Biological and Molecular Docking Evaluation of a Benzylisothiocyanate Semisynthetic Derivative From Moringa oleifera in a Pre-clinical Study of Temporomandibular Joint Pain.

Objective: Moringa oleifera possesses multiple biological effects and the 4-[(4'-O-acetyl-α-L- rhamnosyloxy) benzyl] isothiocyanate accounts for them. Based on the original isothiocyanate molecule we obtained a semisynthetic derivative, named 4-[(2',3',4'-O-triacetyl-α-L-rhamnosyloxy) N-benzyl] hydrazine carbothioamide (MC-H) which was safe and effective in a temporomandibular joint (TMJ) inflammatory hypernociception in rats. Therefore, considering that there is still a gap in the knowledge concerning the mechanisms of action through which the MC-H effects are mediated, this study aimed to investigate the involvement of the adhesion molecules (ICAM-1, CD55), the pathways heme oxygenase-1 (HO-1) and NO/cGMP/PKG/K+ ATP, and the central opioid receptors in the efficacy of the MC-H in a pre-clinical study of TMJ pain. Methods: Molecular docking studies were performed to test the binding performance of MC-H against the ten targets of interest (ICAM-1, CD55, HO-1, iNOS, soluble cGMP, cGMP-dependent protein kinase (PKG), K+ ATP channel, mu (µ), kappa (κ), and delta (δ) opioid receptors). In in vivo studies, male Wistar rats were treated with MC-H 1 µg/kg before TMJ formalin injection and nociception was evaluated. Periarticular tissues were removed to assess ICAM-1 and CD55 protein levels by Western blotting. To investigate the role of HO-1 and NO/cGMP/PKG/K+ ATP pathways, the inhibitors ZnPP-IX, aminoguanidine, ODQ, KT5823, or glibenclamide were used. To study the involvement of opioid receptors, rats were pre-treated (15 min) with an intrathecal injection of non-selective inhibitor naloxone or with CTOP, naltrindole, or norbinaltorphimine. Results: All interactions presented acceptable binding energy values (below -6.0 kcal/mol) which suggest MC-H might strongly bind to its molecular targets. MC-H reduced the protein levels of ICAM-1 and CD55 in periarticular tissues. ZnPP-IX, naloxone, CTOP, and naltrindole reversed the antinociceptive effect of MC-H. Conclusion: MC-H demonstrated antinociceptive and anti-inflammatory effects peripherally by the activation of the HO-1 pathway, as well as through inhibition of the protein levels of adhesion molecules, and centrally by µ and δ opioid receptors.

Sulfated polysaccharide from the green marine algae Caulerpa racemosa reduces experimental pain in the rat temporomandibular joint.

Temporomandibular disorder is a clinical painful condition in the temporomandibular joint (TMJ) region. The purified sulfated polysaccharide from the green marine algae Caulerpa racemosa (Cr) has provided anti-inflammatory and antinociceptive activity. This study evaluated these effects on a TMJ hypernociception model. Wistar rats (180 - 250 g) were pre-treated (i.v.) with Cr at 0.01, 0.1, or 1 mg/kg or vehicle 30 min before formalin (1.5%/50 µL, i.art.), capsaicin (1.5%/20 µL, i.art.), or serotonin (225 µg/50 µL, i.art.) in the TMJ, and nociceptive behaviors were measured for 45 or 30 min upon inflammatory stimuli. Inflammatory parameters vascular permeability assay, TNF-α, and IL-1ß by ELISA, protein expression of adhesion molecules ICAM-1 and CD55 by Western blot were assessed. The involvement of heme oxygenase-1 (HO-1) and nitric oxide (NO) pathways were assessed by pharmacological inhibition. Cr (1 mg/kg) reduced nociceptive behavior, plasmatic extravasation, TNF-α, and IL-1ß levels, as well as ICAM-1 and CD55 expression in periarticular tissues. Cr antinociceptive effect was not prevented by aminoguanidine, but ZnPP-IX did reduce its antinociceptive effect. Therefore, Cr antinociceptive and anti-inflammatory effects in this experimental model of hypernociception depended on the HO-1 pathway integrity, as well as reducing peripheral inflammatory events, e.g., TNF-α and IL-1ß cytokines levels, ICAM-1 and CD55 expression.

