Made available in DSpace on 2014-06-11T19:22:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-22Bitstream added on 2014-06-13T20:09:23Z : No. of bitstreams: 1 lopes_mcs_me_jabo.pdf: 1425980 bytes, checksum: ec77cfbfaaacc7792256be3564bbc255 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Avaliaram-se os efeitos clínico e comportamental da injeção intravenosa do tramadol no controle da dor induzida experimentalmente, por obstrução intestinal extraluminal, com dreno de Pen Rose. Foram utilizados 24 cavalos distribuídos em quatro grupos: controle (GC, n=6); obstrução duodenal (GD, n=6); obstrução de íleo (GI, n=6) e obstrução de flexura pélvica (GFP, n=6). Após medicação pré-anestésica com a associação de acepromazina (0,025 mg.kg-1 IV), xilazina (0,5 mg.kg-1 IV) e meperidina (4 mg.kg-1 IM), o tramadol foi administrado nas doses de 1,0 mg.kg-1 e 1,5 mg.kg-1, por via intravenosa (IV), imediatamente após a obstrução intestinal, em três cavalos de cada grupo. Avaliaram-se as freqüências cardíaca (FC) e respiratória (f), temperatura retal (TºC), tempo de preenchimento capilar (TPC), motilidade intestinal, comportamento relacionado à dor (olhar para o flanco, cavar, deitar e rolar) hemograma e hemogasometria venosa, nos intervalos: M0 (basal) a cada 0,5 hora de M1 a M6 , na fase de obstrução, e até três horas após a reversão do processo obstrutivo (M7 a M12). Os resultados demonstraram que não houve diferença significativa entres as doses utilizadas dentro de cada grupo, assim como entre os grupos. Houve aumento da FC em M11 no GD e em M12 no GFP. Os sinais de dor abdominal e atonia intestinal iniciaram-se em M5 no GFP e em M6 no GI. Nos animais do GD, os sinais de desconforto não progrediram. No leucograma foi observado um quadro característico de estresse e na hemogasometria os animais do GD tendenciaram à alcalose metabólica com compensação respiratória. Clinicamente, observou-se que a dose de 1,5 mg.kg-1 de tramadol proporcionou melhor conforto para os animais, porem sem significado estatístico, quando comparado coma dose de 1,0 mg.kg-1... / The clinical and behavioral effects of the intravenous injection of tramadol were evaluated during the control of pain induced experimentally due to intestinal extraluminal obstruction using “Pen Rose” drain. A total of 24 horses were used and distributed in four groups: control (GC, n=6); duodenal obstruction (GD, n=6); ileum obstruction (GI, n=6) and pelvic flexure obstruction (GFP, n=6). After administration of pre-anesthetic medications using association of acepromazine (0.025 mg.kg-1 IV), xylazine (0.5 mg.kg-1 IV) and meperidine (4 mg.kg-1 IM), tramadol was administered at doses of 1.0 mg.kg-1 and 1.5 mg.kg-1 intravenously (IV), immediately after the intestinal obstruction in three horses of each group. Evaluations were performed, including heart rate (HR), respiratory rate (RR), rectal temperature (RT), capillary refill time (CRT), gut motility, pain-related behaviour (look for the sidewall, dig down and roll) and blood gases from venous blood at the time: M0 (baseline) and every 0.5 hours from M1 to M6, during obstruction process and also until three hours after the obstructive process be reverted (M7 to M12). The results showed no significant difference among the doses used in the same group as among groups. There was an increase in HR in the GD M11 and M12 of GFP. Signs of abdominal pain and intestinal atony began at M5 in GFP and at M6 in GI. In animals from GD, the discomfort signs did not showed progress. On the leucogram was observed a typical stress and on the blood gas analysis the animals from GD showed a tendency to metabolic alkalosis with respiratory compensation. Clinically, was observed that the dose of 1.5 mg.kg-1 of tramadol provided better comfort to the animals, but there was not statistical significance, compared with the dose 1.0 mg.kg-1... (Complete abstract click electronic access below)

Orientador: Carlos Augusto Araújo Valadão / Banca: Juan Carlos Duque Moreno / Banca: Paulo Sérgio Patto dos Santos / Resumo: Avaliaram-se os efeitos clínico e comportamental da injeção intravenosa do tramadol no controle da dor induzida experimentalmente, por obstrução intestinal extraluminal, com dreno de Pen Rose. Foram utilizados 24 cavalos distribuídos em quatro grupos: controle (GC, n=6); obstrução duodenal (GD, n=6); obstrução de íleo (GI, n=6) e obstrução de flexura pélvica (GFP, n=6). Após medicação pré-anestésica com a associação de acepromazina (0,025 mg.kg-1 IV), xilazina (0,5 mg.kg-1 IV) e meperidina (4 mg.kg-1 IM), o tramadol foi administrado nas doses de 1,0 mg.kg-1 e 1,5 mg.kg-1, por via intravenosa (IV), imediatamente após a obstrução intestinal, em três cavalos de cada grupo. Avaliaram-se as freqüências cardíaca (FC) e respiratória (f), temperatura retal (TºC), tempo de preenchimento capilar (TPC), motilidade