morphine. A decrease in the nociceptive threshold was observed in animals receiving morphine, maternal deprivation, and maternal deprivation receiving morphine. There were also interactions in the brainstem and cerebral cortex in the levels of BDNF, IL-1β and IL-4 among the independent variables: treatment, deprivation and time, which led to the modification in the central levels of the neuroimmunomodulators evaluated. These data demonstrate the importance of studies focused on the effects of treatment with morphine in the neonatal period throughout life, as well as on the search for therapeutic alternatives that may reverse possible changes due to maternal deprivation in the neonatal period.

O tratamento com morfina ao final da prenhez, faz com que uma única dose dessa droga durante a lactação iniba o comportamento maternal e estimule a caça predatória. A intensidade dessa mudança comportamental depende da dose desafio de morfina utilizada. A exposição a drogas de abuso pode levar a um fenômeno denominado tolerância reversa, que consiste na exacerbação dos efeitos do tratamento agudo com um fármaco observada após a interrupção de um tratamento crônico. O objetivo deste estudo foi avaliar como os efeitos do pré-tratamento com morfina durante a gestação podem influenciar na seleção comportamental após o parto. Ratas foram expostas simultaneamente a filhotes e insetos, sendo observada a expressão dos comportamentos maternal e predatório. As ratas foram tratadas com injeções diárias de morfina (3,5 mg/Kg, s.c.) do 17º ao 21º dia de gestação, e desafiadas agudamente no 5o ou 6o dia de lactação com doses distintas de morfina (0,5, 1,0 e 1,5mg/Kg; grupos MM0,5, MM1,0 e MM1,5) ou salina (grupo MS). Grupos controle foram pré-tratados com salina e desafiados com morfina (0,5, 1,0 e 1,5mg/Kg; grupos SM0,5, SM1,0 e SM1,5) ou salina (grupo SS), respectivamente. Em seguida foram testadas no paradigma de escolha entre cuidar dos filhotes e caçar. Animais pré-tratados com morfina e desafiados com 1,0 mg/Kg tiveram parâmetros de comportamento maternal prejudicados e facilitação ao comportamento de caça de maneira significante, o que não foi observado nos animais desafiados com a dose de 0,5 mg/Kg. Animais desafiados com 1,5 mg/Kg de morfina tiveram prejuízo em relação ao comportamento maternal e facilitação do comportamento predatório tanto no grupo de fêmeas pré-tratadas com morfina, quanto com salina. Em ratas lactantes a exposição simultânea a filhotes e insetos permitiu revelar a existência de tolerância reversa à mudança comportamental induzida por estímulo opioidérgico. / Treatment of postpartum female rats with morphine inhibits maternal behavior. The same treatment also stimulates foraging in adult animals. Exposure to drugs of abuse may result in a progressive and enduring enhancement of their reinforcing effects. Puerperal treatment with morphine leads to reverse tolerance to this drug, ultimately influencing the effects of opiates on maternal behavior. The aim of the present study was to investigate whether abrupt withdrawal from repeated morphine treatment during late pregnancy may influence the effects of morphine on behavioral selection in lactating rats. Animals were exposed simultaneously to pups and insects, and the choice between taking care of the pups and hunting for insects was observed. Female Wistar rats were treated with morphine (3.5 mg/kg/day, subcutaneous [s.c.]) or saline for 5 days beginning on pregnancy day 17. On day 5 of lactation, animals were acutely challenged with morphine (0.5, 1.0, or 1.5 mg/kg, s.c.; MM0.5, MM1.0, and MM1.5 groups, respectively) or saline (MS group) and simultaneously tested for predatory hunting and maternal behavior. Control groups were pretreated with saline and challenged with morphine (SM0.5, SM1.0, and SM1.5 groups) or saline (SS group). Animals treated with morphine during late pregnancy and acutely challenged with 1.0 mg/kg morphine (MM1.0 group) exhibited significantly decreased maternal behavior and enhanced hunting. This effect was not evident for the 0.5 mg/kg dose. The 1.5 mg/kg morphine dose decreased maternal behavior and increased hunting in both the MM1.5 group and in animals acutely challenged with morphine after previous saline treatment (SM1.5 group). These results provide evidence of plasticity of the opioidergic role in behavioral selection during lactation.

