The potential anti-inflammatory and anti-nociceptive effects of rat hemopressin (PVNFKFLSH) in experimental arthritis.

Inflammatory arthritis, such as rheumatoid arthritis (RA), stands out as one of the main sources of pain and impairment to the quality of life. The use of hemopressin (PVNFKFLSH; Hp), an inverse agonist of type 1 cannabinoid receptor, has proven to be effective in producing analgesia in pain models, but its effect on neuro-inflammatory aspects of RA is limited. In this study, antigen-induced arthritis (AIA) was evoked by the intraarticular (i.art.) injection of methylated bovine serum albumin (mBSA) in male Sprague Dawley rats. Phosphate buffered saline (PBS)-injected ipsilateral knee joints or AIA contralateral were used as control. Nociceptive and inflammatory parameters such as knee joint oedema and leukocyte influx and histopathological changes were carried out in addition to the local measurement of interleukins (IL) IL-6, IL-1ß, tumor necrosis factor-α and the immunoreactivity of the neuropeptides substance P (SP) and calcitonin gene related peptide (CGRP) in the spinal cord (lumbar L3-5 segments) of AIA rats. For 4 days, AIA rats were treated daily with a single administration of saline, Hp injected (10 or 20 µg/day, i.art.), Hp given orally (20 µg/Kg, p.o.) or indomethacin (Indo; 5 mg/Kg, i.p.). In comparison to the PBS control group, the induction of AIA produced a significant and progressive mono-arthritis condition. The degree of AIA severity progressively compromised the normal walking pattern and impaired mobility over the next four days in relation to PBS-injected rats or contralateral knee joints. In AIA rats, the reduction of the distance between footprints and disturbances of gait evidenced signs of nociception. This response worsened at day 4, and a loss of footprint from the ipsilateral hind paw was evident. Daily treatment of the animals with Hp either i.art. (10 and 20 µg/knee) or p.o. (20 µg/Kg) as well as Indo (5 mg/Kg, i.p.) ameliorated the impaired mobility in a time-dependent manner (P < 0.05). In parallel, the AIA-injected ipsilateral knee joints reach a peak of swelling 24 h after AIA induction, which persisted over the next four days in relation to PBS-injected rats or contralateral knee joints. There was a significant but not dose-dependent inhibitory effect produced by all dosages and routes of Hp treatments on AIA-induced knee joint swelling (P < 0.05). In addition, the increased synovial levels of MPO activity, total leukocytes number and IL-6, but not IL-1ß, were significantly reduced by the lower i.art. dose of Hp. In conclusion, these results successfully demonstrate that Hp may represent a novel therapeutic strategy to treat RA, an effect which is unrelated to the proinflammatory actions of the neuropeptides CGRP and SP.

Potentiation of capsaicin-induced neurogenic inflammation by 5-HT7 receptors in the rat hind paw: Involvement of calcitonin gen-related peptide.

A decrease in the activation threshold of primary sensory neurons to transient receptor potential V1 (TRPV1) stimulation by serotonin 5-HT7 receptors has been reported but no confirmation if this might translate into facilitation of neurogenic inflammation has been provided. We analysed the modulation of capsaicin (CAP)-induced neurogenic inflammation in the rat hind paw by the selective 5-HT7 receptor agonist, LP-44, and the involvement of calcitonin gen-related peptide (CGRP) in this effect. Animals received intra-plantar injections (30 µL) of vehicle, CAP (0.05%, 0.1% and 0.2%), LP-44 (7.5 and 15 nmol) and the combination of LP-44 + CAP; then, the time course of the inflammatory responses was measured. The effect of the 5-HT7 receptor antagonist, SB-269970 (3 mg/kg, s.c.), on responses produced by LP-44 alone and combined with CAP was tested. As expected, CAP produced concentration- and time-dependent inflammatory responses in the hind paw. Interestingly, LP-44 by itself also produced inflammation in a concentration- and time-dependent manner, and magnified CAP-induced responses. Systemic pre-treatment with SB-269970 significantly blunted LP-44 (15 nmol)-induced inflammation as well as magnified inflammatory responses produced by the combination of LP-44 (7.5 and 15 nmol) + CAP (0.1%) thus confirming the involvement of 5-HT7 receptors. Finally, the non-peptide CGRP receptor antagonist, BIBN4096 (3 mg/kg, s.c.), strongly inhibited the potentiated inflammatory responses induced by LP-44 (7.5 and 15 nmol) + CAP (0.1%) thus substantiating their neurogenic nature. Thus, sensitization of CAP-sensitive primary sensory neurons by 5-HT7 receptors may result in facilitation of neurogenic inflammation involving CGRP in the rat hind paw.

