The essential oil of Hyptis crenata Pohl ex Benth. presents an antiedematogenic effect in mice

Hyptis crenata, commonly known as "salva-do-Marajó", "hortelã-do-campo", and "hortelãzinha", is used in folk medicine in Northeast Brazil as tea or infusion to treat inflammatory diseases. Due to the pharmacological efficacy and the low toxicity of the essential oil of Hyptis crenata (EOHc), we decided to investigate the EOHc antiedematogenic effect in experimental models of inflammation. EOHc was administrated orally at doses of 10-300 mg/kg to male Swiss albino mice. Paw edema was induced by subcutaneous injection in the right hind paw of inflammatory stimuli (carrageenan, dextran, histamine, serotonin, and bradykinin) 60 min after administration of EOHc. EOHc significantly inhibited the induced edema. The inhibitory effect of EOHc on dextran-induced edema extended throughout the experimental time. For the 30, 100, and 300 mg/kg doses of EOHc, the inhibition was of 40.28±1.70, 51.18±2.69, and 59.24±2.13%, respectively. The EOHc inhibitory effect on carrageenan-induced edema started at 10 mg/kg at the second hour (h) and was maintained throughout the observation period. At 30, 100, and 300 mg/kg doses the inhibition started earlier, from 30 min. At the edema peak of 180 min, 56, 76, and 82% inhibition was observed for 30, 100, and 300 mg/kg doses, respectively. Additionally, the effect of EOHc on carrageenan-induced paw edema was influenced by the time of administration. The EOHc also inhibited myeloperoxidase activity. In conclusion, the EOHc showed a potent effect, both preventing and reversing the edema, consistent with its anti-inflammatory use in folk medicine.

Efeito do óleo essencial do Croton nepetifolius Baill na penetração cervical em ovelhas mestiças de Dorper e Santa Inês / Effect of the essential oil of Croton nepetifolius Baill on cervical penetration in crossbred Dorper and Santa Inês ewes

A fim de se avaliar o uso do óleo essencial de Croton nepetifolius Baill (OECn) na penetrabilidade cervical em ovelhas mestiças, 40 ovelhas foram distribuídas ao acaso em quatro grupos (n=10): controle, misoprostol, OECn50 e OECn100 (50 e 100µg do OECn, respectivamente). Após a sincronização do estro, utilizando CIDR e eCG (200UI), a profundidade de penetração da cérvix foi mensurada utilizando-se uma pipeta de inseminação artificial de bovino graduada, no período de zero até 72h após a retirada do CIDR. Os resultados foram expressos em média ± erro-padrão, submetidos à ANOVA seguida do teste de Tukey, enquanto os dados, em porcentagem, foram submetidos aos testes de Fisher ouqui-quadrado. Nenhuma diferença significativa (P>0,05) foi encontrada quanto ao grau de penetrabilidade cervical. Quanto ao tempo de passagem, os grupos misoprostol e OECn100 apresentaram um menor tempo de penetrabilidade às 60h(1,7±0,6 e 1,5±0,6min, respectivamente), quando comparados ao grupo controle (4,1±0,6min), que não diferiu significativamente do grupo OECn50 (2,3±0,6min). Portanto, o óleo essencial de Croton nepetifolius Baill pode ser utilizado para encurtar o tempo de penetrabilidade cervical em ovelhas submetidas à sincronização estral.(AU)

In order to evaluate the use of the essential oil of Croton nepetifolius Baill (EOCn) on cervical penetration in crossbred ewes, 40 ewes were randomly allocated into four groups (n= 10): CONTROL, MISOPROSTOL, EOCn50 and EOCn100 (50 e 100µg of the EOCn, respectively). After estrus synchronization, using CIDR and eCG (200IU), depth of cervical penetration was measured using artificial insemination gun for bovine species which was graduated and used from 0 to 72h after CIDR removal. Results were expressed as mean ± standard error mean, submitted to ANOVA and Tukey test while data in percentage were submitted to Fisher or Chi-Square test. No significant difference (P> 0.05) was observed at grade of cervical penetration. Concerning trespassing time, MISOPROTOL and EOCn100 groups presented a lower trespassing time at 60h (1.7±0.6 and 1.5±0.6min, respectively) than CONTROL group (4.1±0.6min), which did not differ significantly from EOCn50 (2.3±0.6min) group. Therefore, the essential oil of Croton nepetifolius Baill can be used to shorten the cervical penetration time in estrus synchronized ewes.(AU)