Qualidade de vida de idosos com doenças crônicas acompanhados pela Estratégia Saúde da Família / Quality of life of elderly people with chronic diseases monitored by the family health strategy

Objetivo: Descrever a qualidade de vida de idosos com doenças crônicas, acompanhados pela Estratégia Saúde da Família. Método: Estudo descritivo e transversal, realizado com 90 idosos acompanhados pelas equipes de saúde da família de um município do Ceará, Brasil, entre outubro e novembro de 2017. Os dados foram coletados nos domicílios dos idosos utilizando um questionário do perfil social e saúde e instrumentos validados sobre qualidade de vida de idosos, WHOQOL-BREF e WHOQOL-OLD. Para análise dos dados empregou-se estatística descritiva. Resultados: A autoavaliação da qualidade de vida obteve média de 48,75. Verificou-se maior escore para a qualidade de vida de idosos nos domínios das relações sociais (57,98) e intimidade (53,04). Houve menor escore para o domínio "atividades passadas", presentes e futuras (37,23) e para o domínio meio ambiente (45,45), onde as facetas de maiores prejuízos foram recursos financeiros (35,00) e recreação e lazer (26,07). Conclusão: Idosos com doenças crônicas alcançam níveis medianos de qualidade de vida, sendo as relações sociais e intimidade os domínios que se apresentam com níveis mais satisfatórios, necessitando melhorar aspectos relacionados à assistência social e de saúde mental e física dos idosos. (AU)

Objective: To describe the quality of life of elderly people with chronic diseases monitored by the Family Health Strategy. Methods: A descriptive and cross-sectional study, carried out with 90 elderly people monitored by health teams in the city of Ceará, Brazil, between October and November 2017. Data were collected in the homes of the elderly using a questionnaire of social and health profile and validated instruments on quality of life of the elderly, WHOQOL-BREF and WHOQOL-OLD. For data analysis, descriptive statistics is used. Results: An average self-assessment of quality of life of 48.75. There was a higher score in the domain of social relations (57.98) and a lower score in the domain of the environment (45.45) where the facets that point the most damage to the quality of life of the elderly were financial resources (35.00) and recreation and leisure (26.07). The intimacy domain used the highest score (53.04) and the past, present and future activities lowest score (37.23) domain by WHOQOL-OLD. Conclusion: Elderly people with chronic diseases reached average levels of quality of life, with social relationships and domains of intimacy presenting more satisfactory levels, needing to improve aspects related to health care and usage practices. (AU)

Objetivo: Describir la calidad de vida de las personas mayores con enfermedades crónicas monitoreadas por la Estrategia de Salud Familiar. Método: Estudio descriptivo y transversal, realizado con 90 personas mayores monitoreadas por equipos de salud en la ciudad de Ceará, Brasil, entre octubre y noviembre de 2017. Los datos fueron recolectados en los hogares de ancianos utilizando un cuestionario de perfil social y de salud e instrumentos validados en calidad de vida de los ancianos, WHOQOL-BREF y WHOQOL-OLD. Para el análisis de datos, se utiliza estadística descriptiva. Resultados: Una autoevaluación promedio de calidad de vida de 48.75. Hubo un puntaje más alto en el dominio de las relaciones sociales (57.98) y un puntaje más bajo en el dominio del medio ambiente (45.45) donde las facetas que señalaron el mayor daño a la calidad de vida de los ancianos fueron los recursos financieros (35.00) y recreación y ocio (26.07). El dominio de la intimidad utilizó el puntaje más alto (53.04) y el dominio del puntaje más bajo de las actividades pasadas, presentes y futuras (37.23) por WHOQOL-OLD. Conclusión: Las personas mayores con enfermedades crónicas alcanzaron niveles promedio de calidad de vida, con relaciones sociales y dominios de intimidad que presentan niveles más satisfactorios, que necesitan mejorar aspectos relacionados con la atención médica y las prácticas de uso. (AU)

Sexuality in old age: knowledge/attitude of nurses of Family Health Strategy. / Sexualidade de idosos: conhecimento/atitude de enfermeiros da Estratégia Saúde da Família.

OBJECTIVE: To evaluate the knowledge and attitudes of Family Health Strategy nurses regarding sexuality in old age. METHOD: Cross-sectional, exploratory, descriptive, quantitative study with nurses of the Family Health Strategy of the city of Sobral-CE. The instruments used were a socio-demographic and professional questionnaire and the Aging Sexual Knowledge and Attitudes Scale, Brazilian version. Data were processed in GraphPad Prism 5®, and the significance level was set at p<0.05. RESULTS: The participants were 56 nurses, most of them female, young adults, who reported they were able to provide orientations about sexuality. The mean score in the knowledge dimension was 29.95 (SD=2.21), in a range of 20 to 60; the mean score in the attitude dimension was 27.14 (SD=2.19), in a range of eight to 40. Participants who declared receiving permanent health education and conducting health education on sexuality presented a significantly favorable knowledge, but no statistically significant attitude. CONCLUSION: Nurses have adequate knowledge about sexuality in old age, but still present conservative attitudes. Investing in processes of continuing health education can improve the knowledge and practice of nurses.