Made available in DSpace on 2016-04-28T20:18:27Z (GMT). No. of bitstreams: 1 2008-Douglas dos Santos e Castro.pdf: 335295 bytes, checksum: b504eb6279e1e52327a6a9bf05f4f955 (MD5) Previous issue date: 2008-01-17 / The purpose of this study was to compare the effectiveness and duration of the analgesic tramadol to the morphine, administered by the epidural route in cats (Felis catus domesticus) For this, six females cats were used, without defined breed, weighing between three and four kilograms and age ranging from one to three years. After fasting food, in 12 hours and water, four hours, the animals were subjected to general anesthesia with isoflurane through an anesthetic box, in order to allow the completion of the epidural technique. Each animal received at random, in a blind study, three epidural route treatments at intervals of one week between them, with: saline solution to 0.9% in a volume equivalent to 0.22ml.kg-1 (Control Group), tramadol 1 mg.kg-1 diluted in saline solution and volume equivalent to 0.22 ml.kg-1 (Tramadol Group) and morphine 0.1mg.kg-1 diluted in saline solution and volume equivalent to 0.22ml.kg-1 (Morphine Group). One hour after administration of each drug and general anesthesia recovery, the animals were subjected to painful stimuli in three pre-defined places: in the dorsal tail basis, the side of the right and left thigh, giving continuity, 2, 3, 4, 6, 8, 10 and 12 hours respectively. The pain was qualified by the implementation of two Simple Descriptive Scales (SDS), a Visual Analog Scale (VAS), measurement of heart and respiratory rate. For qualitative variables used in the Latin Square design 3 x 3 with the Kruskal-Wallis test and for the quantitative variables used in Analysis of Variance (ANOVA). There was no difference in relation to the time of anesthesia and respiratory frequency, However, the heart rate was significantly different (p<0.05) in hours two and four between control group and tramadol. In all scales used there were significant difference (p<0.05) in hours 8, 10 and 12. In these times the morphine produced more quality analgesia than the tramadol. Based on the results obtained, it can be concluded that the tramadol administered in epidural route resulted a satisfactory analgesia, without an adverse effects, but poorer than the morphine analgesia administered by the same route. / O objetivo do presente estudo foi comparar a efic?cia e dura??o analg?sica do tramadol ?s da morfina, administrados por via epidural em gatos (Felis catus domesticus). Para tanto, utilizaram-se seis f?meas da esp?cie felina, sem ra?a definida, com peso entre tr?s a quatro quilos e idade variando de um a tr?s anos. Ap?s o jejum alimentar, de 12 horas e h?drico, de quatro horas, os animais foram submetidos ? anestesia geral com isoflurano por meio de uma c?mara anest?sica, com a finalidade de realizar a t?cnica epidural, evitando o estresse. Cada animal recebeu de forma aleat?ria, em um estudo cego, tr?s tratamentos por via epidural com intervalos de uma semana entre estes, com: solu??o salina a 0,9% num volume equivalente a 0,22ml.kg-1 (Grupo Controle), tramadol 1mg.kg-1 dilu?do em solu??o salina e volume equivalente a 0,22ml.kg-1 (Grupo Tramadol) e morfina 0,1mg.kg-1 dilu?do em solu??o salina e volume equivalente a 0,22ml.kg-1 (Grupo Morfina). Uma hora ap?s a administra??o de cada f?rmaco e o completo retorno da anestesia geral, os animais foram submetidos a est?mulos dolorosos em tr?s locais pr?-definidos: base da cauda, face lateral da coxa direita e esquerda, dando continuidade, 2, 3, 4, 6, 8, 10 e 12 horas respectivamente. A dor foi qualificada pela implementa??o de duas Escalas Simples Descritivas (SDS), uma Escala Anal?gica Visual (VAS), mensura??o da freq??ncia card?aca e respirat?ria. Para as vari?veis qualitativas utilizou-se o delineamento em Quadrado Latino 3 x 3 com o teste de Kruskal-Wallis e para as quantitativas a An?lise de Vari?ncias (ANOVA). N?o houve diferen?a em rela??o ao tempo de anestesia e freq??ncia respirat?ria, entretanto, houve diferen?a estat?stica significante (p<0,05) na freq??ncia card?aca nas horas dois e quatro entre os animais do grupo controle e do tramadol. Em todas as escalas utilizadas houve diferen?a significante (p<0,05) nas horas 8, 10 e 12, sendo que a morfina apresentou qualidade analg?sica superior ao do tramadol nesses momentos. Com base nos resultados obtidos, pode-se concluir que o tramadol administrado por via epidural resultou analgesia satisfat?ria, isenta de efeitos adversos, por?m inferior ? analgesia da morfina, administrada pela mesma via.

