Desenvolvimento e validação de métodos analíticos para determinação do cloridrato de epinastina em comprimidos revestidos

O cloridrato de epinastina é um agente não sedativo, antagonista H1 da histamina, utilizado no tratamento da asma ou como anti-histamínico, comercializado no Brasil na forma farmacêutica comprimido revestido com o nome de Talerc ®. Embora, seja um medicamento amplamente comercializado não há referências relativas aos métodos de análise da forma farmacêutica. Este trabalho teve como objetivos o desenvolvimento e a validação de métodos analíticos para o controle de qualidade do cloridrato de epinastina nos comprimidos revestidos, bem como a realização de estudos preliminares de estabilidade térmica e de fotoestabilidade e determinação da cinética de reação do anti-histamínico em estudo.A análise qualitativa do cloridrato de epinastina foi realizada por ressonância magnética nuclear de próto n, determinação da faixa de fusão, espectrofotometria na região do infravermelho e do ultravioleta, cromatografia em camada delgada e por cromatografia a líquido de alta eficiência. Para análise quantitativa, foram validados os métodos por espectrofotometria na região do ultravioleta derivada de primeira ordem e por cromatografia a líquido de alta eficiência. A validação foi efetuada de acordo com as guias de validação disponíveis na literatura. No estudo preliminar de estabilidade térmica utilizou-se a forma farmacêutica submetida a temperatura de 80 °C por 10 dias. A fotoestabilidade foi realizada em câmara de luz ultravioleta de 254 nm por um período de 18 horas. Os métodos qualitativos foram úteis para identificação do cloridrato de epinastina. Os métodos quantitativos propostos foram validados de acordo com os parâmetros analíticos preconizados, podendo ser perfeitamente intercambiáveis. O estudo preliminar de estabilidade térmica não apresentou degradação da amostra nas condições de análise empregadas. O estudo de fotoestabilidade apresentou degradação intensa obedecendo uma cinética de reação de primeira ordem.

Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

Avaliação das atividades anti-inflamatória, anticoagulante e antiproliferativa do inibidor de quimotripsina das sementes de erythrina velutina (EvCI)

Studies indicate that several components were isolated from medicinal plants, which have antibacterial, antifungal, antitumor and anti-inflammatory properties. Sepsis is characterized by a systemic inflammation which leads to the production of inflammatory mediators exacerbated by excessive activation of inflammatory cells and disseminated intravascular coagulation (DIC), in which the human neutrophil elastase plays an important role in its pathogenesis. Several epidemiological studies suggest that components of plants, especially legumes, can play a beneficial role in reducing the incidence of different cancers. A chymotrypsin inhibitor of Kunitz (Varela, 2010) was purified from seeds of Erythrina velutina (Mulungu) by fractionation with ammonium sulfate, affinity chromatography on Trypsin-Sepharose, Chymotrypsin-Sepharose and ion exchange chromatography on Resource Q 1 ml (GE Healthcare) in system FPLC / AKTA. The inhibitor, called EvCI, had a molecular mass of 17 kDa determined by SDS-PAGE. The purified protein was able to inhibit human neutrophil elastase (HNE), with an IC50 of 3.12 nM. The EvCI was able to inhibit both pathways of HNE release stimulated by PAF and fMLP (75.6% and 65% respectively). The inhibitor also inhibited leukocyte migration in septic mice about 87% and prolonged the time of coagulation and inhibition factor Xa. EvCI showed neither hemolytic activity nor cytotoxicity. EvCI showed a selective antiproliferative effect to HepG2 cell lines with IC50 of 0.5 micrograms per milliliter. These results suggest EvCI as a molecule antagonist of PAF / fMLP and a potential use in fighting inflammation related disorders, disseminated intravascular coagulation (DIC) and cancer

Efeito pró-inflamatório de uma lectina purificada da esponja marinha Cliona varians