Adrenomedullin and angiotensin II signaling pathways involved in the effects on cerebellar antioxidant enzymes activity.

Human adrenomedullin (AM) is a 52-amino acid peptide involved in cardiovascular control. AM has two specific receptors formed by the calcitonin-receptor-like receptor (CRLR) and receptor activity-modifying protein (RAMP) 2 or 3, known as AM1 and AM2 receptors, respectively. In addition, AM has appreciable affinity for the calcitonin gene-1 related peptide receptor (CGRP1), composed of CRLR/RAMP1. In brain, AM and their receptors are expressed in several localized areas, including the cerebellum. AM has been reported as an antioxidant. Little is known about the role of AM in the regulation of cerebellar reactive oxygen species (ROS) metabolism. We assessed the effect of AM on three antioxidant enzymes activity: catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) and on thiobarbituric acid reactive substances (TBARS) production in rat cerebellar vermis, as well the receptor subtypes involved in AM actions. Additionally, we evaluated the role of angiotensin II (ANG II), protein kinase A (PKA) activity, and protein kinase C/nicotinamide adenine dinucleotide phosphate oxidase (PKC/NAD(P)H) (oxidase) pathway. Sprague-Dawley rats were sacrificed by decapitation and cerebellar vermis was microdissected under stereomicroscopic control. CAT, GPx, SOD activity and TBARS production was determined spectrophotometrically. Our findings demonstrated that in cerebellar vermis, AM decreased and ANG II increased CAT, GPx and SOD activity and TBARS production. Likewise, AM antagonized ANG II-induced increase antioxidant enzyme activity. AM(22-50) and CGRP(8-37) blunted AM-induced decrease of antioxidant enzymes activity and TBARS production indicating that these actions are mediated through AM and CGRP1 receptors. Further, PKA inhibitor (PKAi) blunted AM action and apocynin and chelerythrine reverted ANG II action, suggesting that AM antioxidant action is mediated through stimulation of PKA activity, while ANG II-induced stimulation through PKC/NAD(P)H oxidase pathway. Our results support the role of AM in the regulation of cerebellar antioxidant enzymes activity and suggest a physiological role for AM in cerebellum.

Heteroreceptors Modulating CGRP Release at Neurovascular Junction: Potential Therapeutic Implications on Some Vascular-Related Diseases.

Calcitonin gene-related peptide (CGRP) is a 37-amino-acid neuropeptide belonging to the calcitonin gene peptide superfamily. CGRP is a potent vasodilator with potential therapeutic usefulness for treating vascular-related disease. This peptide is primarily located on C- and Aδ-fibers, which have extensive perivascular presence and a dual sensory-efferent function. Although CGRP has two major isoforms (α-CGRP and ß-CGRP), the α-CGRP is the isoform related to vascular actions. Release of CGRP from afferent perivascular nerve terminals has been shown to result in vasodilatation, an effect mediated by at least one receptor (the CGRP receptor). This receptor is an atypical G-protein coupled receptor (GPCR) composed of three functional proteins: (i) the calcitonin receptor-like receptor (CRLR; a seven-transmembrane protein), (ii) the activity-modifying protein type 1 (RAMP1), and (iii) a receptor component protein (RCP). Although under physiological conditions, CGRP seems not to play an important role in vascular tone regulation, this peptide has been strongly related as a key player in migraine and other vascular-related disorders (e.g., hypertension and preeclampsia). The present review aims at providing an overview on the role of sensory fibers and CGRP release on the modulation of vascular tone.

Influência do peróxido e do uso de diferentes substâncias de combate à dor na inflamação e na expressão neuropeptídeos pró-inflamatórios após o tratamento clareador / Effect of peroxide and the use of different substances fighting pain in inflammation and expression of neuropeptides pro-inflammatory after bleaching treatment

O objetivo do presente estudo foi avaliar a influência do Otosporin® e do Tylenol® 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 Otosporin® por 10 minutos, depois do tratamento clareador e LIII- administração via oral de Tylenol® 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 Otosporin® 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 Otosporin® apresentou os melhores resultados. Conclui-se que o uso do Otosporin® 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

Therapeutic antibodies against CGRP or its receptor.

CGRP is an extensively studied neuropeptide that has been implicated in the pathophysiology of migraine. While a number of small molecule antagonists against the CGRP receptor have demonstrated that targeting this pathway is a valid and effective way of treating migraine, off-target hepatoxicity and formulation issues have hampered the development for regulatory approval of any therapeutic in this class. The development of monoclonal antibodies to CGRP or its receptor as therapeutic agents has allowed this pathway to be re-investigated. Herein we review why CGRP is an ideal target for the prevention of migraine and describe four monoclonal antibodies against either CGRP or its receptor that are in clinical development for the treatment of both episodic and chronic migraine. We describe what has been publically disclosed about their clinical trials and future clinical development plans.