intestinal, comportamento relacionado à dor (olhar para o flanco, cavar, deitar e rolar) hemograma e hemogasometria venosa, nos intervalos: M0 (basal) a cada 0,5 hora de M1 a M6 , na fase de obstrução, e até três horas após a reversão do processo obstrutivo (M7 a M12). Os resultados demonstraram que não houve diferença significativa entres as doses utilizadas dentro de cada grupo, assim como entre os grupos. Houve aumento da FC em M11 no GD e em M12 no GFP. Os sinais de dor abdominal e atonia intestinal iniciaram-se em M5 no GFP e em M6 no GI. Nos animais do GD, os sinais de desconforto não progrediram. No leucograma foi observado um quadro característico de estresse e na hemogasometria os animais do GD tendenciaram à alcalose metabólica com compensação respiratória. Clinicamente, observou-se que a dose de 1,5 mg.kg-1 de tramadol proporcionou melhor conforto para os animais, porem sem significado estatístico, quando comparado coma dose de 1,0 mg.kg-1... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The clinical and behavioral effects of the intravenous injection of tramadol were evaluated during the control of pain induced experimentally due to intestinal extraluminal obstruction using "Pen Rose" drain. A total of 24 horses were used and distributed in four groups: control (GC, n=6); duodenal obstruction (GD, n=6); ileum obstruction (GI, n=6) and pelvic flexure obstruction (GFP, n=6). After administration of pre-anesthetic medications using association of acepromazine (0.025 mg.kg-1 IV), xylazine (0.5 mg.kg-1 IV) and meperidine (4 mg.kg-1 IM), tramadol was administered at doses of 1.0 mg.kg-1 and 1.5 mg.kg-1 intravenously (IV), immediately after the intestinal obstruction in three horses of each group. Evaluations were performed, including heart rate (HR), respiratory rate (RR), rectal temperature (RT), capillary refill time (CRT), gut motility, pain-related behaviour (look for the sidewall, dig down and roll) and blood gases from venous blood at the time: M0 (baseline) and every 0.5 hours from M1 to M6, during obstruction process and also until three hours after the obstructive process be reverted (M7 to M12). The results showed no significant difference among the doses used in the same group as among groups. There was an increase in HR in the GD M11 and M12 of GFP. Signs of abdominal pain and intestinal atony began at M5 in GFP and at M6 in GI. In animals from GD, the discomfort signs did not showed progress. On the leucogram was observed a typical stress and on the blood gas analysis the animals from GD showed a tendency to metabolic alkalosis with respiratory compensation. Clinically, was observed that the dose of 1.5 mg.kg-1 of tramadol provided better comfort to the animals, but there was not statistical significance, compared with the dose 1.0 mg.kg-1... (Complete abstract click electronic access below) / Mestre

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Protium heptaphyllum March (Burseraceae), a medicinal plant commonly found in the Amazon and in the Northeast regions of Brazil, releases an oil-resin rich in pentacyclic triterpenes, such as the binary mixture of alpha- and beta- amyrin, that manifests antiinflamatory, antinociceptive and gastroprotective properties. This work was aimed to evaluate the antinociceptive effect of the alpha- and beta- amyrin mixture in the cyclophosphamide (400 mg/kg), acetic acid (0,6%, 10mL/kg, i.p.) and mustard oil-induced visceral nociception models in mice and to establish the likely mechanism(s) of action. In the cyclophosphamide-induced visceral pain model, pretreatment of mice with triterpene mixture at the oral doses of 10, 30 and 100 mg/kg significantly reduced (p<0.001) the pain-related behavioral expression time (59,7; 75,5 e 92,3%, respectively, versus the cyclophosphamide-treated group 12,25 +/- 2,98 min) in a dose-dependent manner. Suppression of visceral painârelated behaviors was also evidenced to the triterpenoid mixture (10 mg/kg) in the intraperitoneal acetic acid- and intracolonically injected mustard oil-induced test models of visceral nociception 50,4% e 61,1%, respectively compared to the acetic acid-treated group (42,33 +/- 3,78 abdominal constrictions/20 min) in the writhing test and to the control in the mustard oil (0,75%, 50 mcL/animal) experiment (39,28 +/- 3,26). In these tests, the maximal suppression of visceral pain was observed at 10 mg/kg. The possible mechanisms involved in the antinociceptive action of alpha- and beta- amyrin (10 mg/kg) were analyzed in the mustard oil-induced visceral pain model. In the evaluation of the opioid receptor involvement, both the triterpene mixture and morphine (5 mg/kg, s.c.) effectively inhibited (p<0.001) the number of pain-related behaviors, which could be significantly reversed by pretreatment of animals with an opioid antagonist naloxona (2mg/kg, i.p.), suggesting the