A galactose and sucrose specific lectin from the marine sponge Cliona varians named CvL was purified by acetone fractionation followed by Sepharose CL 4B affinity chromatography. Models of leukocyte migration in vivo were used to study the inflammatory activity of CvL through of mouse paw oedema and peritonitis. Effect of CvL on peritoneal macrophage activation was analyzed. Effects of corticoids and NSAIDS drugs were also evaluated on peritonitis stimulated by CvL. Results showed that mouse hind-paw oedema induced by sub plantar injections of CvL was dependent dose until 50µg/paw. This CvL dose when administered into mouse peritoneal cavities induced maxima cell migration (9283 cells/µL) at 24 hours after injection. This effect was preferentially inhibited by incubation of CvL with the carbohydrates D-galactose followed by sucrose. Pre-treatment of mice with 3% thioglycolate increases the peritoneal macrophage population 2.3 times, and enhanced the neutrophil migration after 24h CvL injection (75.8%, p<0.001) and no significant effect was observed in presence of fMLP. Finally, Pre-treatment of mice with dexamethason (cytokine antagonist) decreased 65.6%, (p<0.001), with diclofenac (non-selective NSAID) decreased 34.5%, (p<0.001) and Celecoxib (selective NSAID) had no effect on leukocyte migration after submission at peritonitis stimulated by CvL, respectively. Summarizing, data suggest that CvL shows pro-inflammatory activity, inducing neutrophil migration probably by pathway on resident macrophage activation and on chemotaxis mediated by cytokines

Avaliação de efeitos toxicológicos e comportamentais de ginseng panax em ratos

Panax ginseng CA Meyer (Araliaceae) is a herbaceous plant widely used in China, South Korea, Japan and other Asian countries for the treatment of various diseases micro circulatory, cerebrovascular, among others, representing one of the drugs used by older man. It has over 30 biologically active ginsenosides with different pharmacological and behavioral effects and inhibitory effect on the NMDA receptor. The amino acid glycine is a co-agonist of the NMDA receptor, activating this receptor. At the cellular level, ketamine is widely known to be NMDA receptor antagonist. The aim of this study was to evaluate the general activity in the open field, and anxiety in elevated plus maze, mice treated with P. ginseng compared with the action of ketamine and glycine, to better understand the action of this herbal medicine at the NMDA receptor. We used 66 adult male rats were divided into six groups: a positive control, treated for 30 days with water by gavage, who received glycine (500mg/kg; po) on days 7, 14, 21 and 28 of treatment, one hour before of behavioral assessment, a negative control was treated for 30 days with water by gavage received ketamine (5mg/kg, ip) on days 7, 14, 21 and 28 of treatment, one hour prior to behavioral evaluation, three experimental groups, receiving 100, 200 or 300 mg / kg P. ginseng by gavage for 30 days and one group treated solely with white water, and is also administered 1 ml of water by gavage one hour prior to behavioral evaluation. Animal behavior in these three groups was also examined on days 7, 14, 21 and 28 of treatment. On day 30 of treatment, the animals were anesthetized with thiopental (70mg/kg) for blood collection and after euthanasia, withdrawal of various organs. There were no changes in weight and body weight gain and weight reasons in organ / body weight. However the consumption of water and food values showed a significant increase. Serum levels of AST was increased in a dose-dependently in the animals treated with doses of P. ginseng, glycine and ketamine as compared to the blank group. Unlike creatinine levels proved to be decreased in all treated groups when compared with white. However, the level of urea in these groups was reduced and no changes were observed in the ALT parameter. Histopathological examination revealed no changes in cell morphology in different tissues. There were no behavioral changes in the elevated plus maze and few changes were observed in the open field, animals treated with P. ginseng, glycine and ketamine when compared to white. These data suggest that the doses of P. ginseng employed were unable to induce general toxicity in rats treated for 30 days and also shows that the general behavior of mice treated with P. ginseng was slightly different from that observed in animals treated with ketamine and glycine. Finally, the study on the elevated plus maze showed that the extract of P. ginseng showed no anxiolytic or anxiogenic action

Inibição do crescimento de Escherichia coli isolada de Queijo Minas Frescal por Lactobacillus acidophilus

Escherichia coli faz parte da microbiota anaeróbica facultativa normal, sendo também considerada um dos maiores patógenos entéricos predominantes no cólon dos animais e homem. Neste trabalho, realizaram-se ensaios in vitro para avaliar o grau de atividade antagonista de cinco cepas de Lactobacillus acidophilus, com capacidade probiótica sobre Escherichia coli BIA 26 (STEC) isolada de queijo Minas Frescal. Para tanto, foi utilizado o teste de inibição através do método de dupla camada em triplicata para avaliar zonas de inibição de crescimento. Todas as cepas de Lactobacillus mostraram-se capazes de inibir a E. coli, com zonas de inibição variando de 12 a 15mm de diâmetro, sendo que a maioria apresentou 14mm de diâmetro, consideradas como fortes halos de inibição.

