Submitted by DENISE MOROTO (de_moroto@hotmail.com) on 2018-04-18T15:30:46Z No. of bitstreams: 1 Tesev6.pdf: 9989693 bytes, checksum: b1f5424e844b4857860140c0b71b3cca (MD5) / Rejected by Luciana Pizzani null (luciana@btu.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: Necessário fazer as seguintes correções no arquivo submetido: problema 1: o arquivo submetido não contém capa padrão, item obrigatório de acordo com as normas do seu programa. Assim que tiver efetuado as correções submeta o arquivo em PDF novamente. Agradecemos a compreensão. on 2018-04-19T13:02:09Z (GMT) / Submitted by DENISE MOROTO (de_moroto@hotmail.com) on 2018-04-19T14:03:21Z No. of bitstreams: 1 Tesev7.pdf: 10060555 bytes, checksum: bf4975d0b77a02d79ca9201cb11a74b8 (MD5) / Approved for entry into archive by Luciana Pizzani null (luciana@btu.unesp.br) on 2018-04-19T14:53:49Z (GMT) No. of bitstreams: 1 moroto_d_dr_bot.pdf: 10060555 bytes, checksum: bf4975d0b77a02d79ca9201cb11a74b8 (MD5) / Made available in DSpace on 2018-04-19T14:53:49Z (GMT). No. of bitstreams: 1 moroto_d_dr_bot.pdf: 10060555 bytes, checksum: bf4975d0b77a02d79ca9201cb11a74b8 (MD5) Previous issue date: 2018-02-21 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Introdução: Dor perioperatória desencadeia resposta neuroendócrina ao estresse, responsável por efeitos adversos em vários órgãos e sistemas. Estratégias foram desenvolvidas para controle da dor e dessa resposta ao trauma, como a injeção de dexametasona, glicocorticoide de alta potência, no espaço peridural e subaracnoideo. Embora a injeção de corticosteroides no neuroeixo seja realizada com aparentes benefícios em seres humanos no contexto da dor aguda e crônica, a eficácia e segurança desse procedimento continuam discutíveis e alvo de controvérsia na literatura. O objetivo deste estudo foi avaliar os efeitos histológicos que a dexametasona administrada no espaço subaracnoideo determina sobre o tecido nervoso da medula espinal e das meninges dos coelhos. Método: Após aprovação pela Comissão de Ética no Uso de Animais, 28 coelhos adultos jovens foram randomizados em dois grupos (G) experimentais com 14 animais cada: G1 recebeu solução salina a 0,9% pela via subaracnoidea e G2, dexametasona na dose de 0,37 mg.kg-1. Os animais foram mantidos em cativeiro por 21 dias, durante os quais se avaliou motricidade e sensibilidade dolorosa. Depois desse período, sob anestesia venosa, foram submetidos a sacrifício para retirada da porção lombar e sacral da medula espinal e das meninges para avaliação histológica pelo método de Hematoxilina-eosina (HE) e de marcação imuno-histoquímica para Proteína Glial Fibrilar Ácida (GFAP). Resultados: Todos os animais permaneceram sem alterações clínicas durante o período de cativeiro. Nenhum animal do G1 apresentou alterações histológicas à microscopia óptica. No G2, 13 animais exibiram infiltrado inflamatório linfoplasmocitário perivascular nos vasos das meninges, com ou sem acometimento também das meninges e do parênquima nervoso. Os animais do G2 também tiveram percentual de células marcadas pelo GFAP inferior aos do G1 (p<0,05). Conclusão: Neste modelo experimental em coelhos, a solução de dexametasona determinou alterações histológicas no tecido nervoso da medula espinal e, principalmente, nas meninges. / Background: Perioperative pain triggers neuroendocrine response to stress, responsible for adverse effects on various organs and systems. Strategies were developed to control pain and this response to trauma, such as the injection of dexamethasone, a high-potency glucocorticoid, into the epidural and subarachnoid space. Although corticosteroid spinal injection is performed with apparent benefits in humans in the context of acute and chronic pain, the efficacy and safety of this procedure remain controversial in the literature. The objective of this study was to evaluate the histological effects that dexamethasone administered into the subarachnoid space determines on the nervous tissue of the spinal cord and the meninges of rabbits. Methods: After approval by the animal research ethics committee, 28 young adult rabbits were randomized in two experimental groups (G) with 14 animals each: G1 received 0.9% saline solution into the intrathecal space and G2 received 0.37mg.kg-1 of dexamethasone by the same route. The animals were clinically evaluated for 21 days. After this period of observation, sacrifice was performed under intravenous anesthesia and then the lumbar and sacral portion of their spinal cords were removed for histological examination by Hematoxylin-eosin (HE) and Glial Fibrillary Acidic protein (GFAP) immunohistochemical staining. Results: All animals remained without clinical changes during the period of captivity. No histological changes were observed in G1 animals. In G2, 13 animals exhibited perivascular lymphoplasmocytic inflammatory infiltrate in the meninges vessels, with or without involvement of the meninges and the nervous parenchyma. G2 animals also showed lower percentage of GFAP stained cells than rabbits of G1 (p<0.05). Conclusion: In this experimental model, the dexamethasone solution determined histological changes in the nervous tissue of the spinal cord and, especially, the meninges of rabbits. / FAPESP: 2014/23740-2