Produção e caracterização de protease fibrinolítica de Streptomyces parvulus DPUA 1573 / Production and characterization of fibrinolytic protease from Streptomyces parvulus DPUA 1573

As proteases fibrinolíticas são capazes de degradar coágulos de fibrina formados dentro dos vasos sanguíneos, evitando a trombose intravascular. Em animais, a tromboflebite, que acomete frequentemente os equinos, ocasiona, em seus casos graves, a obstrução jugular e também um edema de laringe, derivando a obstrução das vias aéreas, o que possibilita um edema cerebral, ocorrendo o óbito do animal. Devido ao fato de o tratamento ser de custo elevado, faz-se necessária a investigação de outras fontesde proteases fibrinolíticas com custos menores e com menos efeitos colaterais. Diante disso, este estudo tem como objetivo produzir e caracterizar proteases fibrinolíticas obtidas de Streptomyces parvulus DPUA 1573. Para produção da enzima, foi utilizado um planejamento fatorial 24 avaliando a concentração da farinha de soja (0,5, 1,0 e 1,5%) e da glicose (0, 0,5 e 1,0g/L), temperatura (28, 32 e 37ºC) e agitação (150, 200 e 250rpm) sobre a biomassa e a atividade fibrinolítica. Pode-se verificar que a protease fibrinolítica apresentou atividade máxima (835U/mL) nas condições de concentração de 1,5% de soja, 1g/L de glicose, 28°C e 150rpm com 48 horas de fermentação. A protease fibrinolítica obtida teve temperatura e pH ótimos de 55°C e pH 9,0, respectivamente. A atividade enzimática foi inibida pelo EDTA, pelo íon Fe2+ e pelo SDS, o que indicou a enzima ser uma metaloprotease. A linhagem Streptomyces parvulus DPUA 1573 foi capaz de produzir protease fibrinolítica, possuindo características bioquímicas favoráveis à aplicação na medicina veterinária e possivelmente humana.(AU)

Fibrinolytic proteases are able to degrade fibrin clot formed in the blood vessel, avoiding intravascular thrombosis. In animals, thrombophlebitis often affects horses, and in severe cases causes obstruction of the jugular and laryngeal edema leading to airway obstruction allowing cerebral edema resulting in the death of the animal. Since treatment is costly, the investigation of other sources of fibrinolytic proteases at lower cost and with fewer side effects is needed. Thus, this study aims to produce and characterize fibrinolytic proteases from Streptomyces parvulus DPUA 1573. For enzyme production, a factorial design was performed to evaluate 24 soybean flour concentration (0.5, 1.0 and 1.5%) and glucose (0, 0.5 and 1.0g/L), temperature (28, 32 and 37°C) and agitation (150, 200 and 250rpm) on biomass and fibrinolytic activity. Fibrinolytic protease showed maximum activity (835 U/mL) under these conditions: 1.5% soybean flour, 1g/L glucose, 28°C, and 150rpm 48 hours of fermentation. The optimal temperature was 55°C and optimal pH was 9.0. Fibrinolytic protease activity was inhibited by EDTA, the ion Fe2+, and by SDS, which indicated that the enzyme is a metallo-protease. The strain Streptomyces parvulus DPUA 1573 was able to produce fibrinolytic protease with biochemical characteristics favorable for application in veterinary and human medicine.(AU)

Coleta de células progenitoras periféricas por aférese automatizada em equino: relato de procedimento / Viability of automated collection of peripheral blood progenitor cells in a horse: report of procedure