Avaliação da esporulação de Bionectria ochroleuca em diferentes meios de culturas

The biological control of the plants diseases, caused by fungi, is carried out using others organisms (predator, parasite or pathogen). Among the possible agents of biocontrol, a fungus has been highlighting as promising and it is known as Clonostachys rosea, asexual form of Bionectria ochroleuca. For that, it is necessary the in vitro production of spores of this fungus. In this study were tested several culture media to select those with better conidia production. The study was conducted at Plant Protection Division, in the Plant Production Department, FCA - UNESP, Botucatu campus, São Paulo state, Brazil. We used the isolated CCR64 (Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)-CNPMA). The means of crops were: BDA; Oats-Agar; Mazeina-Agar; Rice-Agar; V8-5%; V8-10%; V8-20%; TJ-5 %; TJ-10, TJ-20%. The sporulation of the fungus in different culture media was estimated at 8 days after of the incubation. The data were analyzed using method of comparing averages, using the Tukey test, at 5% probability, and the data processed, using (X + 1) 0.5 transformation. All culture media tested were able to produce conidia. It was found that the best culture media for production of conidia of Bionectria ochroleuca is the TJ-5%, followed by TJ-20%, with an sporulation average of 3,5 x 106 conidia / ml.

Análise da função dos músculos do quadril em portadores de síndrome da dor patelofemural

The Patellofemoral pain syndrome is defined as a fore or retro patellar pain and it has multifactorial etiology, where the bad patellar alignment is the most acceptable hypothesis. However proximal factors to the knee, as the debility of the muscles of the hip, have been demonstrated as a contributing factor to the appearing of that syndrome. Purpose: To evaluate if exists a relation between the hip muscles performance and the development of the SDPF. Methods: Thirty women took part in this study. They were divided in two groups; a control group (fifteen asymptomatic subjects) and an experimental group (fifteen subjects with the diagnosis of SDPF). The muscle performance was evaluated in an isokinetic dynamometer, where it was verified the peak torque (PT), PT to body weight, PT time and the agonist/antagonist relation. It was also analyzed the electromyographic activity of the middle gluteus. The data was analyzed by the not paired t test at a significance level of 5%. Results:. Didn t have significant difference to the PT of the abductor muscles (p = 0,46) and lateral rotators of the hip (p = 0,17) between groups. Also didn t have significant difference to the PT values by the body weight, to these muscle groups either (p = 0,10 e p = 0,11, respectively). Didn t have significant difference between the amplitude of the signal (p = 0,05) and the onset of medium gluteus (p = 0,25) between the groups. Conclusion: In the experimental conditions realized, the study didn t demonstrate a relation between performance the hip muscles behavior and the development of the SDPF

Efeitos imediatos da eletroestimulação nervosa transcutânea e crioterapia na espasticidade e na atividade eletromiográfica de sujeitos hemiparéticos

Cerebrovascular accident (CVA) is a term used to characterize an ischemic or hemorrhagic vascular injury, which has got as main clinic manifestations, the motor and reflex function disturbance. In the first stage there is flaccidity and loss of voluntary movements that afterwards is substituted by mass patterns and spasticity. The spasticity brings with itself functional deficits and can generate negative impacts in various motor patterns. The aim of this research was to investigate the hyperreflexia and identify the immediate effects of transcutaneous nervous stimulation (TENS) and cryotherapy in the spasticity and electromyographic activity of hemiparetic subjects. The study is characterized as an almost experimental type, in which were selected, to compose the sample, 16 patients of both sex with CVA sequel. These individuals were evaluated by collecting the amplitude peak to peak and H reflex latency, Motor response (M response) in solear muscle and the electromyography (EMG) of the injured and healthy legs anterior tibial muscles. In the injured limb the evaluations occurred in different days for cryotherapy, TENS and control, in two moments, before and after the interventions. The healthy limb was evaluated one single time to serve as baseline, for comparison with the injured limb. It was used an statistic analysis, the t paired student test to identify the H reflex differences, latency and EMG of the injured and healthy limbs and to compare the results before and after the recourses application. The ANOVA for related samples was used to identify the differences among the recourses used. It was attributed for the statistic tests a significance level of 5%. The amplitude peak to peak of normalized maximum H reflex through the maximum motor response (Hmax/Mmax), showed itself significantly increased in the injured limb (p=0.0245). The H reflex latency was presented reduced in the injured limb (p=0, 0375). The electromyographic activity was showed decreased in the injured limb (p< 0.0001). After the TENS there was a Hmáx/Mmáx ratio decrease (0.60±0.16 versus 0.49.±0.18; P = 0.0006). Nonetheless, Just after the cryotherapy application there was an increase of Hmáx/Mmáx ratio (0.58 ± 0,15 to 0.77 ± 0.13, P=0,0007) and increase of signal latency (30.41 ± 1.87 versus 33.24 ± 2.19; P=0.0001). The electromyographic activity wasn t altered significantly by any resource. It was met statistic significant differences when the Hmáx/Mmáx P<0.0001) ratio and H reflex latency (P<0.0001) were compared between the post TENS, cryotherapy and control. One can conclude that the TENS can be used to spasticity immediate reduction, and that the cryotherapy can increase the hyperreflexia state in spastic patients. Nonetheless, the spasticity decrease or increase didn t provoke lectromyographic activity change in the muscle that is opponent to the spastic one