opioid participation in the alpha- and beta- amyrin mechanism of action. In the study of the alpha2-adrenoreceptor involvement, the triterpene mixture as well as clonidine (0.1 mg/kg, i.p.), a known alpha2 agonist, inhibited (p<0.001) the nociceptive behavioral expression. However, when the animals were pretreated with yohimbine, an alpha2-adrenoreceptor antagonist, only the inhibitory action of clonidine was reversed, suggesting the non-participation of alpha2- adrenoreceptor in the antinociceptive action of alpha- and beta- amyrin. In the evaluation of TRPV1 receptor involvement, mice pretreated with either the alpha- and beta- amyrin, ruthenium red, a TRPV1 non-competitive antagonist, (3 mg/kg, s.c.) or their combination induced a significant and similar inhibition (p<0.001) of the number of nociceptive behaviors. The degree of inhibition with no potentiation or antagonism suggests that alpha- and beta- amyrin may act as a TRPV1 non-competitive antagonist, like ruthenium red. In order to evaluate a possible sedative, motor impairment and motor incoordination effects related to alpha- and beta- amyrin, the penthobarbitone-induced sleeping time, open-field and rota-rod tests were performanced, respectively. The data indicated that the treatment of animals with the alpha- and beta- amyrin mixture (10 mg/kg) was unable to cause sedation, motor impairment or motor incoordination effects (p>0.05), being even able to reverse (p<0.05) a mustard oil-induced motor impairment in the open field test. The results taken together strongly suggest the therapeutic potential of alpha- and beta- amyrin in oblitering visceral nociception through the mechanisms that involve the opioids and TRPV1 receptors. / O Protium heptaphyllum March. (Burseraceae), uma planta medicinal encontrada na regiÃo AmazÃnica e Nordeste do Brasil, produz uma resina rica em triterpenos pentacÃclicos, como a mistura binÃria alpha- e beta- amirina, que apresentam atividade antiinflamatÃria, gastroprotetora e antinociceptiva. Este trabalho objetivou investigar a atividade antinociceptiva de alpha- e beta- amirina em modelos de dor visceral induzida por ciclofosfamida, Ãcido acÃtico e Ãleo de mostrada em camundongos, alÃm dos possÃveis mecanismos de aÃÃo envolvidos. No modelo de nocicepÃÃo visceral induzida por ciclofosfamida (400 mg/kg, i.p.), a mistura de triterpenos nas doses de 10, 30 e 100 mg/kg, v.o., reduziu (p<0,001) de forma dose-dependente o tempo de expressÃo dos comportamentos relacionados à dor visceral (59,7; 75,5 e 92,3%, respectivamente, versus o controle ciclofosfamida 12,25 +/- 2,98 min). Realizou-se o estudo nos modelos de contorÃÃes abdominais induzidas por Ãcido acÃtico (0,6%, 10mL/kg, i.p.) e dor visceral induzida por Ãleo de mostarda (0,75%, 50 mcL/animal) intracolÃnico. Os resultados indicaram uma inibiÃÃo do nÃmero de comportamentos de dor expressos pelos animais, sendo o maior nÃvel de inibiÃÃo (p<0,001) encontrado na dose de 10 mg/kg da alpha- e beta- amirina 50,4% e 61,1% comparados respectivamente ao controle Ãcido acÃtico (42,33 +/- 3,78 contorÃÃes/20min) no teste de contorÃÃes abdominais e ao controle Ãleo de mostarda (39,28 +/- 3,26) no modelo de dor visceral por Ãleo de mostarda. Para o estudo do possÃvel mecanismo de aÃÃo de alpha- e beta- amirina foi utilizada a dose de 10 mg/kg da mistura de triterpenos no modelo de nocicepÃÃo por Ãleo de mostarda. Na avaliaÃÃo da participaÃÃo do sistema opiÃide, a mistura dos triterpenos e a morfina (5 mg/kg, s.c.) inibiram significativamente (p<0,001) o nÃmero de comportamentos de dor expressos, havendo uma reversÃo da antinocicepÃÃo (p<0,05) quando prÃ-tratados com naloxona (2 mg/kg, i.p.), sugerindo a participaÃÃo opiÃide no mecanismo da alpha- e beta- amirina. No estudo do envolvimento do sistema adrenÃrgico, a mistura de triterpenos e a clonidina (0,1 mg/kg, i.p.), um agonista alpha2-adrenÃrgico, inibiram (p<0,001) a expressÃo dos comportamentos nociceptivos. PorÃm, com o prÃ-tratamento com ioimbina, um antagonista alpha2, houve reversÃo (p<0,05) da antinocicepÃÃo induzida pela clonidina, mas nÃo da alpha- e beta- amirina, sugerindo o nÃo envolvimento deste receptor na antinocicepÃÃo da mistura de triterpenos. No estudo do envolvimento do receptor TRPV1, o prÃ-tratamento dos animais com alpha- e beta- amirina, vermelho de rutÃnio (3 mg/kg, s.c.), um antagonista nÃo competitivo deste receptor, ou com a combinaÃÃo da mistura de triterpenos com vermelho de rutÃnio, houve uma inibiÃÃo (p<0,001) semelhante, para todos os tratamentos, dos comportamentos de dor. A nÃo potencializaÃÃo, ou antagonismo, do efeito antinociceptivo de alpha- e beta- amirina pelo vermelho de rutÃnio sugere que a mistura atue como um antagonista nÃo-competitivo TRPV1. Para avaliar a existÃncia de um efeito sedativo, de um impedimento locomotor ou de uma incoordenaÃÃo motora, foram utilizados os testes do tempo de sono induzido por pentobarbital, teste do campo aberto e o teste do rota rod, respectivamente. Os dados indicaram que o tratamento com a mistura de triterpenos (10 mg/kg) nÃo induziu (p>0,05) sedaÃÃo, impedimento locomotor ou incoordenaÃÃo motora nos animais, sendo ainda capaz de reverter (p<0,05) o impedimento locomotor induzido pelo Ãleo de mostarda no teste do campo aberto. Em conjunto os dados revelaram a efetividade da mistura de alpha- e beta- amirina em modelos de nocicepÃÃo visceral possivelmente envolvendo receptores opiÃides e TRPV1.