The biotechnology used in tendon, bone and joint recoveries in Equine Medicine has improved in recent years. The most used are platelet rich plasma and stem cells from adipose tissue and bone marrow. However, recent studies have shown that stem cells can be found in the bloodstream, also named peripheral blood progenitor cells (CPP). This note aims at reporting the feasibility of automated collection of CPP in horses. The procedure was conducted in an equine, female, Quarter Horses, 2 years old, 385kg. The automated collection of CPP was conducted using apheresis equipment Fresenius- Kabi coupled to C4Y kit. The procedure lasted two hours and 30 minutes without complications, processing 5054mL of whole blood and obtaining 351mL of CPP. Upon completion of the collection, the content of CPP was separated into 10 ml aliquots and immediately stored at -18°C. The automated technique proved to be feasible for horses, but needs improvement in order to achieve greater efficiency and reduce procedure time.(AU)

Produção de biocompostos com atividade antimicrobiana de Streptomyces sp. ante isolados de mastite caprina / Production of biocompounds with antimicrobial activity of Streptomyces sp. Isolated in face of goat mastitis

Actinomicetos são um dos principais produtores de enzimas, vitaminas e metabólitos secundários, destacando-se o gênero Streptomyces, o qual tem uma ampla capacidade de produção de antibióticos eficazes no combate a diferentes microrganismos, entre eles o Staphylococcus sp. Em virtude dessa eficiência no combate a patógenos, o objetivo deste trabalho foi avaliar a produção de metabólitos com atividade antimicrobiana produzidos por 67 Streptomyces isolados de liquens da região amazônica, ante isolados de mastite caprina (Staphylococcus aureus) do estado de Pernambuco, Brasil. Foi utilizado um planejamento fatorial para avaliar a influência das fontes de carbono (glicose) 0%, 0,5% e 1% e de nitrogênio (farinha de soja) 1%, 2,5% e 4% na produção dos antimicrobianos, bem como das variáveis pH, biomassa e atividade antimicrobiana. Dos Streptomyces estudados, o DPUA 1566 foi o que se destacou por formação de halos de inibição entre 18 e 26mm ante os isolados de mastite caprina. Foi possível verificar que a fonte de carbono inibiu a produção de antimicrobianos quando submetidos a uma concentração de glicose de 1%; com a retirada desta, os Streptomyces apresentaram uma elevada capacidade de produção de metabólitos com atividade antimicrobiana tendo potencial para o tratamento de mastite caprina.

Actinomycetes are a leading producer of enzymes, vitamins and secondary metabolites, especially the genus Streptomyces, which have a large capacity for the production of natural antibiotics and are effective against various micro-organisms including Staphylococcus sp. Due to this efficiency in combating micro-organisms, the aim of this study was to evaluate the production of metabolites with antimicrobial activity produced by Streptomyces isolated from lichens 67 in the Amazonia, compared to isolates from goat mastitis (Staphylococcus aureus) in the state of Pernambuco, Brazil. We used a complete factorial design to evaluate the influence of concentrations of carbon sources (glucose) at 0%, 0.5% and 1% and nitrogen (soybean flour) at 1%, 2.5% and 4% in the production of antimicrobial metabolites, and the influence of the pH, microbial biomass and activity variables. Of the studied Streptomyces DPUA 1566 what stood out was the formation of inhibition halos between 18 to 26 mm compared to the isolates from goats mastitis. It was noted that the carbon source inhibited the production of antimicrobial metabolites when subjected to a glucose concentration of 1%. However, after the discontinuation, Streptomyces showed a high capacity to produce metabolites with antimicrobial activity, which has an excellent potential for the treatment of mastitis in goats.