Orientador: Eliana Marisa Ganem / Resumo: Introdução: Dor perioperatória desencadeia resposta neuroendócrina ao estresse, responsável por efeitos adversos em vários órgãos e sistemas. Estratégias foram desenvolvidas para controle da dor e dessa resposta ao trauma, como a injeção de dexametasona, glicocorticoide de alta potência, no espaço peridural e subaracnoideo. Embora a injeção de corticosteroides no neuroeixo seja realizada com aparentes benefícios em seres humanos no contexto da dor aguda e crônica, a eficácia e segurança desse procedimento continuam discutíveis e alvo de controvérsia na literatura. O objetivo deste estudo foi avaliar os efeitos histológicos que a dexametasona administrada no espaço subaracnoideo determina sobre o tecido nervoso da medula espinal e das meninges dos coelhos. Método: Após aprovação pela Comissão de Ética no Uso de Animais, 28 coelhos adultos jovens foram randomizados em dois grupos (G) experimentais com 14 animais cada: G1 recebeu solução salina a 0,9% pela via subaracnoidea e G2, dexametasona na dose de 0,37 mg.kg-1. Os animais foram mantidos em cativeiro por 21 dias, durante os quais se avaliou motricidade e sensibilidade dolorosa. Depois desse período, sob anestesia venosa, foram submetidos a sacrifício para retirada da porção lombar e sacral da medula espinal e das meninges para avaliação histológica pelo método de Hematoxilina-eosina (HE) e de marcação imuno-histoquímica para Proteína Glial Fibrilar Ácida (GFAP). Resultados: Todos os animais permaneceram sem alterações clíni... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Background: Perioperative pain triggers neuroendocrine response to stress, responsible for adverse effects on various organs and systems. Strategies were developed to control pain and this response to trauma, such as the injection of dexamethasone, a high-potency glucocorticoid, into the epidural and subarachnoid space. Although corticosteroid spinal injection is performed with apparent benefits in humans in the context of acute and chronic pain, the efficacy and safety of this procedure remain controversial in the literature. The objective of this study was to evaluate the histological effects that dexamethasone administered into the subarachnoid space determines on the nervous tissue of the spinal cord and the meninges of rabbits. Methods: After approval by the animal research ethics committee, 28 young adult rabbits were randomized in two experimental groups (G) with 14 animals each: G1 received 0.9% saline solution into the intrathecal space and G2 received 0.37mg.kg-1 of dexamethasone by the same route. The animals were clinically evaluated for 21 days. After this period of observation, sacrifice was performed under intravenous anesthesia and then the lumbar and sacral portion of their spinal cords were removed for histological examination by Hematoxylin-eosin (HE) and Glial Fibrillary Acidic protein (GFAP) immunohistochemical staining. Results: All animals remained without clinical changes during the period of captivity. No histological changes were observed in G1 anima... (Complete abstract click electronic access below) / Doutor

Ultrasound-guided selective C5 nerve root blocks have been described in several case reports as a safe and effective means to anesthetize the distal clavicle while maintaining innervation of the upper extremity and preserving diaphragmatic function. In this study, cadavers were injected with 5 mL of 0.5% methylene blue dye under ultrasound guidance to investigate possible proximal and distal spread of injectate along the brachial plexus, if any. Following the injections, the specimens were dissected and examined to determine the distribution of dye and the structures affected. One injection revealed dye extended proximally into the epidural space, which penetrated the dura mater and was present on the spinal cord and brainstem. Dye was noted distally to the divisions in 3 injections. The anterior scalene muscle and phrenic nerve were stained in all 4 injections. It appears unlikely that local anesthetic spread is limited to the nerve root following an ultrasound-guided selective C5 nerve root injection. Under certain conditions, intrathecal spread also appears possible, which has major patient safety implications. Additional safety measures, such as injection pressure monitoring, should be incorporated into this block, or approaches that are more distal should be considered for the acute pain management of distal clavicle fractures.