ROCHA, MG. Mediadores inflamatórios na dor pélvica crônica Identificação de possíveis marcadores séricos da doença. Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, 2010. Introdução: Dor pélvica crônica é uma doença de elevada prevalência e fisiopatologia complexa. Os métodos diagnósticos muitas vezes são insuficientes e, em decorrência, o tratamento e seguimento das mulheres é difícil. Inúmeras doenças que se apresentam com dor crônica tem um perfil inflamatório, que ainda não foi investigado para o tema em questão. Objetivos: Quantificar os níveis de óxido nítrico (NO) e das metaloproteinases 2 (MMP-2) e 9 (MMP-9) no plasma de mulheres com dor pélvica crônica. Pacientes e métodos: Foram incluídas 64 mulheres, subdivididas em 02 grupos: dor pélvica crônica e grupo controle, com 37 pacientes no primeiro grupo e 27 pacientes no segundo grupo. As pacientes do grupo de estudo eram seguidas no Ambulatório de Dor Pélvica e Endoscopia e as pacientes do grupo controle eram seguidas no Ambulatório de Anticoncepção do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto USP. Foi realizada a mensuração clínica da dor através de uma escala unidimensional (VAS) e uma escala multidimensional (McGill). Também foram preenchidas as escalas de ansiedade e depressão (HAD). Indivíduos com qualquer evidência de processos inflamatórios, hipertensão, tabagismo ou uso de contraceptivos hormonais foram excluídos. Indivíduos tomando medicação para a dor, como analgésicos ou antiinflamatórios foram solicitados a pará-los 72 horas antes de participar do estudo. Foi coletada uma amostra sanguínea de 10 ml, no ato da consulta. Esse material foi armazenado em frasco próprio com anti-coagulante, processado imediatamente no local para separação do plasma e armazenado em freezer, a -70C para mensuração subseqüente. As concentrações de espécies relacionadas ao NO (nitrato) em líquidos foram medidas, sempre em triplicata, pelo método da quimioluminescência, que é um dos métodos mais simples, sensíveis e precisos disponíveis para medir NO. Foi utilizado um analisador de NO (Sievers Model 280 NO Analyzer - Boulder, CO, EUA), o qual permite medir NO em quantidades tão pequenas quanto 1 pmol. A atividade das MMP-2 e MMP-9 no plasma serão determinadas pelo método da zimografia, que consiste em uma eletroforese das amostras em um sistema SDS/PAGE que inclui o substrato da enzima (gelatina) no gel de separação, de modo a permitir a evidenciação e quantificação da atividade da enzima. Resultados: Os níveis plasmáticos de NO foram maiores nas pacientes com DPC quando comparadas às pacientes do grupo controle (16.8 ± 7.9 versus 12.2 ± 2.4, respectivamente (P = 0.0016). Especificamente, os níveis plasmáticos de NO foram maiores nas pacientes com DPC de origem visceral quando comparadas às pacientes com dor exclusivamente somática ou aos controles saudáveis (19.2 ± 8.9 versus 12.4 ± 1.8 versus 12.2 ± 2.4, respectivamente) (P=0.0001). Não observamos uma correlação entre os níveis plasmáticos de NO e a duração dos sintomas (em meses) (Spearman r = 0.04, 95%CI:-0.34 to 0.40, P = 0.84) ou com a intensidade dos sintomas dolorosos: EAV (Spearman r =:-0.18, 95%CI:-0.52 to 0.20, P=0.34), ou McGill (Spearman r = -0.06, 95%CI:-0.41 to 0.30, P =0.72). Com relação às MMP´s, não houve diferença estatística entre os dois grupos. Conclusões: Os níveis plasmáticos de NO encontram-se elevados em mulheres com DPC, especialmente naquelas com dor de origem visceral. Este fato pode ser considerado uma possibilidade no seguimento de pacientes com DPC, visto que pode ser usado como um marcador sérico para a doença. Já a dosagem das MMP-s não se mostrou útil como marcador plasmático para mulheres com DPC. / Background: Chronic pelvic pain is a disease of high prevalence and a complex pathophysiology. The diagnostic methods are often inadequate and, consequently, treatment and follow-up of women is quite difficult. Several diseases that present with chronic pain has an inflammatory profile, which has not yet been investigated for the topic. Aim: to quantify levels of nitric oxide (NO) and metalloproteinases 2 (MMP-2) and 9 (MMP-9) in plasma of women with chronic pelvic pain. Methods: 64 women were included in the sudy and divided into 02 groups: chronic pelvic pain and control group with 37 