Padronização da técnica de plasmaférese automatizada em equinos / Standardization of automated plasmapheresis in horses

O presente artigo objetivou estudar a viabilidade da técnica de plasmaférese automatizada e padronizá-la em cinco equinos hígidos, bem como apresentar as complicações durante o procedimento, as adequações em relação aos procedimentos em humanos e avaliar a recuperação de volume globular e proteínas plasmáticas totais nos doadores. Os procedimentos foram realizados com o equipamento Fresenius AS104, com duração média de 1h46min, processamento de 5758mL de sangue total e colheita média de 3133mL de plasma. Não foram observadas alterações significativas do volume globular após a plasmaférese automatizada. A recuperação dos níveis plasmáticos de proteínas foi de 91,4% em 96 horas após o procedimento. A plasmaférese automatizada apresentou-se viável para a espécie equina, diminuindo o tempo de recuperação hematimétrica nos doadores.

This paper aimed to study feasibility and standardize the automated plasmapheresis in five healthy horses, showing the complications during the procedure, adjustments in relation to the procedures in humans and assessing the recovery of globular volume and plasma total proteins in donors. The procedures were performed with the Fresenius AS104 equipment, with an average duration of one hour and forty six minutes, processing 5758mL of whole blood and harvest average of 3133mL of plasma. There were no significant variations in globular volume after the automated plasmapheresis. The recovery of plasma total proteins was 91.4% at 96 hours after the procedure. The automated plasmapheresis appeared viable for the equine species, decreasing the time of hematimetric level recovery in donors.

Fontes de fibra na alimentação do pacu: desempenho, composição corporal e morfometria intestinal / Pacu fed different fiber sources: performance, corporal composition and intestinal morphometry

O objetivo deste estudo foi avaliar os efeitos da utilização de diferentes ingredientes fibrosos nas dietas sobre o desempenho, a composição corporal e a morfometria intestinal de juvenis de pacu. Foram avaliadas cinco dietas isoproteicas (23 por cento de proteína digestível), isoenergéticas (3250kcal de energia digestível/kg) e isofibrosas (9 por cento de fibra bruta), sendo a principal fonte fibrosa de cada constituída por farelo de soja, casca de soja, farelo de girassol e polpa cítrica; esta última em dois níveis de inclusão (30 e 45 por cento). Foram utilizados 300 juvenis de pacu (25,12±0,78 gramas), alojados em 25 aquários (200 litros). Os melhores resultados de crescimento e conversão alimentar foram obtidos com as dietas contendo farelo de soja e farelo de girassol. As dietas contendo casca de soja e polpa cítrica prejudicaram o desempenho dos juvenis de pacu, e o efeito negativo foi acentuado com o aumento da inclusão de polpa cítrica. Além disto, somente nos peixes alimentados com a dieta com 45 por cento de polpa cítrica foi observada diminuição na densidade de vilosidades por área no epitélio intestinal. A partir dos resultados obtidos, pode-se concluir que alguns ingredientes fibrosos podem afetar negativamente o desempenho e as características do epitélio intestinal de juvenis de pacu.

The aim of this study was to evaluate the effects of diets containing different fiber sources on the performance, corporal composition and intestinal morphometry of pacu juveniles (Piaractus mesopotamicus). This study evaluated five isoproteic (23 percent digestible protein), isoenergetic (3250kcal digestible energy/kg) and isofibrous (9 percent crude fiber) diets, each one containing a fiber source such as soybean meal, soybean hulls, sunflower meal and citric pulp in two levels of inclusion (30 and 45 percent). A total of 300 pacu juveniles (25,12±0,78g) were stocked in 25 experimental aquariums (200 liters) equipped with aeration and heating system. The best growth and feed conversion results were obtained with diets containing soybean meal and sunflower meal. Diets containing soybean hulls and citrus pulp worsened the juvenile pacu performance. Moreover, only the diet containing 45 percent citrus pulp decreased the villous number per area in the intestinal epithelium. From the results we can conclude that some fibrous ingredient may affect juvenile pacu performance, corporal composition and intestinal epithelium characteristics.