Pharmacological characterization of the relaxant effect induced by adrenomedullin in rat cavernosal smooth muscle

The aim of the present study was to determine the mechanisms underlying the relaxant effect of adrenomedullin (AM) in rat cavernosal smooth muscle (CSM) and the expression of AM system components in this tissue. Functional assays using standard muscle bath procedures were performed in CSM isolated from male Wistar rats. Protein and mRNA levels of pre-pro-AM, calcitonin receptor-like receptor (CRLR), and Subtypes 1, 2 and 3 of the receptor activity-modifying protein (RAMP) family were assessed by Western immunoblotting and quantitative real-time polymerase chain reaction, respectively. Nitrate and 6-keto-prostaglandin F1α (6-keto-PGF1α; a stable product of prostacyclin) levels were determined using commercially available kits. Protein and mRNA of AM, CRLR, and RAMP 1, -2, and -3 were detected in rat CSM. Immunohistochemical assays demonstrated that AM and CRLR were expressed in rat CSM. AM relaxed CSM strips in a concentration-dependent manner. AM22-52, a selective antagonist for AM receptors, reduced the relaxation induced by AM. Conversely, CGRP8-37, a selective antagonist for calcitonin gene-related peptide receptors, did not affect AM-induced relaxation. Preincubation of CSM strips with NG-nitro-L-arginine-methyl-ester (L-NAME, nitric oxide synthase inhibitor), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, quanylyl cyclase inhibitor), Rp-8-Br-PET-cGMPS (cGMP-dependent protein kinase inhibitor), SC560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethyl pyrazole, selective cyclooxygenase-1 inhibitor], and 4-aminopyridine (voltage-dependent K+ channel blocker) reduced AM-induced relaxation. On the other hand, 7-nitroindazole (selective neuronal nitric oxide synthase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), H89 (protein kinase A inhibitor), SQ22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, adenylate cyclase inhibitor], glibenclamide (selective blocker of ATP-sensitive K+ channels), and apamin (Ca2+-activated channel blocker) did not affect AM-induced relaxation. AM increased nitrate levels and 6-keto-PGF1α in rat CSM. The major new contribution of this research is that it demonstrated expression of AM and its receptor in rat CSM. Moreover, we provided evidence that AM-induced relaxation in this tissue is mediated by AM receptors by a mechanism that involves the nitric oxide-cGMP pathway, a vasodilator prostanoid, and the opening of voltage-dependent K+ channels.

Pharmacological characterization of the relaxant effect induced by adrenomedullin in rat cavernosal smooth muscle.

The aim of the present study was to determine the mechanisms underlying the relaxant effect of adrenomedullin (AM) in rat cavernosal smooth muscle (CSM) and the expression of AM system components in this tissue. Functional assays using standard muscle bath procedures were performed in CSM isolated from male Wistar rats. Protein and mRNA levels of pre-pro-AM, calcitonin receptor-like receptor (CRLR), and Subtypes 1, 2 and 3 of the receptor activity-modifying protein (RAMP) family were assessed by Western immunoblotting and quantitative real-time polymerase chain reaction, respectively. Nitrate and 6-keto-prostaglandin F(1α) (6-keto-PGF(1α); a stable product of prostacyclin) levels were determined using commercially available kits. Protein and mRNA of AM, CRLR, and RAMP 1, -2, and -3 were detected in rat CSM. Immunohistochemical assays demonstrated that AM and CRLR were expressed in rat CSM. AM relaxed CSM strips in a concentration-dependent manner. AM(22-52), a selective antagonist for AM receptors, reduced the relaxation induced by AM. Conversely, CGRP(8-37), a selective antagonist for calcitonin gene-related peptide receptors, did not affect AM-induced relaxation. Preincubation of CSM strips with N(G)-nitro-L-arginine-methyl-ester (L-NAME, nitric oxide synthase inhibitor), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, quanylyl cyclase inhibitor), Rp-8-Br-PET-cGMPS (cGMP-dependent protein kinase inhibitor), SC560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethyl pyrazole, selective cyclooxygenase-1 inhibitor], and 4-aminopyridine (voltage-dependent K(+) channel blocker) reduced AM-induced relaxation. On the other hand, 7-nitroindazole (selective neuronal nitric oxide synthase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), H89 (protein kinase A inhibitor), SQ22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, adenylate cyclase inhibitor], glibenclamide (selective blocker of ATP-sensitive K(+) channels), and apamin (Ca(2+)-activated channel blocker) did not affect AM-induced relaxation. AM increased nitrate levels and 6-keto-PGF1α in rat CSM. The major new contribution of this research is that it demonstrated expression of AM and its receptor in rat CSM. Moreover, we provided evidence that AM-induced relaxation in this tissue is mediated by AM receptors by a mechanism that involves the nitric oxide-cGMP pathway, a vasodilator prostanoid, and the opening of voltage-dependent K(+) channels.