Sexualidade de idosos: conhecimento/atitude de enfermeiros da Estratégia Saúde da Família / Sexualidad de las personas mayores: conocimiento/actitud de enfermeros de la Estrategia Salud de la Familia / Sexuality in old age: knowledge/attitude of nurses of Family Health Strategy

RESUMO Objetivo Avaliar o conhecimento e a atitude dos enfermeiros da Estratégia Saúde da Família sobre sexualidade na velhice. Método Estudo transversal, exploratório-descritivo, de abordagem quantitativa, com enfermeiros da Estratégia Saúde da Família do município de Sobral-CE. Utilizou-se de questionário sociodemográfico e profissional e da Aging Sexual Knowledge And Attitudes Scale , versão brasileira. Os dados foram processados no GraphPad Prism 5®, e estabelecido nível de significância p<0,05. Resultados Participaram 56 enfermeiros, a maioria do sexo feminino, jovens adultas, que se autodeclaram saber orientar sobre sexualidade. A média dos escores do conhecimento foi de 29,95 (DP=2,21), em uma variação de 20 a 60; já para a atitude, a média foi de 27,14 (DP=2,19) na escala, que varia de oito a 40. Os participantes que declararam receber educação permanente em saúde e realizar educação em saúde sobre sexualidade detêm um conhecimento significativamente favorável, mas não foi encontrada atitude significante. Conclusão Os enfermeiros possuem conhecimento adequado sobre a sexualidade na velhice, mas difundem ainda atitudes conservadoras. Investir em processos de educação permanente em saúde pode melhorar o saber e fazer dos enfermeiros.

RESUMEN Objetivo Evaluar el conocimiento y la actitud de enfermeros de la Estrategia Salud de la Familia acerca de la sexualidad en la vejez. Método Estudio transversal, exploratorio y descriptivo, de abordaje cuantitativo, con enfermeros de la Estrategia Salud de la Familia del municipio de Sobral-CE. Se utilizó cuestionario sociodemográfico y profesional y la Aging Sexual Knowledge And Attitudes Scale, versión brasileña. Los datos fueron procesados en el GraphPad Prism 5®, y fue establecido nivel de significación p<0,05. Resultados Participaron 56 enfermeros, la mayoría del sexo femenino, jóvenes adultas, quienes se autodeclararon saber orientar acerca de la sexualidad. El promedio de los scores del conocimiento fue de 29,95 (DP=2,21), en una variación de 20 a 60; ya para la actitud, el promedio fue de 27,14 (DP=2,19) en la escala, que varía de ocho a 40. Los participantes que manifestaron recibir educación permanente en salud y realizar educación sanitaria acerca de la sexualidad detienen un conocimiento significativamente favorable, pero no fue encontrada actitud significativa. Conclusión Los enfermeros tienen conocimiento adecuado sobre sexualidad en la vejez, pero todavía difunden actitudes conservadoras. Invertir en procesos de educación permanente en salud puede mejorar el saber y el hacer de los enfermeros.

ABSTRACT Objective To evaluate the knowledge and attitudes of Family Health Strategy nurses regarding sexuality in old age. Method Cross-sectional, exploratory, descriptive, quantitative study with nurses of the Family Health Strategy of the city of Sobral-CE. The instruments used were a socio-demographic and professional questionnaire and the Aging Sexual Knowledge and Attitudes Scale, Brazilian version. Data were processed in GraphPad Prism 5®, and the significance level was set at p<0.05. Results The participants were 56 nurses, most of them female, young adults, who reported they were able to provide orientations about sexuality. The mean score in the knowledge dimension was 29.95 (SD=2.21), in a range of 20 to 60; the mean score in the attitude dimension was 27.14 (SD=2.19), in a range of eight to 40. Participants who declared receiving permanent health education and conducting health education on sexuality presented a significantly favorable knowledge, but no statistically significant attitude. Conclusion Nurses have adequate knowledge about sexuality in old age, but still present conservative attitudes. Investing in processes of continuing health education can improve the knowledge and practice of nurses.