Le projet porte sur l’étude de l’effet de l’eugénol, composant principal du clou de girofle, sur la douleur neuropathique. L’objectif principal du projet était de déterminer la contribution du système nerveux central dans l’effet analgésique de l’eugénol. Lors d’une étude préliminaire, la pénétrabilité de l’eugénol a été évaluée dans le système nerveux central du rat. Des échantillons de sang, de cerveau et de moelle épinière ont été prélevés et les concentrations d’eugénol dans ces différents tissus ont été analysées à l’aide d’un spectromètre de masse. Les résultats ont montré que l’eugénol pénètre bien le système nerveux central avec une distribution plus importante dans la moelle épinière. Après l’induction de la douleur neuropathique à des rats Sprague-Dawley par le modèle de ligatures du nerf sciatique, des injections intrathécales d’eugénol furent réalisées afin d’évaluer l’effet central de l’eugénol. La plus forte dose d’eugénol a atténué l’allodynie secondaire après 15min, 2h et 4h et a aussi amélioré l’hyperalgésie thermique après 2h et 4h. Ces résultats confirment l’hypothèse que l’eugénol atténue les deux aspects de la douleur neuropathique que sont l’allodynie et l’hyperalgésie. Les injections au niveau lombaire permettent de penser que l’eugénol, un agoniste/antagoniste des récepteurs vanilloïdes pourrait diminuer la douleur neuropathique en agissant notamment au niveau des récepteurs vanilloïdes situés dans la corne dorsale de la moelle épinière. / The project is based on the study of the effects of eugenol, the main component of clove oil, on neuropathic pain. The main objective was to determine the central effect of eugenol. In a preliminary study we evaluated the penetrability of eugenol in the central nervous system of rats. Blood, brain and spinal cord samples were collected and concentrations were determined by mass spectrometry. Brain-toplasma and spinal cord-to-plasma ratios suggest that eugenol penetrates the central nervous system of rats relatively well, with a preferential distribution in the spinal cord. Following the induction of neuropathic pain in male Sprague-Dawley rats using the sciatic nerve ligation model, intrathecal injections of eugenol were done to evaluate the central effect of eugenol. Treatment with the high dose of eugenol significantly decreased secondary mechanical allodynia measured by the Von Frey test after 15min, 2h and 4h and improved thermal hyperalgesia measured by the Hargreaves device after 2h and 4h. Results support the hypothesis that eugenol may alleviate neuropathic pain, both allodynia and hyperalgesia. Eugenol, a vanilloid agonist/antagonist may therefore reduce neuropathic pain by acting on vanilloid receptors at the level of the dorsal horn of the spinal cord.

Le projet porte sur l’étude de l’effet de l’eugénol, composant principal du clou de girofle, sur la douleur neuropathique. L’objectif principal du projet était de déterminer la contribution du système nerveux central dans l’effet analgésique de l’eugénol. Lors d’une étude préliminaire, la pénétrabilité de l’eugénol a été évaluée dans le système nerveux central du rat. Des échantillons de sang, de cerveau et de moelle épinière ont été prélevés et les concentrations d’eugénol dans ces différents tissus ont été analysées à l’aide d’un spectromètre de masse. Les résultats ont montré que l’eugénol pénètre bien le système nerveux central avec une distribution plus importante dans la moelle épinière. Après l’induction de la douleur neuropathique à des rats Sprague-Dawley par le modèle de ligatures du nerf sciatique, des injections intrathécales d’eugénol furent réalisées afin d’évaluer l’effet central de l’eugénol. La plus forte dose d’eugénol a atténué l’allodynie secondaire après 15min, 2h et 4h et a aussi amélioré l’hyperalgésie thermique après 2h et 4h. Ces résultats confirment l’hypothèse que l’eugénol atténue les deux aspects de la douleur neuropathique que sont l’allodynie et l’hyperalgésie. Les injections au niveau lombaire permettent de penser que l’eugénol, un agoniste/antagoniste des récepteurs vanilloïdes pourrait diminuer la douleur neuropathique en agissant notamment au niveau des récepteurs vanilloïdes situés dans la corne dorsale de la moelle épinière. / The project is based on the study of the effects of eugenol, the main component of clove oil, on neuropathic pain. The main objective was to determine the central effect of eugenol. In a preliminary study we evaluated the penetrability of eugenol in the central nervous system of rats. Blood, brain and spinal cord samples were collected and concentrations were determined by mass spectrometry. Brain-toplasma and spinal cord-to-plasma ratios suggest that eugenol penetrates the central nervous system of rats relatively well, with a preferential distribution in the spinal cord. Following the induction of neuropathic pain in male Sprague-Dawley rats using the sciatic nerve ligation model, intrathecal injections of eugenol were done to evaluate the central effect of eugenol. Treatment with the high dose of eugenol significantly decreased secondary mechanical allodynia measured by the Von Frey test after 15min, 2h and 4h and improved thermal hyperalgesia measured by the Hargreaves device after 2h and 4h. Results support the hypothesis that eugenol may alleviate neuropathic pain, both allodynia and hyperalgesia. Eugenol, a vanilloid agonist/antagonist may therefore reduce neuropathic pain by acting on vanilloid receptors at the level of the dorsal horn of the spinal cord.