Vascular adaptive responses to physical exercise and to stress are affected differently by nandrolone administration

Androgenic anabolic steroid, physical exercise and stress induce cardiovascular adaptations including increased endothelial function. The present study investigated the effects of these conditions alone and in combination on the vascular responses of male Wistar rats. Exercise was started at 8 weeks of life (60-min swimming sessions 5 days per week for 8 weeks, while carrying a 5 percent body-weight load). One group received nandrolone (5 mg/kg, twice per week for 8 weeks, im). Acute immobilization stress (2 h) was induced immediately before the experimental protocol. Curves for noradrenaline were obtained for thoracic aorta, with and without endothelium from sedentary and trained rats, submitted or not to stress, treated or not with nandrolone. None of the procedures altered the vascular reactivity to noradrenaline in denuded aorta. In intact aorta, stress and exercise produced vascular adaptive responses characterized by endothelium-dependent hyporeactivity to noradrenaline. These conditions in combination did not potentiate the vascular adaptive response. Exercise-induced vascular adaptive response was abolished by nandrolone. In contrast, the aortal reactivity to noradrenaline of sedentary rats and the vascular adaptive response to stress of sedentary and trained rats were not affected by nandrolone. Maximum response for 7-10 rats/group (g): sedentary 3.8 ± 0.2 vs trained 3.0 ± 0.2*; sedentary/stress 2.7 ± 0.2 vs trained/stress 3.1 ± 0.1*; sedentary/nandrolone 3.6 ± 0.1 vs trained/nandrolone 3.8 ± 0.1; sedentary/stress/nandrolone 3.2 ± 0.1 vs trained/stress/nandrolone 2.5 ± 0.1*; *P < 0.05 compared to its respective control. Stress and physical exercise determine similar vascular adaptive response involving distinct mechanisms as indicated by the observation that only the physical exercise-induced adaptive response was abolished by nandrolone.

Opiatergic participation in the thirst-inhibiting effect of acute third ventricle injections of cadmium (Cd 2+ ) and lead (Pb 2+ )

We have previously demonstrated that acute third ventricle injections of both lead and cadmium prevent the dipsogenic response elicited by dehydration or by central injections of dipsogenic agents such as angiotensin II, carbachol and isoproterenol in rats. We have also shown that the antidipsogenic action of cadmium may be due, at least in part, to activation of thirst-inhibitory central serotonergic pathways. In the present paper we show that in Wistar male rats the antidipsogenic effect of both lead acetate (3.0 nmol/rat) and cadmium chloride (3.0 nmol/rat) may be partially dependent on the activation of brain opiatergic pathways since central injections of naloxone (82.5 nmol/rat), a non-selective opioid antagonist, blunt the thirst-inhibiting effect of these metals. One hundred and twenty minutes after the second third ventricle injections, dehydrated animals (14 h overnight) receiving saline + sodium acetate displayed a high water intake (7.90 ñ 0.47 ml/100 g body weight) whereas animals receiving saline + lead acetate drank 3.24 ñ 0.47 ml/100 g body weight. Animals receiving naloxone + lead acetate drank 6.94 ñ 0.60 ml/100 g body weight. Animals receiving saline + saline drank 8.16 ñ 0.66 ml/100 g body weight whilst animals receiving saline + cadmium chloride drank 1.63 ñ 0.37 ml/100 g body weight. Animals receiving naloxone + cadmium chloride drank 8.01 ñ 0.94 ml/100 g body weight. It is suggested that acute third ventricle injections of both lead and cadmium exert their antidipsogenic effect by activating thirst-inhibiting opioid pathways in the brain

Lead (Pb+) and cadmium (Cd2+) inhibit the dipsogenic action of central beta-adrenergic stimulation by isoproterenol

