Analgésiques opiacés et les antidépresseurs tricycliques (ATC) sont deux grandes classes d'agents thérapeutiques qui sont utilisés pour soulager les symptômes dus au nociception chronique. Mais ces agents chimio thérapeutiques présentent des effets secondaires comme la constipation, dépendance physique plus soulagement de la douleur insuffisante. D'où des complications aggravent la nécessité de concevoir une molécule qui agit sur nouvelle cible en vue de surmonter les effets secondaires causés par la classe dit ci-dessus de médicaments. Pendant ce temps, la distribution du peptide lié au gène de la calcitonine (CGRP), un neuropeptide de 37 acides aminés deux dans le système nerveux central et périphérique et son rôle dans la douleur viscérale a ouvert une nouvelle porte d'entrée pour le traitement de la douleur nociceptive viscérale. Ainsi, l'objectif principal du projet de fusionner propriété antagoniste de CGRP ainsi que la propriété antidépresseur dans une seule molécule, pour fournir un effet de synergie et de traiter les maladies chroniques, inflammatoires et négligées comme le syndrome du côlon irritable (IBS) et la maladie de Crohn. / Opiate analgesics and tricyclic antidepressants (TCAs) are two major classes of therapeutic agents which are used to alleviate symptoms due to chronic nociception. But these chemotherapeutic agents pose side effects like constipation, physical dependence along with insufficient pain relief. Hence these complications aggravate the necessity of designing a molecule which acts on new target in order to overcome the side effects caused by the above said class of drugs. Meanwhile, the distribution of Calcitonin Gene Related Peptide (CGRP), a 37 amino acid neuropeptide both in central and peripheral nervous system and its role in visceral pain has opened up a new gateway for treating visceral nociceptive pain. Thus, the main aim of the project was to merge CGRP antagonist property as well as Antidepressant property in a single molecule, to provide a synergistic effect and to treat Chronic, inflammatory and neglected diseases like Irritable bowel syndrome (IBS) and Crohn’s disease.

No description available.

During orthodontic tooth movement, stress is applied on the dento-alveolar complex, initiating a biological response. This response results in the remodelling of the periodontal ligament and the alveolar bone. When a force is placed on a tooth, the periodontal ligament is stretched and compressed depending on the direction of force. On the side where the ligament is stretched, a response resulting in bone deposition is initiated. On the opposite side, where the periodontal ligament is compressed, a response resulting in the resorption of bone begins. These responses are believed to be modulated by factors that are derived from the immune or nervous systems. When stress is placed on the periodontal ligament, it is believed that nerve fibres and neuroreceptors within the tissue are distorted, leading to the release of neurotrophins and a common concomitant clinical response of pain and pressure. These neurotrophins may interact with cells within the dentoalveolar complex, including fibroblasts, endothelial and alveolar bone cells, resulting in the initiation of bone resorption via the activation of intracellular secondary messengers, which leads to cellular proliferation and differentiation. Neurotrophin levels may play a role in the modulation of cellular activity in the periodontal ligament during orthodontic movement. They are a family of protein polypeptides which are important in neural cell differentiation and survival. One relatively well studied member of the family, Nerve Growth Factor (NGF), is a polypeptide essential for supporting cholinergic innervation in the brain and sympathetic and sensory innervation in the peripheral tissues. Within the dento-alveolar complex, the function and localization of neurotrophins and their receptors are yet to be determined. Previous studies have shown that there is an increase in NGF expression in tissues in response to injury, suggesting that NGF expression may increase in regions within the dento-alveolar complex where inflammation and bone remodelling are occurring. A study showed elevated levels of NGF mRNA in human periodontal ligament cells in vitro during increased transcription and translation of the bone-related proteins alkaline phosphatase and osteopontin, suggesting NGF facilitates bone formation. The NGF may be modulating cellular activity within the periodontal ligament. O’Hara et al. used immunohistochemical staining in the rat dento-alveolar complex to show that there was evidence of an increase in NGF synthesis and release by certain cells or tissues within the region of alveolar bone remodelling during the initial injury response period to orthodontic tooth movement. Ho used a similar model to test the hypothesis that there may be a positive relationship between the presence of osteoclasts and pre-osteoclasts with areas of NGF localisation. He found no relationship between osteoclasts and areas of NGF; however, his findings showed areas of unknown tissue within the periodontal ligament that were associated with NGF. To date, the relationship between NGF and the cellular process of orthodontic tooth movement remains unknown. Further studies are required to describe the distribution of neurotrophins and neurotrophic receptors and cellular interactions within the rat dento-alveolar complex. / Thesis (D.Clin.Dent.) -- University of Adelaide, School of Dentistry, 2010