Cardiovascular effects of the intracerebroventricular injection of adrenomedullin: roles of the peripheral vasopressin and central cholinergic systems

Our objective was to investigate in conscious Sprague-Dawley (6-8 weeks, 250-300 g) female rats (N = 7 in each group) the effects of intracerebroventricularly (icv) injected adrenomedullin (ADM) on blood pressure and heart rate (HR), and to determine if ADM and calcitonin gene-related peptide (CGRP) receptors, peripheral V1 receptors or the central cholinergic system play roles in these cardiovascular effects. Blood pressure and HR were observed before and for 30 min following drug injections. The following results were obtained: 1) icv ADM (750 ng/10 µL) caused an increase in both blood pressure and HR (DMAP = 11.8 ± 2.3 mmHg and ΔHR = 39.7 ± 4.8 bpm). 2) Pretreatment with a CGRP receptor antagonist (CGRP8-37) and ADM receptor antagonist (ADM22-52) blocked the effect of central ADM on blood pressure and HR. 3) The nicotinic receptor antagonist mecamylamine (25 µg/10 µL, icv) and the muscarinic receptor antagonist atropine (5 µg/10 µL, icv) prevented the stimulating effect of ADM on blood pressure. The effect of ADM on HR was blocked only by atropine (5 µg/10 µL, icv). 4) The V1 receptor antagonist [β-mercapto-β-β-cyclopentamethylenepropionyl¹, O-me-Tyr²,Arg8]-vasopressin (V2255; 10 µg/kg), that was applied intravenously, prevented the effect of ADM on blood pressure and HR. This is the first study reporting the role of specific ADM and CGRP receptors, especially the role of nicotinic and muscarinic central cholinergic receptors and the role of peripheral V1 receptors in the increasing effects of icv ADM on blood pressure and HR.

Cardiovascular effects of the intracerebroventricular injection of adrenomedullin: roles of the peripheral vasopressin and central cholinergic systems.

Our objective was to investigate in conscious Sprague-Dawley (6-8 weeks, 250-300 g) female rats (N = 7 in each group) the effects of intracerebroventricularly (icv) injected adrenomedullin (ADM) on blood pressure and heart rate (HR), and to determine if ADM and calcitonin gene-related peptide (CGRP) receptors, peripheral V1 receptors or the central cholinergic system play roles in these cardiovascular effects. Blood pressure and HR were observed before and for 30 min following drug injections. The following results were obtained: 1) icv ADM (750 ng/10 µL) caused an increase in both blood pressure and HR (DMAP = 11.8 ± 2.3 mmHg and ΔHR = 39.7 ± 4.8 bpm). 2) Pretreatment with a CGRP receptor antagonist (CGRP8-37) and ADM receptor antagonist (ADM22-52) blocked the effect of central ADM on blood pressure and HR. 3) The nicotinic receptor antagonist mecamylamine (25 µg/10 µL, icv) and the muscarinic receptor antagonist atropine (5 µg/10 µL, icv) prevented the stimulating effect of ADM on blood pressure. The effect of ADM on HR was blocked only by atropine (5 µg/10 µL, icv). 4) The V1 receptor antagonist [ß-mercapto-ß-ß-cyclopentamethylenepropionyl¹, O-me-Tyr²,Arg8]-vasopressin (V2255; 10 µg/kg), that was applied intravenously, prevented the effect of ADM on blood pressure and HR. This is the first study reporting the role of specific ADM and CGRP receptors, especially the role of nicotinic and muscarinic central cholinergic receptors and the role of peripheral V1 receptors in the increasing effects of icv ADM on blood pressure and HR.

Relações entre eficiência alimentar, características de carcaça e qualidade de carne de novilhos Nelore confinados / Feed efficiency and its relationships with profitability, carcass characteristics and meat quality of Nellore steers