We have previously demonstrated that acute third ventricle injections of both Pb2+ and Cd2+ impair the dipsogenic response elicited by three different situations: dehydration and central cholinergic or angiotensinergic stimulation. beta-Adrenergic activation is part of the multifactorial integrated systems operating in drinking behavior control in the central nervous system. In the present study acute third ventricle injections of Pb2+ (3,30 and 300 pmol/rat) or Cd2+ (0.3, 3 and 30 pmol/rat) blocked the dipsogenic response induced by third ventricle injections of isoproterenol (ISO; 160 nmol/rat) in a dose-dependent manner. Normohydrated animals receiving ISO + NaAc (sodium acetate) or saline (controls) displayed a high water intake after 120 min (ISO + saline = 5.78 ñ O.54 ml/lOO g; ISO + NaAc = 6.00 ñ O.6 ml/lOO g). After the same period,animals receiving ISO but pretreated with PbAc at the highest dose employed (300 pmol/rat) drank O.78 ñ O.23 ml/lOO g while those receiving ISO and pretreated with the highest dose of CdCl(2)(30 pmol/rat) presented a water intake of O.7 ñ O.30 ml/lOO g. Third ventricle injections of CdCl(2) (3 nmol/rat) or PbAc (3 nmol/rat) did not modify food intake in rats deprived of food for 24 h. Thus, general central nervous system depression explaining the antidipsogenic action of the metals can be safely excluded. It is concluded that both Pb2+ and Cd2+ inhibit water intake induced by central beta-adrenergic stimulation.

Calcium channel blockers inhibit the antidipsogenic effect of central injections of zinc in rats

Previous data from our laboratory have indicated that acute third ventricle injections of Zn2+ elicit a significant antidipsogenic response in rats in three different situations: dehydration, and central angiotensinergic or cholinergic stimulation. In the present study we analyzed whether this response depends on voltage-dependent calcium channels. Dehydrated (l4 h of water deprivation, overnight) animals received 2-mul icv injections of zinc acetate (Zn(AC)2; 300 pmol/rat) after pretreatment with the voltage-dependent calcium channel blockers gadolinium (Gd3+;0.03, 3.0 and 30 pmol/rat) or verapamil (VER; 0.027, 0.05 and 0.11 pmol/rat). Both blockers reversed the antidipsogenic effect of third ventricle injections of Zn2+ in a dose-dependent manner. After 120 min, animals pretreated with saline receiving Zn(AC)2 drank 3.10 ñ 0.57 ml/100 g body weight while those pretreated with GD3+ at the highest dose displayed a water intake of 5.45 ñ O.41 ml/l00 g body weight (P<0.01). Animals pretreated with the vehicle of VER receiving Zn(AC)2 drank 3.15 ñ 0.45 ml/l00 g while animals pretreated with VER at the highest dose receiving Zn(AC)2 drank 6.16 ñ 0.62 ml/l00 g (P<0.01). The antidipsogenic effect of Zn(AC)2 seems to be specific since the metal (same dose and injection procedures) did not modify food intake in rats after 24 h of food deprivation. It is suggested that Zn2+ exerts its antidipsogenic effect by activation of mechanism(s) depending on the functional integrity of voltage-dependent calcium channels.

SDZ 216-525, a selective 5-Ht1A receptor antagonist, reverts zinc-induced inhibition of water intake in dehydrate rats

Zinc is found in many brain regions where it participates in important processes such as neurotransmission and neuromodulation. We previously demonstrated that acute third ventricle injection of zinc inhibits wather intake in dehydrated rats. The present study was undertaken to explore a possible link between zinc-induced inhibition of water intake in dehidrated rats and seotonergic systems in the brain. Adult, male Wistar rats had the third ventricle cannulated a week before the experiments. After an overnight period of water deprivation, the animals (N=12 per group) received acute intracerebroventricular injections (2µl) of Zn(Ac)2 (6.7, 67.1 and 67.6 ng/rat). Control animals (N = 12) receives NaAc (671.6 ng/rat). Zinc-treated animals displayed a significant after 120 min was 7.70 ñ 0.50 ml in control (NaAc-treated) dehydrated rats while animals treated with the highest dose of Zn(AC)2 drank 2.63 ñ 0.73 ml. Third ventricle injections of SDZ 216-525, a selective 5-HT1A receptor antagonist, 45 min before zinc administration, generated a dose-dependent reversal of zinc-induced thirst blockade in water-deprived rats. At the highest dose used (10µg/rat), the water intake of the animal after 120 min was 7.30 ñ 0.23 ml, a value equal to that of control animals. These data suggest that zinc may decrease water intake in dehydrated rats by activation of a 5-HT1A receptor-related mechanism