A lectin fraction from green seaweed Caulerpa cupressoides inhibits inflammatory nociception in the temporomandibular joint of rats dependent from peripheral mechanisms.

Temporomandibular disorders are the second most common cause of orofacial pain mediated by inflammatory compounds, which in many cases leads to chronic orofacial pain. This study assessed the antinociceptive and anti-inflammatory effects of a lectin from the green seaweed Caulerpa cupressoides (CcL) on hypernociception inflammatory in TMJ of rats and investigated the involvement of different mechanisms. Rats received i.v. CcL 30 min prior to injection of flogistic agentes or 0.9% saline into the left TMJ. Pretreatment with CcL (0. 1; 1 or 10 mg/kg) promoted a reduction (p < 0.05) of inflammatory hypernociception induced by 1.5% Formalin along with inhibition of inflammatory plasma extravasation, cytokines levels, ciclooxigenase-2, and intercellular adhesion molecule (ICAM-1). CcL was able to inhibit the nociceptive response induced by 1.5% Capsaicin, suggesting that CcL has an antinociceptive effect, acting directly on the primary nociceptive neurons. CcL also inhibited the nociceptive response induced by Carrageenan (100 µg/TMJ) or Serotonin (5-HT) (225 µg/TMJ). In conclusion, the results demonstrate that administration of CcL has a potential antinociceptive and anti-inflammatory effect, with a mechanism that is partially dependent on TNF-α, IL-1ß, COX-2 and ICAM-1 inhibition and independently from the cannabinoide and opioid system and NO/cGMP/PKG/K+ATP channel pathway.

The efficacy of a lectin from Abelmoschus Esculentus depends on central opioid receptor activation to reduce temporomandibular joint hypernociception in rats.

Abelmoschus esculentus is largely cultivated in Northeastern Brazil for medicinal purposes, e.g. inflammatory conditions. This study aimed to evaluate the efficacy of Abelmoschus esculentus lectin (AEL) in reducing formalin-induced temporomandibular joint inflammatory hypernociception in rats. The behavioral experiments were performed in male Wistar rats (180-240 g). Rats were pre-treated (i.v.) with AEL (0.001, 0.01 or 0.1 mg/kg) 30 min before formalin injection (i.art.). To analyze the possible effect of opioid pathways on AEL efficacy, animals were pre-treated with naloxone or CTOP (µ opioid receptor antagonist), naltrindole (δ opioid receptor antagonist) or nor-binaltorphimine (κ opioid receptor antagonist) (i.t.) 15 min before AEL administration followed by intra-TMJ injection of 1.5% formalin. Animals were monitored for a 45-min observation period. TMJ tissue, trigeminal ganglion, and subnucleus caudalis were collected for TNF-α dosage (ELISA). In addition, the vascular permeability was evaluated by Evans Blue extravasation. AEL significantly reduced formalin-induced TMJ inflammatory hypernociception and decreased Evans blue extravasation. It decreased TNF-α levels in the TMJ tissue, trigeminal ganglion, and subnucleus caudalis. AEL antinociceptive effects were not observed in the presence of naltrindole or nor-binaltorphimine, suggesting that AEL efficacy depends on TNF-α inhibition and the activation of δ and κ opioid receptors. AEL has provided prominent analgesic and anti-inflammatory effects in this pre-clinical model of TMJ, supporting its possible use as a pharmacological tool for the management of painful conditions.

Tocoyena sellowiana extract decreases bone loss in an experimental model of periodontitis in rats: Putative role for cyclooxygenase-2 and IL-1? inhibition.

Tocoyena sellowiana (Cham. & Schltdl.) K.Schum is one of the most important families of Brazilian medicinal plants. This study aimed to evaluate the effect of Tocoyena sellowiana (Cham. & Schltdl.) K.Schum ethanolic extract in a pre-clinical trial of periodontitis and to investigate possible mechanisms underlying such effects. Periodontitis was induced in Wistar rats by placing a nylon thread ligature around second upper left molars for 11 days. Rats received (per os) Tocoyena sellowiana (0.1, 1 or 10?mg?kg) or vehicle 1?h before ligature and daily until day 11. Macroscopic, histopathological, and COX-2 immunohistochemical analyses were performed to evaluate the periodontium. The gingival tissue was used to quantify the myeloperoxidase (MPO) activity and interleukin (IL)-1? levels by ELISA. Blood samples were collected to evaluate bone-specific alkaline phosphatase (BALP), the dosage of creatinine, aspartate and alanine transaminases. The liver, kidneys, spleen, and body mass variations were also evaluated. Tocoyena sellowiana decreased bone loss, reduced MPO, IL-1? levels as well as COX-2 immunostaining, and increased BALP activity. Moreover, Tocoyena sellowiana did not alter organs nor body weight. Tocoyena sellowiana reduced bone loss in rats and its efficacy was at least partially dependent upon both IL-1? and cyclooxygenase-2 inhibition.