Embora possua o maior rebanho comercial do mundo, o Brasil está longe de liderar pesquisas relacionadas ao melhoramento genético de bovinos de corte para eficiência alimentar. Alguns países investiram em estudos para seleção de animais eficientes e identificação de marcadores moleculares associados a esta característica. A seleção genética de animais eficientes representaria economia de alimentos, reduziria o impacto ambiental e aumentaria a rentabilidade da atividade. Obviamente é um desafio identificar animais mais eficientes em um rebanho de 205 milhões de cabeças, pois, além da mensuração ser cara, os resultados são obtidos a longo prazo. As associações entre eficiência alimentar e desempenho de bovinos de corte são conhecidas, porém avaliações envolvendo os impactos da seleção para eficiência sobre composição corporal e qualidade de carne de novilhos Nelore ainda são incipientes. O objetivo neste trabalho foi estudar relações entre eficiência alimentar, composição corporal, características de carcaça e qualidade da carne em bovinos Nelore confinados. Foram confinados, em dois anos, 322 novilhos para avaliação de características de carcaça e qualidade de carne. Deste total, 159 foram confinados em baias individuais, dos quais 92 foram avaliados quanto ao consumo alimentar residual (CAR). Foram calculadas a eficiência alimentar, eficiência parcial de crescimento, taxa relativa de crescimento, índice de Kleiber e CAR. No abate foram obtidos dados de rendimento, comprimento e profundidade de carcaça, área de olho de lombo, espessura de gordura subcutânea, peso do coração, fígado, rins e gordura interna. Foram obtidas variáveis de qualidade da carne de amostras não maturadas do Longissimus dorsi (pH, força de cisalhamento, cor da carne e gordura, perdas por cocção, capacidade de retenção de água e extrato etéreo). Foram analisados os coeficientes de regressão linear entre as variáveis avaliadas e o CAR. Animais mais eficientes apresentaram consumo no mínimo 12% menor que os ineficientes (P 0,05). No entanto, a diminuição do CAR reduziu a deposição de gordura subcutânea, avaliada por ultrassom (P<0,05). Consistente com essa redução, o conteúdo de gordura intramuscular diminuiu (P = 0,08), indicando uma relação positiva entre CAR e deposição de gordura. A energia retida, estimada pela composição corporal, foi reduzida em 0,4 Mcal / dia. Isto significa que mudanças na composição corporal foram capazes de explicar 36% das diferenças no consumo de alimentos entre animais eficientes e ineficientes quanto ao CAR. A margem de contribuição (lucro) foi positivamente relacionada ao CAR (P<0,05), mas foi muito melho...

Although Brazil has the largest commercial beef cattle herd of the world, research on genetic improvement of beef cattle for feed efficiency is incipient. Some countries have invested in studies to identify feed efficient animals and to find molecular markers associated with this characteristic. Genetic selection of efficient animals can resulting lower costs, reduced environmental impact and increased profitability. Obviously identifying the most efficient animals is a challenge in a herd of 205 million head as measurement of feed intake is expensive. Associations between feed efficiency and performance of beef cattle are known, but possible impacts of selection for efficiency on body composition and meat quality of Nellore are still unknown. The objective of this study was to investigate relationships of feed efficiency with corporal composition, carcass characteristics and meat quality in Nellore steers finished in the feedlot. To evaluate carcass characteristics and meat quality, 322 steers were fed over a two year period. Of this total, 159 were housed in individual pens and 92 were evaluated for feed consumption. Feed efficiency, residual feed intake (RFI) partial efficiency of growth, relative growth rate and Kleiber´s index were calculated. Variables evaluated were: dressing, yield carcass linear measurements, loin eye area, fat thickness and weights of heart, liver, kidneys and internal fat. Longissimus dorsi pH, shear force, color of meat and fat, cooking losses, holding water capacity and ether extract were collected. The linear regression coefficients between these variables and RFI were calculated. Efficient and inefficient animals were separated as those that had 1 unit difference in RFI. The efficient animals intake were 12% lower than the inefficient animals (P0.05). Consistent with the reduction in subcutaneous fat deposition there was a decrease in the fat content (P=0.08), indicating a positive relationship between RFI and fat deposition. The retained energy, estimated by the difference in body composition, was reduced by 0.4 Mcal/day in inefficient vs efficient animals. This means that the change in estimated body composition was able to explain 36% of the difference in feed intake between RFI efficient and RFI inefficient animals. Individual profit was calculated and it was positively associated to RFI (P<0.05). However, profit had much greater association with feed conversion efficiency (r=0.75) as well as with weight gain (r=0.53). The association of RFI with profitability was low (0.23) and about as good as the measurement of loin eye area (r=0.20). In this work RFI ...

The role of CGRP in the pathophysiology of migraine and efficacy of CGRP receptor antagonists as acute antimigraine drugs.