The distribution of nociceptive axons has not been fully determined in the whole rat and mouse heart. Previous anatomical studies have relied on sectioned tissue, which disrupts the organization of nerves and makes it more difficult to study their pattern, and most studies only used male tissues. We addressed this in the following two studies. First, flat-mounts of the right and left atria and ventricles, and the interventricular septum in rats were immunohistochemically labeled for calcitonin gene-related peptide (CGRP). Tissues were imaged with a high-quality microscope to generate complete montages and detailed images. We found that 1) CGRP-IR axons extensively innervated all regions of the atrial walls and the walls of the great vessels including the sinoatrial (SA) node region, auricles, atrioventricular (AV) node region, superior/inferior vena cava, left pre-caval vein, and pulmonary veins. 2) CGRP-IR axons formed varicose terminals around individual neurons in some cardiac ganglia. 3) Varicose CGRP-IR axons innervated the blood vessels. 4) CGRP-IR axons extensively innervated the right/left ventricular walls and interventricular septum. Second, flat-mount preparations of the left and right atria of male and female mice were labeled for CGRP, imaged, and digitally traced. The results show that 1) Large nerve bundle entry points and regional concentration of CGRP-IR axons were similar in both sexes. 2) The detailed distribution of nerves was digitized and mapped using sophisticated software and was similar between sexes. 3) Nerve density in the SA/AV node regions was not significantly different. 4) Morphometric parameters of varicosities in the auricle, SA/AV node regions was not significantly different between sexes. The distribution of CGRP-IR axons in the whole rat heart and the male/female mouse atria was shown for the first time at the single-cell/axon/varicosity resolution. Future studies will quantify the differences in CGRP-IR axon innervation between disease models, and other species.

No description available.

Interstitial cystitis is a chronic debilitating disease that causes pain and increased frequency of micturition, amongst other symptoms, without any identifiable cause. This disease affects a large number of the population, yet the etiology is still unknown. The present study aimed to characterize BDNF and CGRP—two neuropeptides that have both been proven to play an important role in the transmission of pain as well as in hypersensitivity. The signaling pathways regulating the expression of the two neuropeptides were also examined. Results revealed that BDNF protein expression levels increased in both L1 and L6 DRG following 48 hours post CYP-induced cystitis. CGRP protein expression levels decreased in L1 DRG, but increased in L6 DRG following 48 hours post CYP-induced cystitis. Examination of mRNA levels revealed an increase in the mRNA levels of both BDNF and CGRP in L6 DRG. NGF, a member of the neurotrophin family, mRNA levels also increased following 48 hour CYP-induced cystitis in the urinary bladder. Retrograde analysis revealed NGF possibly retrograde signaled to the DRG to increase BDNF and CGRP expression. Co-localization immunohistochemistry results revealed phospho-Akt co-localized with BDNF, but not with CGRP. Thus NGF retrograde signaling may activate the PI3-K/Akt cascade which may be involved in BDNF expression. CGRP expression may be via another signaling cascade.

Migraine affects ~15% of the US population (nearly 40 million people), making it one of the most common neurological disorders; however currently available therapeutic options for migraine relief are often ineffective. Moreover, acute and prophylactic drugs are both commonly associated with contraindications and serious side effects, and routine use of acute treatments may result in medication overuse-headaches. Elevated levels of the neuropeptide calcitonin gene-related peptide (CGRP) are known to be a primary factor in migraine pathogenesis, although the mechanisms by which CGRP expression becomes errantly modulated are unclear. CGRP is a product of the trigeminal ganglion and can be released both peripherally onto the dura mater, leading to neurogenic inflammation, and centrally at the spinal trigeminal nucleus, leading to neuromodulation. A great deal of CGRP-relevant migraine research has focused on the trigeminovascular system, but whether the cerebral cortex may have a role in migraine pathophysiology been less well studied. A subset of migraineurs experience a premonitory aura, which often manifests as a disturbance in one visual hemifield. An aberration called cortical spreading depression (CSD) is the likely electrophysiological substrate of the migraine aura, but whether CSD and CGRP are functionally related is not known. CSD is characterized by an initial transient wave of neuronal and glial depolarization, followed by a prolonged period of quiescence that is largely refractory to subsequent stimulation. Converging evidence supports a facilitatory role for cortical spreading depression (CSD) in migraine with and without aura, and CSD propagation has been shown to be dependent on functional CGRP receptors. Moreover, reported effects of CSD overlap with those of CGRP-mediated neurogenic inflammation. The experiments described herein seek to test the hypothesis that induction of CSD in vivo will lead to increased CGRP expression in the rodent cerebral cortex. Preliminary data in rats suggests that 3M KCl-induced CSD can trigger increased CGRP expression in the ipsilateral cortex. Preliminary data in mice has been less conclusive. Presented here are the data obtained from mice and rats, as well as speculation on the cause(s) of the differences in CGRP expression between species and how these findings relate to human studies.