Antinociceptive, anti-inflammatory and toxicological evaluation of semi-synthetic molecules obtained from a benzyl-isothiocyanate isolated from Moringa oleifera Lam. in a temporomandibular joint inflammatory hypernociception model in rats.

Inflammation is a key component of many clinical conditions that affect the temporomandibular joint (TMJ) and Moringa oleifera Lam. has been used to treat inflammatory diseases. Here, we evaluated the toxicological effects on mice of a naturally-occurring isothiocyanate from M. oleifera and its seven analogue molecules. Further, the anti-nociceptive and anti-inflammatory effects on a rat model of TMJ inflammatory hypernociception were assessed. The systemic toxicological profile was determined in mice over a 14-day period: MC-1 1 µg/kg; MC-D1 1 µg/kg, MC-D3 100 µg/kg, MC-D6 1 µg/kg, MC-D7 1 µg/kg, MC-D8 1 µg/kg, MC-D9 10 µg/kg, and MC-H 1 µg/kg. The safest molecules were assayed for anti-nociceptive efficacy in the formalin (1.5%, 50 µL) and serotonin (255 mg) induced TMJ inflammatory hypernociception tests. The anti-inflammatory effect was evaluated through the vascular permeability assay using Evans blue. Further, the rota-rod test evaluated any motor impairment. Among the tested molecules, MC-D7, MC-D9, and MC-H were not toxic at the survival rate test, biochemical, and hystological analysis. They reduced the formalin-induced TMJ inflammatory hypernociception, but only MC-H decreased the serotonin-induced TMJ inflammation, suggesting an adrenergic receptor-dependent effect. They diminished the plasmatic extravasation, showing anti-inflammatory activity. At the rota-rod test, no difference was observed in comparison with control groups, reinforcing the hypothesis of anti-nociceptive effetc without motor impairment in animals. The analogues MC-D7, MC-D9, and MC-H were safe at the tested doses and efficient in reducing the formalin-induced TMJ hypernociception in rats. Our next steps include determining their mechanisms of anti-nociceptive action.

Heme oxygenase-1/biliverdin/carbon monoxide pathway downregulates hypernociception in rats by a mechanism dependent on cGMP/ATP-sensitive K+ channels.

OBJECTIVE AND DESIGN: To investigate the role of heme oxygenase-1 (HO-1), carbon monoxide (CO), and biliverdin (BVD) in the zymosan-induced TMJ arthritis in rats. MATERIALS AND METHODS: Mechanical threshold was assessed before and 4 h after TMJ arthritis induction in rats. Cell influx, myeloperoxidase activity, and histological changes were measured in the TMJ lavages and tissues. Trigeminal ganglion and periarticular tissues were used for HO-1, TNF-α, and IL-1ß mRNA time course expression and immunohistochemical analyses. Hemin (0.1, 0.3, or 1 mg kg-1), DMDC (0.025, 0.25, or 2.5 µmol kg-1), biliverdin (1, 3, or 10 mg kg-1), or ZnPP-IX (1, 3 or 9 mg kg-1) were injected (s.c.) 60 min before zymosan. ODQ (12.5 µmol kg-1; s.c.) or glibenclamide (10 mg kg-1; i.p.) was administered 1 h and 30 min prior to DMDC (2.5 µmol kg-1; s.c), respectively. RESULTS: Hemin (1 mg kg-1), DMDC (2.5 µmol kg-1), and BVD (10 mg kg-1) reduced hypernociception and leukocyte migration, which ZnPP (3 mg kg-1) enhanced. The effects of DMDC were counteracted by ODQ and glibenclamide. The HO-1, TNF-α, and IL-1ß mRNA expression and immunolabelling increased. CONCLUSIONS: HO-1/BVD/CO pathway activation provides anti-nociceptive and anti-inflammatory effects on the zymosan-induced TMJ hypernociception in rats.

Anti-inflammatory and antiresorptive effects of Calendula officinalis on inflammatory bone loss in rats.