Migraine is a highly prevalent neurovascular disorder that can be provoked by infusion of calcitonin gene-related peptide (CGRP). CGRP, a neuropeptide released from activated trigeminal sensory nerves, dilates intracranial and extracranial blood vessels and centrally modulates vascular nociception. On this basis, it has been proposed that: (i) CGRP may play an important role in the pathophysiology of migraine; and (ii) blockade of CGRP receptors may abort migraine. With the advent of potent and selective CGRP receptor antagonists, the importance of CGRP in the pathophysiology of migraine and the therapeutic principle of CGRP receptor antagonism were clearly established. Indeed, both olcegepant (BIBN4096BS, given intravenously) and telcagepant (MK-0974, given orally) have been shown to be safe, well tolerated and effective acute antimigraine agents in phase I, phase II, and for telcagepant phase III, studies. However, recent data reported elevated liver transaminases when telcagepant was dosed twice daily for three months for the prevention of migraine rather than acutely. The potential for a specific acute antimigraine drug, without producing vasoconstriction or vascular side effects and with an efficacy comparable to triptans, is enormous. The present review will discuss the role of CGRP in the pathophysiology of migraine and the various treatment modalities that are currently available to target this neuropeptide.

Calcitonin gene-related peptide receptor expression in alternatively activated monocytes/macrophages during irreversible pulpitis.

The purpose of this study was to quantify the percentage and the mean fluorescence intensity of viable alternatively activated monocytes/macrophages (AAMø) CD163+ positive for calcitonin gene-related peptide receptor (CGRPr) within the total AAMø population in human dental pulp. Pulp tissue samples were collected from teeth with a clinical diagnosis of irreversible pulpitis (n = 13), pulps with induced inflammation (n = 13), and normal pulps (n = 13). All samples were labeled to identify positive cells for CGRPr and CD163 using a flow cytometry assay. Results demonstrated that a high percentage of total viable AAMø CD163+ expressed CGRPr on their membranes (72.12% in healthy pulp, 62.20% in irreversible pulpitis, and 58.01% in induced pulpitis). Significant differences were found between mean AAMø CD163+ fluorescence for CGRPr according to pulp condition, being greater in irreversible pulpitis. It can be concluded that AAMø CD163+ are expressed during normal and inflammatory processes, supporting the hypothesis that they could exercise an anti-inflammatory action that could be controlled by CGRP signaling after its binding.

Evidence for the role of neurogenic inflammation components in trypsin-elicited scratching behaviour in mice.

BACKGROUND AND PURPOSE: We investigated the mechanisms underlying the pruritogenic response induced by trypsin in mice, to assess the relevance of neurogenic inflammation components in this response. EXPERIMENTAL APPROACH: Itching was induced by an intradermal injection of trypsin in the mouse neck. The animals were observed for 40 min and their scratching behaviour was quantified. KEY RESULTS: Trypsin-induced itching was blocked by the lima bean trypsin inhibitor, the selective proteinase-activated receptor-2 (PAR-2) antagonist FSLLRY and PAR-2 receptor desensitization. An important involvement of mast cells was observed, as chronic pretreatment with the mast cell degranulator compound 48/80 or the mast cell stabilizer disodium cromoglycate prevented scratching. Also, trypsin response was inhibited by the selective COX-2 inhibitor celecoxib and by the selective kinin B2 (FR173657) and B1 (SSR240612) receptor antagonists. Moreover, an essential role for the mediators of neurogenic inflammation was established, as the selective NK1 (FK888), NK3 (SR142801) and calcitonin gene-related peptide (CGRP(8-37) fragment) receptor antagonists inhibited trypsin-induced itching. Similarly, blockade of transient receptor potential vanilloid 1 (TRPV1) receptors by the selective TRPV1 receptor antagonist SB366791, or by genetic deletion of TRPV1 receptor reduced this behaviour in mice. C-fibre desensitization showed a very similar result. CONCLUSIONS AND IMPLICATIONS: Trypsin intradermal injection proved to be a reproducible model for the study of itching and the involvement of PAR-2 receptors. Also, trypsin-induced itching seems to be widely dependent on neurogenic inflammation, with a role for TRPV1 receptors. In addition, several other mediators located in the sensory nerves and skin also seem to contribute to this process.

Papel de la adrenomedulina cerebelosa en la hipertensión arterial / Paper of the adrenomedulina cerebelosa in the arterial hypertension