The CGRP monoclonal antibodies are the first class of medication developed specifically for migraine prevention, in contrast to previous preventative medications, that were in the anti-hypertensive, anti-epileptic and anti-depressant class. There are two notable divisions within the CGRP inhibitor class: the CGRP monoclonal antibodies (CGRP mAbs), and the small molecule CGRP antagonists (gepants). This thesis conducts a retrospective analysis of notable clinical trials such as the ACHIEVE I, ACHIEVE II, LIBERTY, ARISE, STRIVE, PREEMPT, and COMPEL studies to determine the efficacy of CGRP inhibitors. In ACHIEVE I, 38.6% of participants in the 50 mg ubrogepant group experienced pain freedom 2 hours post dose (p=0.002) and in ACHIEVE II trial in the 50 mg ubrogepant group, 21.8% reported pain freedom 2 hours. In participants that received Rimegepant at a 75mg dose, 21%of participants reported more freedom from pain at 2 hours than placebo (p<0.0001).^40 In another study, participants received placebo, 50 mg and 100 mg of sumatriptan.^43 Results of the study showed that more than half of participants, 57%, in the 100 mg Sumatriptan group and exactly half of participants in the 50 mg group had pain relief at 2 hours post-dose.^43 In the LIBERTY trial, at 12 weeks, 30% of individuals that received erenumab reported a fifty percent or more reduction in the monthly number of migraine days than individuals in the placebo group (p=0.002).^45 In the STRIVE trial, the average number of migraine days experienced by the participant at baseline was 8.3, and was assessed by the 4th month through the 6th month. This baseline decreased to 5.1 days (a 3.2 difference) in the participants that received an injection of 70 mg of erenumab (p<0.001).^46 The participants that received an injection of 140 mg erenumab, decreased from the baseline to 4.6 days of migraine (a 3.7 difference) (p<0.001) . 46 Participants that received placebo reported the least change from baseline, only a 1.8 day change (p<0.001).46 In the ARISE Trial patients receiving erenumab experienced a change of 2.9 monthly migraine days, a 1.1 increase from the reported change of 1.8 days reported by study participants for the monthly migraine days in the placebo group (p<0.001).^47 The PREEMPT1 trial did not meet statistical significance for their primary endpoint or study measure, which was to assess for a mean change in monthly mean headache episode frequency between baseline and week 24 of the trial (p=0.344)^48. Participants in the PREEMPT2 trial experienced a reduction by 9 days when compared to placebo for the primary end point, frequency of headache days per 28 days relative to baseline (p<0.001)^49. In the COMPEL study, participants experienced -10.7 day reduction in headache days by 108 weeks (p<0.0001).^50 There are several advantages to CGRP mAbs. Patients are more likely to adhere to CGRP mAbs medication and tolerate this medication than other medication options^17, CGRP mAbs do not give rise to toxicity in the liver because these medications do not interact with the liver^17, and CGRP mAbs have a long duration in the human body as they have a half-life of 20 to 30 days which provides patients with the opportunity to not take the medication as frequently.^51 Another reason why CGRP mAbs are advantageous compared to traditional treatment options is that they have a strong affinity and specificity for the CGRP receptor or CGRP molecule. This high specificity prevents the medication from causing undesirable effects on other receptors^51.