patients in the first group and 27 patients in the second group. Patients in the study group were followed at the Endoscopy and Pelvic Pain Unit and the control group patients were followed in the Contraception Unit of the Hospital of the Medical School of Ribeirão Preto University of São Paulo. We performed the measurement of clinical pain by a unidimensional scale (VAS) and a multidimensional scale (McGill). Anxiety and depression scales were also filled. Individuals with any evidence of inflammation, hypertension, smoking or use of hormonal contraceptives were excluded. Individuals taking medication for pain, such as painkillers or antiinflammatory drugs were asked to stop them 72 hours before entering the study. A blood sample was collected from 10 ml during the appointment. This material was stored in bottle itself with anti-coagulant (EDTA and / or heparin), processed immediately on site for plasma separation and stored in a -70 ºC freezer. The concentrations of species related to NO were measured in liquid, always in triplicate by the method of chemiluminescence, which is one of the most simple, sensitive and accurate available to measure NO. We used a NO analyzer (Sievers Model 280 NO Analyzer - Boulder, CO, USA), which allows the measurement of NO in quantities as small as 1 pmol. The activity of MMP-2 and MMP-9 in plasma was determined by the zymography method, which consists of an electrophoresis of the samples in an SDS / PAGE system, which includes the enzyme substrate (gelatin) in the gel separation, allowing the disclosure and quantification of enzyme activity. Results: Plasma NO levels were higher in CPP women than in controls (16.8 ± 7.9 versus 12.2 ± 2.4, respectively) (P=0.0016). Furthermore, plasma nitrate levels were higher in CPP women with evidence of pain of visceral origin than in CPP women with evidence of an exclusive somatic component or controls (19.2 ± 8.9 versus 12.4 ± 1.8 versus 12.2 ± 2.4, respectively) (P=0.0001). No correlation was detected between NO levels and duration of symptoms or intensity of pain. Regarding the MMP\'s, there was no statistical difference between the two groups. Conclusion: Plasma levels of NO are elevated in women with CPP, especially those with visceral pain. This fact can be considered an option in treating patients with CPP as it can be used as a serum marker for the disease. On the other hand, MMP´s did not turn out to be a good serum marker for women with CPP.

Background: Altered autonomic nervous system (ANS) function has been proposed as a mechanism in the development of central sensitisation (CS) and visceral pain hypersensitivity (VPH). The contribution of the parasympathetic nervous system (PNS) and the factors that mediate differences in sensitisation to acid are unclear and their study will clarify risk factors for oesophageal pain hypersensitivity (OPH) in gastrooesophageal reflux disease. Aims: To investigate psychophysiological and pharmacological manipulation of PNS tone in the development of OPH, and to determine factors which predict the development of OPH to acid infusion in healthy volunteers in a validated model of acid induced OPH. Methods: Pain thresholds to electrical stimulation in the proximal oesophagus were determined before and after a 30-minute distal oesophageal infusion of 0.15 mol/L hydrochloric acid in subjects. Sympathetic (SNS) and PNS parameters were measured at baseline and continuously thereafter. Subjects underwent psychological profiling for anxiety, depression, attachment vulnerability and personality type. Using this model, five studies were undertaken: Study 1 a pilot study to trail modulation suitability for further study used. In Study 2, subjects who demonstrated secondary hyperalgesia in the proximal non-acid-exposed oesophagus performed deep or sham breathing. Study 3 subjects, who did not sensitise to acid, underwent a validated stress test to induce OPH. With Study 4, deep breathing with IV saline (placebo) or atropine (PNS antagonist) was used to evaluate deep breathing’s induced PNS tone in OPH reduction. Study 5, a genetic pilot study, exploring the role of the GCH-1 haplotype in VPH. Results: ANS control’s key role in CS was clarified. Deep breathing increased PNS tone and prevented acid-induced OPH in comparison to sham breathing and confirmed increased PNS tone’s reversal of OPH. Psychological factors of anxiety, alexithymia and attachment status influence ANS modulation of CS. Individuals’ predisposition to VPH due to psychogenetic profiles were clarified and their biopsychosocial role illustrated. Conclusions and Inferences: A mechanistic explanation for the analgesic effect of deep breathing is provided with potential therapeutic implications in the treatment of VPH syndromes. Further clinical study is warranted to develop cost-effective treatments for chronic VPH syndromes.