La adrenomedulina (AM) y el péptido relacionado con el gen de la calcitonina (CGRP) pertenecen a la superfamilia de los péptidos de CGRP. En el SNC, los sitios de unión para la AM y el CGRP se encuentran presentes en áreas hipotalámicas y en la corteza cerebelosa de la rata. La administración central de AM o de CGRP en ratas induce diuresis, natriuresis e incremento de la presión arterial. El papel de la AM en el cerebelo se desconoce. Con el fin de establecer la posible relación de la AM y CGRP cerebelosa y la regulación cardiovascular, en el presente estudio evaluamos la densidad de sitios de unión para la AM y el CGRP en el cerebelo de ratas espontáneamente hipertensas (SHR) y sus controles normotensos Wistar Kyoto (WKY) adultos de 16 semanas, mediante el uso de técnicas autoradiografícas y empleando 125I-hCGRPα y 125I-hAM13-52 como radioligandos. Los cortes coronales de cerebelo fueron incubados con 35 pM de [125I]-hCGRPα o [125I]-hAM13-52, durante 90 y 120 minutos, respectivamente. La unión no específica fue determinada en presencia de 1µM del ligando no marcado. El análisis densitométrico demostró que existe una colocalización de los sitios de unión para el [125I]-hCGRPα y la [125I]-hAM13-52 en la corteza cerebelosa. En el cerebelo la unión de la [125I]-hAM13-52 en las ratas SHR fue significativamente mayor que las WKY, indicando una mayor expresión de los receptores para la AM en el cerebelo de animales hipertensos. En relación a la unión de [125I]-hCGRPα, se observó también un pequeño incremento significativo en las ratas SHR en relación a las WKY. Con el fin de establecer la posible vía de señalización de la AM en la corteza cerebelosa, se evaluó la actividad de la óxido nítrico sintasa inducida por la AM.

Role of CGRP and GABA in the hypotensive effect of intrathecally administered anandamide to anesthetized rats.

In urethane-anesthetized rats the intrathecal (i.t.) injection of 100 nmol anandamide produced a hypotensive effect (-19.3+/-1.6 mm Hg; n=6) that was mimicked by i.t. administration of 0.25 nmol calcitonin gene-related peptide (CGRP; -26.2+/-1.8 mm Hg, n=4). Both effects were antagonized either by the CGRP receptor antagonist CGRP(8-37) (5 nmol; i.t.) or by the gamma-aminobutyric acid (GABA)(A) receptor antagonist bicuculline (8.8 nmol, i.t) or by the GABA(B) receptor antagonist 2-hydroxy saclofen (110 nmol; i.t.). On the contrary, blockade of spinal CGRP receptors by CGRP(8-37) did not modify the hypotensive response to either the GABA(A)-receptor agonist muscimol (8.8 nmol; i.t.) or the GABA(B)-receptor agonist baclofen (100 nmol; i.t). This result suggests a unidirectional effect of CGRP on the GABAergic system. The response to anandamide remained unaltered after acute inhibition of nitric oxide (NO) synthase activity by either i.t. (1 micromol) or i.v. (10 mg/kg) injection of N(G)-nitro-L-arginine methyl ester (L-NAME), but increased significantly after long-term L-NAME administration (70 mg/kg/day; four weeks; p.o.), thus suggesting compensatory changes in cardiovascular homeostasis. It is proposed that the hypotensive effect of anandamide in urethane-anesthetized rats could involve the release of CGRP followed by the release of GABA in the spinal cord. NO does not appear to have a direct participation in the spinal mechanisms involved in the decrease of the blood pressure caused by anandamide.

Calcitonin gene-related peptide inhibits local acute inflammation and protects mice against lethal endotoxemia.

Calcitonin gene-related peptide (CGRP), a potent vasodilatory peptide present in central and peripheral neurons, is released at inflammatory sites and inhibits several macrophage, dendritic cell, and lymphocyte functions. In the present study, we investigated the role of CGRP in models of local and systemic acute inflammation and on macrophage activation induced by lipopolysaccharide (LPS). Intraperitoneal pretreatment with synthetic CGRP reduces in approximately 50% the number of neutrophils in the blood and into the peritoneal cavity 4 h after LPS injection. CGRP failed to inhibit neutrophil recruitment induced by the direct chemoattractant platelet-activating factor, whereas it significantly inhibited LPS-induced KC generation, suggesting that the effect of CGRP on neutrophil recruitment is indirect, acting on chemokine production by resident cells. Pretreatment of mice with 1 mug of CGRP protects against a lethal dose of LPS. The CGRP-induced protection is receptor mediated because it is completely reverted by the CGRP receptor antagonist, CGRP 8-37. The protective effect of CGRP correlates with an inhibition of TNF-alpha and an induction of IL-6 and IL-10 in mice sera 90 min after LPS challenge. Finally, CGRP significantly inhibits LPS-induced TNF-alpha released from mouse peritoneal macrophages. These results suggest that activation of the CGRP receptor on macrophages during acute inflammation could be part of the negative feedback mechanism controlling the extension of acute inflammatory responses.

Calcitonin gene-related peptide receptor expression in healthy and inflamed human pulp tissue.