Efeitos da administração aguda de quetamina sobre as oscilações eletrofisiológicas da região CA1 hipocampal

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Avaliação da desadaptação das selas de próteses parciais removíveis após 1 a 5 anos de uso

The non-adaptation of the removable partial prosthesis (RPP) base to fibromucosal tissue is caused by resorption of residual ridges (RRR). The onset of bone resorption, which occurs after tooth extraction and continues throughout life, is accelerated by local or systemic factors. Aim: Assess the degree of non-adaptation of removable partial prosthesis saddles and the factors that influence it. Methodology: A sectional study was conducted with 81 patients using RPP who had their prostheses installed between 2003 and 2007 (1 to 5 years of use) at the Faculty of Dentistry of the Universidade Federal do Rio Grande do Norte (UFRN). After anamnese and clinical examination, a cast was made with polyether-based material, using the base of the prosthesis to make the impression. The base of the saddle was loaded with the casting material and positioned in the mouth, applying pressure on the supports. After polymerization, the material was removed from the saddle and measurements were taken at 3 different points using a pachymeter. Results: The non-adaptation of the saddle increased significantly with years of use (p = 0.005). The tooth-tissue supported prostheses obtained higher mean non-adaptation values than those of tooth supported prostheses (p < 0.001). Flaccid mucosa showed the worst non-adaptation results, which were statistically different from resilient mucosa (p < 0.001). The greater the extension of the saddle, the greater the non-adaptation (p < 0.001). The natural tooth antagonistic arch yielded better results than did RPP and total prosthesis (p < 0.001). Saddle non-adaptation at the free end was less near the pillar tooth and greater in the more posterior region (p < 0.001). When adaptation of the supports to the niches was poor, greater saddle non-adaptation occurred than when it was good or fair (p < 0.001). Saddles located in the posterior region of the arch had greater non-adaptation than those in the anterior region (p = 0.023). Conclusion: The mean non-adaptation of the saddle to the residual ridges was 0.27 mm. It can be concluded that, even with the use of RPP, bone height reduction was slight within the 1-5-year period of use. The following are factors that influence adaptation of the RPP saddle base: years of use, age, force transmission path to the alveolar bone, location of the toothless area, antagonistic arch, type of mucosa, adaptation of supports to the niche and extension of the saddle

Influência do ciclo estral no efeito do diazepam na ansiedade e memória de ratas

Memory and anxiety are related phenomena. Several evidences suggest that anxiety is fundamental for learnining and may facilitate or impair the memory formation process depending of the context. The majority of animal studies of anxiety and fear use only males as experimental subjects, while studies with females are rare in the literature. However, the prevalence in phobic and anxiety disorders is greater in women than in men. Moreover, it is known that gender maybe influence benzodiazepine effects, the classic drugs used for anxiety disorders treatment. In this respect, to further investigate if fear/anxiety aspects related to learning in female subjects would contribute to the study of phobic and anxiety disorders and their relationship with learning/memory processes, the present work investigates (a) the effects of benzodiazepine diazepam on female rats performance in a aversive memory task that assess concomitantly anxiety/emotionality, as the interaction between both; (b) the influence of estrous cycle phases of female rats on diazepam effects at aversive memory and anxiety/emotionality, and the interaction between both and (c) the role of hormonal fluctuations during estrous cycle phases in absence of diazepam effects in proestrus, because female rats in this phase received or not mifepristone, the antagonist of progesterone receptor, previously to the diazepam treatment. For this purpose, the plus maze discriminative avoidance task, previously validated for studies of anxiety concomitantly to learning/memory, was used. The apparatus employed is an adaptation of a conventional plus maze, with two opens arms and two closed arms, one of which presenting aversive stimulation (noise and light). The parameters used were: time in non-aversive arm compared to time in aversive and percentage of time in aversive arm on several temporal divisions, in order to evaluate memory; percentage of time in open arms, risk assessment, head dipping and end exploring to evaluate anxiety ; and distance traveled for locomotion. In experiment I, we found anxiolytic effect of diazepam only for 4 mg/kg dose, however the amnestic effect appear at a dose of 2 mg/kg. In second experiment, rats were divided in groups according estrous cycle phase (metaestrus/diestrus, proestrus e estrus). In this experiment, when we considered estrous cycle phase or diazepam treatment, the results did not demonstrate any differences in anxiety/emotionality parameters. The amnestic effects of diazepam occur in female rats in metestrus/diestrus and estrus and is absent in proestrous rats. Proestrous female rats that received mifepristone exhibited the amnestic effect of diazepam and also anxiolytic effects, that it was not previously observed in this dose. The results have demonstrated dissociation of anxiolytic and amnestic diazepam effects, not previously observed in males; the absence of amnestic effect of diazepam in proestrous phase; and the possible role of progesterone in aversive memory over diazepam effect, because the mifepristone, associated with diazepam, caused amnestic effect in proestrus