ROCHA, MG. Mediadores inflamatórios na dor pélvica crônica Identificação de possíveis marcadores séricos da doença. Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, 2010. Introdução: Dor pélvica crônica é uma doença de elevada prevalência e fisiopatologia complexa. Os métodos diagnósticos muitas vezes são insuficientes e, em decorrência, o tratamento e seguimento das mulheres é difícil. Inúmeras doenças que se apresentam com dor crônica tem um perfil inflamatório, que ainda não foi investigado para o tema em questão. Objetivos: Quantificar os níveis de óxido nítrico (NO) e das metaloproteinases 2 (MMP-2) e 9 (MMP-9) no plasma de mulheres com dor pélvica crônica. Pacientes e métodos: Foram incluídas 64 mulheres, subdivididas em 02 grupos: dor pélvica crônica e grupo controle, com 37 pacientes no primeiro grupo e 27 pacientes no segundo grupo. As pacientes do grupo de estudo eram seguidas no Ambulatório de Dor Pélvica e Endoscopia e as pacientes do grupo controle eram seguidas no Ambulatório de Anticoncepção do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto USP. Foi realizada a mensuração clínica da dor através de uma escala unidimensional (VAS) e uma escala multidimensional (McGill). Também foram preenchidas as escalas de ansiedade e depressão (HAD). Indivíduos com qualquer evidência de processos inflamatórios, hipertensão, tabagismo ou uso de contraceptivos hormonais foram excluídos. Indivíduos tomando medicação para a dor, como analgésicos ou antiinflamatórios foram solicitados a pará-los 72 horas antes de participar do estudo. Foi coletada uma amostra sanguínea de 10 ml, no ato da consulta. Esse material foi armazenado em frasco próprio com anti-coagulante, processado imediatamente no local para separação do plasma e armazenado em freezer, a -70C para mensuração subseqüente. As concentrações de espécies relacionadas ao NO (nitrato) em líquidos foram medidas, sempre em triplicata, pelo método da quimioluminescência, que é um dos métodos mais simples, sensíveis e precisos disponíveis para medir NO. Foi utilizado um analisador de NO (Sievers Model 280 NO Analyzer - Boulder, CO, EUA), o qual permite medir NO em quantidades tão pequenas quanto 1 pmol. A atividade das MMP-2 e MMP-9 no plasma serão determinadas pelo método da zimografia, que consiste em uma eletroforese das amostras em um sistema SDS/PAGE que inclui o substrato da enzima (gelatina) no gel de separação, de modo a permitir a evidenciação e quantificação da atividade da enzima. Resultados: Os níveis plasmáticos de NO foram maiores nas pacientes com DPC quando comparadas às pacientes do grupo controle (16.8 ± 7.9 versus 12.2 ± 2.4, respectivamente (P = 0.0016). Especificamente, os níveis plasmáticos de NO foram maiores nas pacientes com DPC de origem visceral quando comparadas às pacientes com dor exclusivamente somática ou aos controles saudáveis (19.2 ± 8.9 versus 12.4 ± 1.8 versus 12.2 ± 2.4, respectivamente) (P=0.0001). Não observamos uma correlação entre os níveis plasmáticos de NO e a duração dos sintomas (em meses) (Spearman r = 0.04, 95%CI:-0.34 to 0.40, P = 0.84) ou com a intensidade dos sintomas dolorosos: EAV (Spearman r =:-0.18, 95%CI:-0.52 to 0.20, P=0.34), ou McGill (Spearman r = -0.06, 95%CI:-0.41 to 0.30, P =0.72). Com relação às MMP´s, não houve diferença estatística entre os dois grupos. Conclusões: Os níveis plasmáticos de NO encontram-se elevados em mulheres com DPC, especialmente naquelas com dor de origem visceral. Este fato pode ser considerado uma possibilidade no seguimento de pacientes com DPC, visto que pode ser usado como um marcador sérico para a doença. Já a dosagem das MMP-s não se mostrou útil como marcador plasmático para mulheres com DPC. / Background: Chronic pelvic pain is a disease of high prevalence and a complex pathophysiology. The diagnostic methods are often inadequate and, consequently, treatment and follow-up of women is quite difficult. Several diseases that present with chronic pain has an inflammatory profile, which has not yet been investigated for the topic. Aim: to quantify levels of nitric oxide (NO) and metalloproteinases 2 (MMP-2) and 9 (MMP-9) in plasma of women with chronic pelvic pain. Methods: 64 women were included in the sudy and divided into 02 groups: chronic pelvic pain and control group with 37 patients in the first group and 27 patients in the second group. Patients in the study group were followed at the Endoscopy and Pelvic Pain Unit and the control group patients were followed in the Contraception Unit of the Hospital of the Medical School of Ribeirão Preto University of São Paulo. We performed the measurement of clinical pain by a unidimensional scale (VAS) and a multidimensional scale (McGill). Anxiety and depression scales were also filled. Individuals with