OBJECTIVE: The aim of this work was to evaluate the anti-inflammatory and antiresorptive effects of Calendula officinalis (CLO) on alveolar bone loss (ABL) in rats. MATERIAL AND METHODS: Male Wistar rats were subjected to ABL by ligature with nylon thread around the second upper left molar. The contralateral hemimaxillae were used as control. Rats received saline solution (SAL) or CLO (10, 30, or 90 mg/kg) 30 min before ligature and daily until the 11th day. The maxillae were removed and prepared for macroscopic, radiographic, micro-tomographic, histopathologic, histometric analysis, and immunohistochemical localization of receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). The gingival tissues were used to quantify the myeloperoxidase (MPO) activity, tumor necrosis factor-alpha (TNF-α), and interleukin-1ß (IL-1ß) concentrations by ELISA. Blood samples were collected for leukogram and to evaluate the bone-specific alkaline phosphatase (BALP) activity and serum levels of aspartate and alanine transaminases (AST/ALT). RESULTS: The bone loss induced by 11 days of ligature induced bone loss, reduced levels of BALP, leukocyte infiltration, increased MPO activity, gingival concentrations of TNF-α and IL-1ß, and RANKL while reduced OPG immunoexpressions in the periodontal tissue and leukocytosis. Of the CLO, 90 mg/kg reduced bone loss, neutrophilia, the levels of pro-inflammatory mediators, and RANKL expression, while it increased OPG immunopositive cells and BALP serum levels, when compared to SAL. CLO did not affect either kidney or liver function, indicated by serum AST/ALT levels. CONCLUSION: The present data suggests that CLO reduced inflammatory bone resorption in experimental periodontitis, which may be mediated by its anti-inflammatory properties and its effects on bone metabolism. CLINICAL RELEVANCE: CLO can be a potential therapeutical adjuvant in the treatment of periodontitis.

Anti-inflammatory and anti-nociceptive effects of strontium ranelate on the zymosan-induced temporomandibular joint inflammatory hypernociception in rats depend on TNF-α inhibition.

BACKGROUND: Temporomandibular joint (TMJ) disorders show inflammatory components, heavily impacting on quality of life. Strontium ranelate has previously shown anti-inflammatory and antinociceptive effects on other experimental inflammatory pain models. Thus, we aim to investigate the strontium ranelate efficacy in reducing the zymosan-induced inflammatory hypernociception in the TMJ of rats by evaluating the TNF-α, IL-1ß, and hemeoxygenase-1 (HO-1) involvement. METHODS: Wistar rats were treated with strontium ranelate (0.5, 5 or 50 mg/kg, per os) 1 h before zymosan injection (iart). Mechanical threshold was assessed by Von Frey test and synovial lavage was collected for leukocyte counting and myeloperoxidase measurement, joint tissue and trigeminal ganglion were excised for histopathological analysis (H&E) and TNF-α/IL-1ß levels dosage (ELISA). Moreover, rats were pre-treated with ZnPP-IX (3 mg/kg, sc), a specific HO-1 inhibitor, before strontium ranelate administration (0.5 mg/kg, per os), and Evans Blue (5 mg/kg, iv) was administered to assess plasma extravasation. Pre-treatment with indomethacin (5 mg/kg, sc) was used as positive control while the sham group received 0.9% sterile saline (per os and iart). RESULTS: Strontium ranelate did not reduce leukocyte counting, myeloperoxidase activity, Evans Blue extravasation, IL-1ß levels, and TNF-α/IL-1ß immunolabeling; but it increased the nociceptive threshold and reduced TNF-α levels. Additionally, HO-1 inhibition did not change the strontium ranelate effects. CONCLUSION: Strontium ranelate may achieve its antinociceptive effects through the reduction of TNF-α levels in the trigeminal ganglion, but not suppressing IL-1ß expression nor inducing the HO-1 pathway.

Mechanisms involved in antinociception induced by a polysulfated fraction from seaweed Gracilaria cornea in the temporomandibular joint of rats.