Submitted by MARJORIE DE OLIVEIRA GALLINARI null (marjo_oliveira@hotmail.com) on 2016-05-20T00:08:43Z No. of bitstreams: 1 Versão final - Diessertação!.pdf: 4298608 bytes, checksum: d78e89bc5d747df80a571cc21dbf5c26 (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-05-23T19:19:29Z (GMT) No. of bitstreams: 1 gallinari_mo_me_arafo.pdf: 4298608 bytes, checksum: d78e89bc5d747df80a571cc21dbf5c26 (MD5) / Made available in DSpace on 2016-05-23T19:19:29Z (GMT). No. of bitstreams: 1 gallinari_mo_me_arafo.pdf: 4298608 bytes, checksum: d78e89bc5d747df80a571cc21dbf5c26 (MD5) Previous issue date: 2016-05-13 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo do presente estudo foi avaliar a influência da hidrocortison e do acetaminofeno na inflamação e na expressão de neuropeptídeos por meio da análise histopatológica e imunoistoquímica. Para tanto, 63 ratos foram divididos em 3 lotes de estudo (n=21) de acordo com a terapia de combate à dor: LI- controle, LII- administração tópica de hidrocortisona por 10 minutos, depois do tratamento clareador e LIII- administração via oral de acetaminofeno 30 minutos antes do tratamento clareador com peróxido de hidrogênio a 35% e depois de 12 em 12 horas. Em todos os grupos de estudo, na maxila esquerda foi realizado o tratamento clareador placebo e a maxila direita recebeu a 3 aplicações de 15 minutos de um gel clareador a base de peróxido de hidrogênio a 35%, totalizando 45 minutos de contato do gel clareador com substrato dentário. Os momentos de eutanásia dos animais foram imediatamente após, 24 e 48 horas após o tratamento clareador. Posteriormente à eutanásia dos animais, as peças foram processadas e o primeiro molar de cada maxila foi analisado histopatologicamente quanto ao grau de inflamação e por análise de imunoistoquímica para verificarmos a presença dos neuropeptídeos SP e CGRP. Os dados obtidos foram submetidos ao teste estatístico não paramétrico Kruskal Wallis seguido do teste de Dunn para comparações individuais, sendo observado na análise histopatológica total desorganização celular, extensas áreas de necrose nos grupos clareados, e o grupo que recebeu tratamento com otosporim apresentou melhores resultados. Na análise imuno-histoquimica, obteve imunomarcação positiva em todos os grupos, inclusive controle, porém nos grupos clareados a imunomarcação foi mais forte, sendo que o grupo que recebeu tratamento com otosporim apresentou os melhores resultados. Conclui-se que o uso da hidrocortisona após tratamento clareador minimiza os efeitos colaterais deste procedimento estético. / The aim of this study was to evaluate the influence of hydrocortisone and acetaminofen substances in inflammation and neuropeptide expression by histopathologic and immunohistochemical analysis. For this, 63 rats were divided into 3 batches of study (n = 21) according to combat pain therapy: Li control LII- topical administration of Otosporin® for 10 minutes after the bleaching treatment and administration route LIIIoral Tylenol® 30 minutes before the bleaching with hydrogen peroxide at 35% and then 12 for 12 hours. In all study groups in left maxilla was performed treatment whitener placebo and right jaw received three applications of 15 minutes a whitening gel 35% hydrogen peroxide base, totaling 45 minutes of contact of the whitening gel dental substrate. The times of the animals were euthanized immediately after 24 and 48 hours after the bleaching treatment. After the euthanasia of animals, the pieces were processed and the first molar of each jaw was analyzed histologically the degree of inflammation and analysis of immunohistochemistry to verify the presence of the neuropeptides SP and CGRP. The data were submitted to statistical nonparametric Kruskal Wallis test followed by Dunn's test for individual comparisons, being observed on histopathologic total cellular disorganization analysis, extensive areas of necrosis in whitened groups, and the group that received treatment with Otosporin® showed better results. In immunohistochemical analysis, obtained positive immunostaining in all groups, including control, but the whitened immunostaining groups was stronger, and the group that received treatment with Otosporin® showed the best results. We conclude that the use of Otosporin® after bleaching treatment minimizes the side effects of this cosmetic procedure. / FAPESP: 2015/01366-4