The pathophysiology of chronic morphine on the immune system, especially on the cells of the innate immune system that leads to an immune compromise state has not been fully elucidated. The cells of the innate immune system are the first line of defense in mounting an immune response needed in infections, inflammation, cancer development, etc. One of the ways by which these innate immune cells act is by the production of cytokines with direct effects and to also recruit other immune cells, as required. A balance of pro- and anti-inflammatory cytokines is necessary for immune competence. I hypothesized that chronic morphine would act via a classical opioid receptor to stimulate the PI3K/Akt/Gsk3β pathway to produce predominantly anti-inflammatory cytokines. Cytokine gene expression levels were assessed via RT-PCR; Akt and Gsk3β protein levels measured using indirect ELISA. The data suggests that chronic morphine causes a significant reduction in IL-6 production, but does not act via the Akt/Gsk3β pathway or the classical opioid receptor to cause this effect in microglia cells.

Protein engineering has been a useful tool in the fight against human diseases. Human insulin was the first recombinant DNA-derived therapeutic protein (Humulin®) approved by the US FDA in 1982. However, many of the early protein drugs were only recombinant versions of natural proteins with no modification of their primary amino acid sequence and most of them did not make optimal drug products mainly due to their short half-life or suboptimal affinity, leading to poor therapeutic efficacy. The difficulty in the large-scale production of some therapeutic proteins was another important issue. In the past three decades, different protein engineering platforms have been developed to overcome the obstacles seen in the first generations of these treatments. With the help of these new techniques, proteins have been purposefully modified to improve their clinical potential. The focus of my dissertation is the engineering of potential protein drugs to make them therapeutically useful and more valuable. Previously, our research group has developed cocaine hydrolases (CocHs) from human butyrylcholinesterase (BChE) for treatment of cocaine addiction and prevention of acute cocaine intoxication. In the first project, CocHs were further engineered to improve their performance, e.g., Fc-fused CocHs with an extended serum half-life. Then, I investigated the potential application of a long-lasting CocH for protection against the acute toxic and stimulant effects of cocaine. In the second project, I investigated the potential inhibition of CocH-mediated cocaine hydrolysis by heroin (3,6-diacetylmorphine) or its initial host metabolite, 6-monoacetylmorphine (6-MAM). The investigation of this possible inhibition was important to determine the in vivo efficacy of CocHs, as heroin is one of the most commonly co-abused drugs by cocaine-dependent individuals, as well as a possible metabolite of CocHs. In the third project, I expressed and characterized the recombinant human UDP-glucuronosyltransferase 1A10 (UGT1A10) enzyme, which can inactivate many therapeutically valuable substances. In the fourth and final project, prostate apoptosis response-4 (Par-4), a tumor suppressor protein, was engineered to have a prolonged duration of action so that it may be more therapeutically valuable for cancer treatment.