any evidence of inflammation, hypertension, smoking or use of hormonal contraceptives were excluded. Individuals taking medication for pain, such as painkillers or antiinflammatory drugs were asked to stop them 72 hours before entering the study. A blood sample was collected from 10 ml during the appointment. This material was stored in bottle itself with anti-coagulant (EDTA and / or heparin), processed immediately on site for plasma separation and stored in a -70 ºC freezer. The concentrations of species related to NO were measured in liquid, always in triplicate by the method of chemiluminescence, which is one of the most simple, sensitive and accurate available to measure NO. We used a NO analyzer (Sievers Model 280 NO Analyzer - Boulder, CO, USA), which allows the measurement of NO in quantities as small as 1 pmol. The activity of MMP-2 and MMP-9 in plasma was determined by the zymography method, which consists of an electrophoresis of the samples in an SDS / PAGE system, which includes the enzyme substrate (gelatin) in the gel separation, allowing the disclosure and quantification of enzyme activity. Results: Plasma NO levels were higher in CPP women than in controls (16.8 ± 7.9 versus 12.2 ± 2.4, respectively) (P=0.0016). Furthermore, plasma nitrate levels were higher in CPP women with evidence of pain of visceral origin than in CPP women with evidence of an exclusive somatic component or controls (19.2 ± 8.9 versus 12.4 ± 1.8 versus 12.2 ± 2.4, respectively) (P=0.0001). No correlation was detected between NO levels and duration of symptoms or intensity of pain. Regarding the MMP\'s, there was no statistical difference between the two groups. Conclusion: Plasma levels of NO are elevated in women with CPP, especially those with visceral pain. This fact can be considered an option in treating patients with CPP as it can be used as a serum marker for the disease. On the other hand, MMP´s did not turn out to be a good serum marker for women with CPP.

Background: Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder (FGID) which can be defined as chronic, relapsing visceral pain with bloating associated with change in bowel habit. It affects up to 10-15% of the adult population in the UK and is more common in females. The cost of IBS in terms of health care utilisation is substantial, exceeding £45.6 million per year in the UK alone, yet its pathophysiology is incompletely understood. Visceral pain is the main and most difficult symptom to manage in IBS and many IBS female sufferers compare it to labour pain in its severity. Modulating visceral pain in healthy volunteers and IBS patients is therefore an important research area. Non-invasive magnetic stimulation may play a crucial role in this respect. Aim:The aim of this study is to ascertain whether non-invasive repetitive magnetic stimulation applied to the motor cortex and/or lumbosacrum can modulate gastrointestinal pain originating from the anorectum. Methods:Participants: 16 healthy volunteers and 10 IBS patients aged 18 and above were included in the study.Questionnaires: Healthy volunteers and IBS patients were asked to complete anxiety and depression questionnaire and IBS patients were requested to fill in an IBS severity questionnaire.Motor measurements in healthy subjects: Single-pulse lumbosacral magnetic stimulation (LSMS) was applied to the lumbosacral area for the anal sphincter where the largest motor evoked potential (MEP) amplitude response was detected. Single-pulse transcranial magnetic stimulation (TMS) was then performed at the pre-determined resting motor thresholds (RMT) for the anal sphincter and the hand.Sensory measurements in healthy subjects and IBS patients: Electrical stimulation was used to assess the changes in sensory and pain thresholds in the anorectal area. The subjects were asked to score the pain intensity using five-point categorical rating scales. In addition they were asked to describe the pain experienced using a shortened form of the McGill Pain Questionnaire. Intervention: Healthy volunteers received 6 paradigms of magnetic stimulation in a randomised order i.e. 3 repetitive LSMSs (1 Hz, 10 Hz and sham) and 3 repetitive TMSs (1 Hz, 10 Hz and sham) to investigate their modulatory effects on visceral sensitivity and to determine which of these interventions is most effective. The most effective active interventions (1 Hz rLSMS and 10 Hz rTMS) together with one sham were then trialled in a randomised fashion on IBS patients.Post intervention: Motor excitabilities were repeated at 30 min after each intervention. The assessment of sensory and pain thresholds at anal sphincter and rectum were done immediately, 30 and 60 min after each intervention. Results:Application of 1 Hz rLSMS led to alterations of anal sphincter motor excitabilities and resulted in a significant increase in the amplitude of lumbosacal-anal motor evoked potentials (MEPs) in healthy volunteers recorded at 30 min post intervention. In healthy volunteers, 1 Hz rLSMS and 10 Hz rTMS caused a significant increase in the rectal pain thresholds experienced immediately, 30 and 60 min after each intervention. 