The fever response, besides being part of host defense response to infection or inflammation, is associated with discomfort and anxiety and may constitute a risk for febrile seizures in children. Therefore, antipyretic therapy is routinely prescribed for febrile patients. The animal models of fever using the systemic injection of lipopolysaccharide (LPS) and Baker yeast, described in the literature, are suitable for screening of novel antipyretics, but they do not provide information regarding the mechanism of action of these compounds. Therefore, the present study aimed to describe and validate a model of fever induction by prostaglandin (PG) E2, the final mediator of febrile response in the central nervous system, in young male Wistar rats (25-30 days of age). In this protocol, PGE2 was injected intrathecally without implantation of cannula. Rectal temperature (TR) was recorded every thirty minutes for three hours after PGE2 injection (08:00 11:00 h). The intrathecal (i.t.) injection of PGE2 10 ηg in 100 μL/animal induced fever in the animals, which was prevented by administration of EP1 and EP3 receptors antagonists, but did not by antagonist of EP4 receptor. In addition, the classic antipyretics dipyrone and acetaminophen, at doses that had no effect per se on TR of animals, did not revert the fever induced by i.t. injection of PGE2. This model seems suitable to investigate whether the action of antipyretics occurs upstream or downstream the prostaglandin coupling in EP receptors. In addition, this protocol is advantageous from the technical, ethical and economical point of view compared to others PGE2-induced fever protocols described in the literature, because trepanation for cannula implantation is not required, reducing the inflammatory response, animals suffering and experimental costs. / A febre, apesar de fazer parte da resposta de defesa do hospedeiro à infecção ou inflamação, está associada com desconforto e ansiedade, além de representar um risco iminente de convulsões febris em crianças. Por isso, terapia antipirética é rotineiramente prescrita a pacientes febris. Os modelos animais de febre empregando a injeção sistêmica de lipopolissacarídeo (LPS) e fermento de padeiro, descritos na literatura, são úteis para a triagem de novos antipiréticos, mas não fornecem informações a respeito do mecanismo de ação desses compostos. Diante disso, o presente estudo objetivou padronizar e validar um modelo de indução de febre por prostaglandina (PG) E2, o mediador final da resposta febril no sistema nervoso central, em ratos machos jovens da raça Wistar (25-30 dias). Neste protocolo, a PGE2 foi injetada pela via intratecal (i.t.), não necessitando a implantação de cânula. A temperatura retal (TR) foi registrada a cada trinta minutos durante três horas após a injeção da PGE2 (08:00-11:00 h). A injeção i.t. de PGE2 10 ηg em 100 μL/animal induziu febre nos animais, a qual foi prevenida pela administração de antagonistas dos receptores EP1 e EP3, mas não por antagonista do receptor EP4. Além disso, os antipiréticos clássicos dipirona e paracetamol, em doses que não tiveram efeito per se na TR dos animais, não reverteram a febre induzida por PGE2 i.t. Este modelo parece útil para investigar se a ação dos antipiréticos ocorre antes ou depois da ligação da PGE2 em seus receptores EP. Além disso, este protocolo é vantajoso do ponto de vista técnico, ético e econômico em relação aos outros protocolos de indução de febre por PGE2 descritos na literatura, porque a trepanação para implantação de cânula não é necessária, reduzindo a resposta inflamatória, o sofrimento dos animais e os custos experimentais.