Temporomandibular disorder is a common clinical condition involving pain in the temporomandibular joint (TMJ) region. This study assessed the antinociceptive effects of a polysulfated fraction from the red seaweed Gracilaria cornea (Gc-FI) on the formalin-induced TMJ hypernociception in rats and investigated the involvement of different mechanisms. Male Wistar rats were pretreated with injection (sc) of saline or Gc-FI 1h before intra- TMJ injection of formalin to evaluate the nociception. The results showed that pretreatment with Gc-FI significantly reduced formalin-induced nociceptive behavior. Moreover, the antinociceptive effect of the Gc-FI was blocked by naloxone (a non-selective opioid antagonist), suggesting the involvement of opioids selective receptors. Thus, the pretreatment with selective opioids receptors antagonists, reversed the antinociceptive effect of the Gc-FI in the TMJ. The Gc-FI antinociceptive effect depends on the nitric oxide/cyclic GMP/protein kinase G/ATP-sensitive potassium channel (NO/cGMP/PKG/K+ATP) pathway because it was prevented by pretreatment with inhibitors of nitric oxide synthase, guanylate cyclase enzyme, PKG and a K+ATP blocker. In addition, after inhibition with a specific heme oxygenase-1 (HO-1) inhibitor, the antinociceptive effect of the Gc-FI was not observed. Collectively, these data suggest that the antinociceptive effect induced by Gc-FI is mediated by µ/δ/κ-opioid receptors and by activation NO/cGMP/PKG/K+ATP channel pathway, besides of HO-1.

Role of central opioid on the antinociceptive effect of sulfated polysaccharide from the red seaweed Solieria filiformis in induced temporomandibular joint pain.

This study aimed to investigate the effect of sulfated polysaccharide from red seaweed Solieria filiformis (Fraction F II) in the inflammatory hypernociception in the temporomandibular joint (TMJ) of rats. Male Wistar rats were pretreated (30min) with a subcutaneous injection (s.c.) of vehicle or FII (0.03, 0.3 or 3.0mg/kg) followed by intra-TMJ injection of 1.5% Formalin or 5-hydroxytryptamine (5-HT, 225µg/TMJ). In other set of experiments rats were pretreated (15min) with an intrathecal injection of the non-selective opioid receptors Naloxone, or µ-opioid receptor antagonist CTOP, or δ-opioid receptor Naltridole hydrochloride, or κ-opioid receptor antagonist Nor-Binaltorphimine (Nor-BNI) followed by injection of FII (s.c.). After 30min, the animals were treated with an intra-TMJ injection of 1.5% formalin. After TMJ treatment, behavioral nociception response was evaluated for a 45-min observation period, animals were terminally anesthetized and periarticular tissue, trigeminal ganglion and subnucleus caudalis (SC) were collected plasma extravasation and ELISA analysis. Pretreatment with F II significantly reduced formalin- and serotonin-induced TMJ nociception, inhibit the plasma extravasation and inflammatory cytokines release induced by 1.5% formalin in the TMJ. Pretreatment with intrathecal injection of Naloxone, CTOP, Naltridole or Nor-BNI blocked the antinociceptive effect of F II in the 1.5% formalin-induced TMJ nociception. In addition, F II was able to significantly increase the ß-endorphin release in the subnucleus caudalis. The results suggest that F II has a potential antinociceptive and anti-inflammatory effect in the TMJ mediated by activation of opioid receptors in the subnucleus caudalis and inhibition of the release of inflammatory mediators in the periarticular tissue.

Lectin from Abelmoschus esculentus reduces zymosan-induced temporomandibular joint inflammatory hypernociception in rats via heme oxygenase-1 pathway integrity and tnf-α and il-1ß suppression.

Temporomandibular joint (TMJ) disorders show inflammatory components, heavily impacting on quality of life. Abelmoschus esculentus is largely cultivated in Northeastern Brazil for medicinal purposes, having it shown anti-inflammatory activity. We evaluated A. esculentus lectin (AEL) efficacy in reducing zymosan-induced temporomandibular joint inflammatory hypernociception in rats along with the mechanism of action through which it exerts anti-inflammatory activity. Animals were pre-treated with AEL (0.01, 0.1 or 1mg/kg) before zymosan (Zy) injection in the TMJ to determine anti-inflammatory activity. To analyse the possible effect of the hemeoxygenase-1 (HO-1) and the nitric oxide (NO) pathways on AEL efficacy, animals were pre-treated with ZnPP-IX (3mg/kg), a specific HO-1 inhibitor, or aminoguanidine (30mg/kg), a selective iNOS inhibitor, before AEL administration. Von Frey test evaluated inflammatory hypernociception, synovial fluid collection was performed to determine leukocyte counting and myeloperoxidase (MPO) activity 6h after Zy injection, and Evans Blue extravasation determined vascular permeability. TMJ tissue was collected for histopathological analysis (H&E) and immunohistochemistry (TNF-α, IL-1ß, HO-1). In addition, TMJ tissue and trigeminal ganglion collection was performed for TNF-α and IL-1ß dosage (ELISA). AEL increased inflammatory nociceptive threshold, reduced leukocyte influx along with MPO activity, leukocyte influx into the synovial membrane, and Evans Blue extravasation. It promoted HO-1 overexpression whilst decreased TNF-α and IL-1ß expression in the TMJ tissue. AEL reduced TNF-α and IL-1ß levels in TMJ tissue and trigeminal ganglion. AEL effects, however, were not observed in the presence of ZnPP-IX. These findings suggest that AEL efficacy depends on TNF-α/IL-1ß inhibition and HO-1 pathway integrity.