AIM: To use radioreceptor analysis for comparing calcitonin gene-related peptide (CGRP) receptor expression in human pulp tissue samples collected from teeth having a clinical diagnosis of acute irreversible pulpitis, healthy pulps and teeth with induced inflammation. METHODOLOGY: Six pulp samples were obtained from teeth having a clinical diagnosis of acute irreversible pulpitis. Another eight pulp samples were obtained from healthy premolars where extraction was indicated for orthodontic purposes. In four of these premolars, inflammation was induced prior to pulp collection. All the samples were processed and labelled with 125I-CGRP. Binding sites were identified by 125I-CGRP and standard CGRP competition assays. RESULTS: CGRP receptor expression was found in all human pulp tissue samples. Most receptors were found in the group of pulps from teeth having a clinical diagnosis of acute irreversible pulpitis, followed by the group of pulps having induced inflammation. The least number of receptors was expressed in the group of healthy pulps. The Kruskal-Wallis and Mann-Whitney (post-hoc) tests showed statistically significant differences between the groups (P < 0.05). CONCLUSION: CGRP receptor expression in human pulp tissue is significantly increased during inflammatory phenomena such as acute irreversible pulpitis.

Peptídeo relacionado ao gene da calcitonina por iontoforese na viabilidade de retalho cutâneo randômico em ratos / Calcitonin gene-related peptide by iontophoresis on the viability of the randon skin flaps in rats

Objetivo: Investigar o efeito da administração tópica do peptídeo relacionado ao gene da calcitonina (CGRP) por iontoforese na viabilidade de retalho cutâneo randômico em ratos. Métodos: Sessenta ratos Wistar EPM-1, adultos e machos foram submetidos a retalho cutâneo randômico. Os animais foram distribuídos aleatoriamente em quatro grupos. Nos animais do grupo 1 (controle, n=15) realizou-se simulação de estímulo elétrico, no grupo 2 (iontoforese placebo, n=15) os animais foram submetidos à corrente contínua, no grupo 3 (controle de absorção, n=15) os animais receberam simulação de estímulo elétrico com CGRP e, por fim os animais do grupo 4 (tratado, n=15) foram tratados com iontoforese de CGRP. Em todos os grupos estes procedimentos foram realizados imediatamente após a técnica operatória e nos dois dias subsequentes. A porcentagem da área de necrose foi avaliada no sétimo dia de pós-operatório. Resultados: A média das porcentagens das áreas de necrose foram: grupo 1- 48 por cento, grupo 2 - 51 por cento, grupo 3 - 46 por cento e, grupo 4 - 28 por cento. A análise estatística, através do teste de Kruskal-Wallis, evidenciou diferença significante (p<0,001). Conclusão: a administração tópica de CGRP por iontoforese é eficaz em aumentar a viabilidade de retalho cutâneo randômico em ratos.

Involvement of spinal neurokinins, excitatory amino acids, proinflammatory cytokines, nitric oxide and prostanoids in pain facilitation induced by Phoneutria nigriventer spider venom.

The major local symptom of Phoneutria nigriventer envenomation is an intense pain, which can be controlled by infiltration with local anesthetics or by systemic treatment with opioid analgesics. Previous work showed that intraplantar (i.pl) injection of Phoneutria nigriventer venom in rats induces hyperalgesia, mediated peripherally by tachykinin and glutamate receptors. The present study examined the spinal mechanisms involved in pain-enhancing effect of this venom. Intraplantar injection of venom into rat hind paw induced hyperalgesia. This phenomenon was inhibited by intrathecal (i.t.) injection of tachykinin NK1 (GR 82334) or NK2 (GR 94800) receptor antagonists, a calcitonin gene-related peptide (CGRP) receptor antagonist (CGRP8-37) and N-methyl-D-aspartate (NMDA; MK 801 and AP-5), non-NMDA ionotropic (CNQX), or metabotropic (AIDA and MPEP) glutamate receptor antagonists, suggesting the involvement of spinal neurokinins and excitatory amino acids. The role of proinflammatory cytokines, nitric oxide (NO), and prostanoids in spinally mediated pain facilitation was also investigated. Pharmacological blockade of tumour necrosis factor-alpha (TNFalpha) or interleukin-1beta (IL-1beta) reduced the hyperalgesic response to venom. Intrathecal injection of L-N6-(1-iminoethyl)lysine (L-NIL), but not of 7-nitroindazole (7-NI), inhibited hyperalgesia induced by the venom, indicating that NO, generated by the activity of the inducible form of nitric oxide synthase, also mediates this phenomenon. Furthermore, indomethacin, an inhibitor of cyclooxigenases (COX), or celecoxib, a selective inhibitor of COX-2, abolished venom-induced hyperalgesia, suggesting the involvement of spinal prostanoids in this effect. These data indicate that the spinal mechanisms of pain facilitation induced by Phoneutria nigriventer venom involves a plethora of mediators that may cooperate in the genesis of venom-induced central sensitization.