No momento em que há a agressão tecidual e a defesa inata é deflagrada, mediadores químicos são liberados no local afetado. Esses mediadores podem ser de origem celular tais como CGRP, VEGF e TGFß. Os objetivos desta pesquisa foram avaliar a expressão e distribuição de CGRP, TGFß e VEGF na gengiva do primeiro molar inferior esquerdo de rato no 7 e 14 dias após a indução por ligadura; a expressão e distribuição de CGRP, TGFß e VEGF na gengiva do dente contralateral correspondente sem ligadura, no 7 e 14 dias, e se a indução da periodontite por ligadura no dente experimental provoca uma inflamação na gengiva do dente contralateral correspondente no 7 e 14 dias após a ligadura. Para o desenvolvimento deste trabalho foram selecionados 15 ratos Rattus Novergicus, Albinus, Wistar. O grupo experimental de 12 ratos foi dividido em 2 subgrupos compostos por 6 ratos cada um deles distribuídos da seguinte maneira: os do subgrupo A1 permaneceram com a ligadura no primeiro molar inferior esquerdo por 7 dias e foram sacrificados; os do subgrupo A-2 -permaneceram com a ligadura no primeiro molar inferior esquerdo por 14 dias e foram sacrificados. Outros três animais constituíram o grupo controle. Após o sacrifício dos 12 animais dos grupos experimentais e controle suas mandíbulas foram colocadas em ácido etilenodiaminotetracético (EDTA) neutro para sofrerem descalcificação. Então foram processadas para inclusão em parafina e os cortes histológicos foram corados pela hematoxilina-eosina e submetidos à técnica imunohistoquímica para imunomarcação de CGRP, VEGF e TGFß. Aos 7 dias de ligadura observou-se na lâmina própria gengival, epitélio juncional e epitélio oral, expressiva marcação para CGRP. A expressão de VEGF foi intensa na lamina própria e com pouca ou nenhuma marcação no epitélio oral e juncional. O TGFß apresentou pouca marcação na lâmina própria ou nenhuma marcação no epitélio oral e juncional. Aos 14 dias de ligadura houve expressiva marcação de CGRP na lâmina própria, epitélios oral e juncional. O VEGF e o TGFß apresentaram muita marcação na lâmina própria e pouca ou nenhuma marcação no epitélio oral e juncional. Na gengiva dos dentes contralaterais nos 7 e 14 dias houve pouca marcação do CGRP do TGFß na lâmina própria e muita marcação do VEGF. Na gengiva dos dentes controle observou-se muita marcação do CGRP no epitélio juncional e oral e na lâmina própria. O TGFß e o VEGF se expressaram muito pouco ou não se expressaram. Devido à marcação expressiva do VEGF na lâmina própria dos dentes contralaterais, permanece inconclusiva a adequação do uso dos dentes contralaterais nos estudos experimentais das doenças periodontais, embora a expressão de TGFß e CGRP tenham sido menores nestes dentes. A maior marcação do CGRP, VEGF e TGFß nos animais com 14 dias de ligadura do que aos 7 dias demonstra a progressão do processo inflamatório crônico, não se observando processo de reparação cicatricial. / At the time of the tissue aggression the innate defense is triggered and the chemical mediators are released in the affected site. These mediators may have cell origin as the CGRP, VEGF and TGFß. The aim of this research were to evaluate the expression and distribution of the CGRP, VEGF and TGFß in the gingiva of the lower left first molar of rats in the seven and fourteen days after the ligature for inflammation induction; to analyse the expression and distribution of the CGRP, VEGF and TGFß in the gingiva of the correspondent counter lateral tooth without ligature in the seven and fourteen days and if the induction of periodontitis causes gingival inflammation of the counter lateral tooth after seven and fourteen days after ligature. For the development of this work were selected fifteen Rattus Novergicus, Albinus,Wistar. The experimental group of 12 rats was divided into 2 groups consisting of 6 rats each distributed of the following manner: the subgroup A1 remained with ligature in the first molar and left for 7 days before the sacrificed and the subgroup A-2 remained with the ligature for 14 days before the sacrificed. Another 3 animals constituted the control group. After the sacrificed of the 12 experimental and 3 control animals were immersedin neutral ethylenidiaminetetracetic (EDTA) for the decalcification. Then were processed for paraffin embedding. The sections were stained with hematoxylin-eosin and submitted to immunohistochemical techniques to imunostaining for CGRP, VEGF and TGFß. At 7 days of ligature was observed in the gingival lamina propria, junctional epithelium and oral epithelium, strong staining for CGRP. The intensive expression of VEGF occur in the lamina propria and scarce or no staining in the oral and junctional epithelium. The TGFß shows scarce staining in the lamina propria and no staining in the oral and junctional epithelium. At the fourteen days of ligature we observed strong staining of CGRP in the lamina propria and oral and junctional epithelium. The VEGF and TGFß show strong staining in the lamina propria and scarce or no staining in the junctional and oral epithelium. At the gingival of the counterlateral teeth in the 7 and 14 days there was scarce staining for CGRP and TGFß in the lamina propria and strong staining to VEGF. In the gingival of couterlateral teeth there was strong staining to CGRP in the junctional and oral epithelium and lamina propria. The TGFß and VEGF show scarce or none staining. Because the strong staining of VEGF in the lamina propria of the counterlateral teeth remain inconclusive the adequacy of the use of the counterlateral teeth in the experimental studies of the periodontal diseases, though the expression of the TGFß and CGRP has been smaller in this teeth. The bigger staining of CGRP, VEGF and TGFß in the animals with 14 days of ligature than those 7 days shows the evolution of the chronic inflammation process. We dont observe the process of tissue healing.