10 Hz rLSMS and 1 Hz rTMS only led to a significant rise in rectal pain thresholds immediately after their application. Furthermore, there was a significant increase in the rectal pain thresholds immediately, 30 and 60 min following 1 Hz rLSMS and 10 Hz rTMS in IBS patients. Conclusion:The application of magnetic stimulation to the cortical and lumbosacral areas to modulate visceral pain is a new concept, which reduced rectal sensitivity to painful stimuli and offers a much needed new approach in the management of abdominal pain in patients with IBS.

No description available.

<p>Increased visceral pain is often seen in patients with gastrointestinal (GI) inflammation. Some studies, however, have suggested that such pain may persist after resolution of damage or inflammation. Despite the debilitating pain associated with GI inflammation, and its significant impact on affected individuals, few studies have addressed this issue. We hypothesized that altered visceral pain responses would persist after resolution of a bout of colitis in an animal model of colitis. We studied the pain responses to colorectal distention in Wistar rats with dinitrobenzene sulfonic acid (DNBS)-induced colitis, using changes in heart rate as an index of pain. Colonic inflammation had resolved by day 15 after DNBS administration. The assessment of colonic inflammation was based on histological scores, colonic tissue pro-inflammatory cytokine levels and myleoperoxidase activity. Rats examined at 15 days post-DNBS administration exhibited diminished pain responses to colorectal distention as compared to healthy rats. This was associated with significant increases in colonic tissue levels of IL-4 and IL-10 as compared to healthy rats, indicating a possible role for these anti-inflammatory cytokines in counteracting the generation of pain and hyperalgesia. We also studied the effects of hydrogen sulfide (H2S) in our animal model, by administering inhibitors of two of the key enzymes involved in the production of H2S. Our results demonstrated that inhibition of H2S production did not significantly alter the pain responses observed in rats at 15 days post-DNBS administration. In summary, our results demonstrate altered autonomic responses to colorectal distension following resolution of colitis. Further research on the role of anti-inflammatory cytokines and H2S may help to determine the mechanism underlying this effect.</p> / Master of Science (MSc)

La maladie de Crohn est caractérisée par l'alternance de phases de rémission et de poussées d'inflammation intestinale, de survenue imprévisible. L'objectif de la thèse était d'étudier l'effet, en terme d'activité cérébrale (IRMf), de l'incertitude dans l'anticipation de la survenue d'une douleur viscérale dans la MC en rémission. Les résultats majeurs montrent que dans la MC, cette phase est associée à une hyperactivation significative, par rapport au sujet sain, du cortex cingulaire antérieur et postérieur, de l'insula, du thalamus et de l'amygdale. Il s'agit de régions clés dans la gestion sensorielle, cognitive et émotionnelle de la douleur. En conclusion, la MC est associée à une hyperréactivité cérébrale liée au caractère incertain de la survenue d'une douleur viscérale. Ceci peut expliquer une plus grande vulnérabilité des patients face aux effets du stress sur leurs symptômes et le cours de leur maladie et l'efficacité des thérapies cognitivo-comportementales. / Crohn's disease (CD) is a chronic recurring inflammatory bowel disease, with unpredictable recurrence of flares; this unpredictability leads to great anticipatory anxiety and stress. The aim of this thesis was to study the effect, in terms of brain activity (fMRI), of uncertainty in the anticipation of visceral pain in quiescent CD. Le main results show that in CD, uncertainty is associated with hyperactivity in the anterior and posterior cingulate cortex, insula, thalamus and amygdala. These are key regions in the regulation of sensory, cognitive and emotional aspects of pain. In conclusion, CD comprises excessive cerebral reactivity linked to the uncertain character of the occurrence of visceral pain. This might account for the greater vulnerability of CD patients towards the effects of stress on their symptoms and the course of their disease, as well as the efficacy of behavioral therapies that aim at modifying the activity of limbic structures.