Atendimento à Lei Arouca no Ensino de Farmacologia no Curso de Medicina, UFC, Sobral / Compliance with Arouca’s Law in Teaching Pharmacology at the UFC Medical School, Sobral

RESUMO De acordo com a Lei Arouca (Lei n° 11.794 – 2008), que estabelece critérios para “a criação e a utilização de animais em atividades de ensino e pesquisa científica, em todo o território nacional”, tem-se que, “sempre que possível, as práticas de ensino deverão ser fotografadas, filmadas ou gravadas, de forma a permitir sua reprodução para ilustração de práticas futuras, evitando-se a repetição desnecessária de procedimentos didáticos com animais”. O objetivo deste estudo foi relatar o desenvolvimento de um software (Pharmasoftware®) como método alternativo de ensino em Farmacologia no curso de Medicina da Universidade Federal do Ceará (UFC), Campus Sobral. Para a certificação, foram utilizados dois grupos de alunos, um em aula prática convencional e outro com o Pharmasoftware®. A análise dos resultados revelou que ambas as atividades foram igualmente eficazes em auxiliar na consolidação do tema Vias de Administração dos Fármacos, evidenciando, assim, o Pharmasoftware® como uma ferramenta capaz de auxiliar no ensino da Farmacologia e, ainda, de fomentar a implantação de outros métodos alternativos de ensino de Farmacologia nas Instituições de Ensino Superior.

ABSTRACT The Brazilian Law on scientific and teaching use of animals, Arouca’s Law (n° 11.794 – 2008), clearly states that animals should not be used if an alternative method is available. The aim of this study was to describe the development of a software (Pharmasoftware®) as an alternative method of teaching in pharmacology at a Medical School (Federal University of Ceara, Campus Sobral). To support the development and validation of the Pharmasoftware® as an alternative method, students were allocated into two groups: one attending a conventional practical class and the other employing Pharmasoftware®. The results showed that two activities were equally effective in improving the students’ knowledge of drug administration. Pharmasoftware® may therefore represent a tool capable of supporting pharmacology teaching and it also fostering the establishment of alternative methods for teaching Pharmacology in the undergraduate courses.

A lectin from the green seaweed Caulerpa cupressoides reduces mechanical hyper-nociception and inflammation in the rat temporomandibular joint during zymosan-induced arthritis.

Seaweed lectins have been widely investigated as anti-nociceptive and anti-inflammatory agents. This study analyzed the anti-nociceptive and anti-inflammatory responses of a lectin from the green seaweed Caulerpa cupressoides (CcL) on zymosan-induced arthritis of the rat temporomandibular joint (TMJ). Rats received i.v. CcL 30 min prior to injection of zymosan (2mg/art.) or 0.9% saline into the left TMJ. Mechanical hyper-nociception was measured by the electronic von Frey method at baseline and 4h after zymosan injection. Animals were euthanized 6h after zymosan injection and the synovial fluid was collected for leukocyte counting and myeloperoxidase activity assessment. Other animals were treated with ZnPP-IX (3mg/kg; s.c.), a specific heme oxygenase-1 pathway inhibitor, and naloxone (10 µg/art.), a nonselective opioid receptor antagonist. TMJ tissues were excised to perform histopathological and immunohistochemistry analyses. CcL (0.1, 1 or 10mg/kg) significantly reduced zymosan-induced hyper-nociception (81, 83 and 89.5%, respectively) and inhibited the leukocyte influx (77.3, 80.7 and 98.5%, respectively) compared with the zymosan-only group, as confirmed by myeloperoxidase activity; however, treatment with naloxone or ZnPP-IX did not revert the effects of CcL (10mg/kg), suggesting that the naloxone-sensitive opioid and heme oxygenase-1 pathways are not involved. CcL also reduced the leukocyte influx and the expression of IL-1ß and TNF-α in the TMJ, based on histopathological and immunohistochemistry analyses, respectively. Therefore, CcL reduces TMJ hyper-nociception and inflammation with a mechanism that is partially dependent on TNF-α and IL-1ß inhibition. CcL reveals a potentially valuable alternative tool for future studies of TMJ disorders.