Alterações na evocação de uma memória aversiva ao liongo do ciclo estral de ratas: influências da transmissão gabaérgica na amígdala

GABAergic neurotransmission has been implicated in many aspects of learning and memory, as well as mood and anxiety disorders. The amygdala has been one of the major focuses in this area, given its essential role in modulating emotionally relevant memories. However, studies with male subjects are still predominant in the field. Here we investigated the consequences for an aversive memory of enhancing or decreasing GABAergic transmission in the basolateral nucleus of the amygdala (BLA). Wistar female rats were trained in the plus-maze discriminative avoidance task, in which they had to learn to avoid one of the enclosed arms where an aversive stimulus consisting of a bright light and a loud noise was given (day 1). Fifteen minutes before the test session (day 2) animals received 0,2 μL infusions of either saline solution, the GABAergic agonist muscimol (0,05 mg/ml), or the GABAergic antagonist bicuculine (0,025 mg/ml) bilaterally intra-BLA. On the test day, females in proestrous or estrous presented adequate retrieval and did not extinguish the task, while females in metestrous or diestrous presented impaired retrieval. In the first group, muscimol infusion impaired retrieval and bicuculline had no effect, suggesting naturally low levels of GABAergic transmission in the BLA of proestrous and estrous females. In the second group, muscimol infusion had no effect and bicuculline reversed retrieval impairment, suggesting naturally high levels of GABAergic transmission in the BLA of metestrous and diestous females. Additionally, proestrous and estrous females presented higher anxiety levels compared to metestrous and diestrous females, which could explain better performance of this group. On the other hand, BLA GABAergic system did not interfere with the innate fear response because drug infusions had no effect in anxiety. Thus, retrieval alterations caused by the GABAergic drugs were probably related specifically to memory processes

Remodelação miocárdica após infarto agudo do miocárdio experimental em ratos: efeito do bloqueio do sistema renina angiotensina aldosterona

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Impacto de herbicidas em isolados de Trichoderma spp.

O uso de microrganismos é uma alternativa para o controle de doenças em plantas. Todavia, é prudente verificar a interação desse com os demais métodos de controle empregados em determinada cultura. Dessa forma, objetivou-se avaliar a fungitoxicidade dos herbicidas sobre o crescimento e desenvolvimento dos isolados de Trichoderma spp. Utilizou-se o delineamento inteiramente casualizado, em esquema fatorial 6 x 6 x 4, com quatro repetições. O fator A correspondeu aos herbicidas pendimethalin, clomazone, carfentrazone-ethyl, oxadiazon, thiobencarb + propanil e byspiribac-sodium; o fator B, às doses dos herbicidas - 0, 25, 50, 75, 100 e 200% da dose recomendada; e o fator C, aos isolados de Trichoderma spp. AJAM 18, CE 66, TRI 01 e TRI 02. O ensaio foi realizado em condições in vitro; avaliaram-se o crescimento micelial radial (CMR) e a esporulação dos isolados após aplicação dos herbicidas. Observaram-se diferenças de sensibilidade dos isolados para o mesmo produto testado. O oxadiazon reduziu o CMR dos isolados AJAM 18 e TRI 01 em 66 e 35%, respectivamente. No entanto, reduziu apenas 16% do CMR do isolado TRI 02 e não alterou o CMR do isolado CE 66 mesmo em 200% da dose recomendada. Verificaram-se diferentes efeitos dos produtos em cada isolado. A mistura comercial de thiobencarb+propanil foi altamente tóxica aos isolados de Trichoderma spp., com reduções em torno de 85% no CMR e no número de esporos. Por outro lado, o byspiribac-sodium pouco afetou os isolados, apresentando reduções inferiores a 10% no CMR e na esporulação. O carfentrazone-ethyl e byspiribac-sodium demonstraram ser compatíveis com os isolados de Trichoderma spp. estudados.