Antibacterial, antibiofilm, and antivirulence potential of the main diterpenes from Copaifera spp. oleoresins against multidrug-resistant bacteria.

To evaluate the antibacterial, antibiofilm and antivirulence potential of the main diterpenes from Copaifera spp. oleoresins against multidrug-resistant (MDR) bacteria. Antimicrobial assays included determination of the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), Minimum Inhibitory Concentration of Biofilm (MICB50), as well as synergistic and antivirulence assays for eight diterpenes against MDR. The tests revealed that two diterpenes (named 1 and 5) showed the best results, with MIC and MBC between 12.5 and 50 µg/mL against most MDR bacteria. These diterpenes exhibited promising MICB50 in concentration between 3.12-25 µg/mL but showed no synergistic antimicrobial activity. In the assessment of antivirulence activity, diterpenes 1 and 5 inhibited only one of the virulence factors evaluated (Dnase) produced by some strains of S. aureus at subinhibitory concentration (6.25 µg/mL). Results obtained indicated that diterpenes isolated from Copaifera oleoresin plays an important part in the search of new antibacterial and antibiofilm agents that can act against MDR bacteria.

Position of the mandibular canal before and after bilateral sagittal split ramus osteotomy: a cone beam computed tomographic study.

The aim of this study was to evaluate the position of the mandibular canal (MC) before and after bilateral sagittal split ramus osteotomy (BSSRO) using cone-beam computed tomography (CT), and to compare the position of the MC in Class II and Class III patients in the preoperative period. Patients were divided into two groups: Class II (n = 38) and Class III (n = 41). Measurements of the superior, inferior, buccal, and lingual distances of the MC in relation to the cortical bone were taken at three levels in the proximal segment of the mandible. Results were analysed using the Kruskal-Wallis test (p < 0.05). In the Class II group the superior distance of the MC at levels 2 and 3, and the inferior distance at level 3 significantly decreased after BSSRO. In the Class III group, no significant differences were found at any level, and the inferior distances at all levels were smaller preoperatively than those in the Class II group. In the Class II group the position of the MC altered in relation to superior and inferior cortical bone after BSSRO. However, the position of the MC remained stable in the Class III group. Our results also suggest a deeper cut in inferior cortical bone in Class III patients.

Antinociception induced by artemisinin nanocapsule in a model of postoperative pain via spinal TLR4 inhibition.

Artemisinin (ART) was initially described for the control of inflammation and pain. However, the mechanisms involved with its antinociceptive effect are still poorly understood. Thus, this present study aimed to investigate the effect of ART in both free and nanocapsulated form on postoperative pain, as well as the participation of the spinal Toll-like receptor 4 (TLR4) in this process. Postoperative pain was induced using the skin/muscle incision retraction (SMIR) model in male Swiss mice. After 3 and 28 days of SMIR, the animals received an intrathecal injection of free or nanocapsulated ART, and the nociceptive threshold was evaluated by von Frey filament test. To evaluate the involvement of the microglia, astrocytes, and TLR4, minocycline (a microglia inhibitor), fluorocitrate (an astrocyte inhibitor), and Lipopolysaccharide Rhodobacter sphaeroides (LPS-RS), a TLR4 antagonist, were intrathecally injected on the third day of SMIR. The levels of spinal TLR4 protein and proinflammatory cytokines tumor necrosis factor-alpha (TNF-α), and interleukin-1-beta (IL-1ß) were quantified by western blot and enzyme-linked immunosorbent assay, respectively. The results showed that free ART reduced postoperative pain (P < 0.001, F5,30 = 7.49, 16.66% for 1000 ng dose; and P < 0.01, F5,30 = 7.49, 14.58% for 500 ng dose) on the 3rd day of SMIR; while the ART nanocapsule had this effect on both the third (P < 0.001; F5,30 = 4.94; 43.75, 39.58 and 72.91% for the 250, 500 and 1000 ng doses, respectively) and 28th (P < 0.05; F5,30 = 7.71; 29.16 and 33.33% for the 500 and 1000 ng doses, respectively) day. The ART nanocapsule had a more potent and longer antinociceptive effect than free ART or morphine. Postoperative pain was also reduced by minocycline and LPS-RS. The ART nanocapsule also reduced the increased levels of TLR4, TNF-α, and IL-1ß induced by SMIR. These data suggest that the ART nanocapsule has a potent analgesic effect on postoperative pain at the spinal level, and this response involves the inhibition of TLR4 and the proinflammatory cytokines TNF-α and IL-1ß.

Energy-dependent effect trial of photobiomodulation on blood pressure in hypertensive rats.

The main purpose of this work was to construct an energy-dependent response curve of photobiomodulation on arterial pressure in hypertension animal model. To reach this objective, we have used a two-kidney one clip (2K-1C) rat model. Animals received acute laser light irradiation (660 nm) on abdominal region using different energy (0.6, 1.8, 3.6, 7.2, 13.8, 28.2, 55.8, and 111.6 J), the direct arterial pressure was measured by femoral cannulation, and systolic arterial pressure (SAP), diastolic arterial pressure (DAP), heart rate (HR), and time of effect were obtained. Our results indicated that 660 nm laser light presents an energy-dependent hypotensive effect, and 28.2 J energy irradiation reached the maximum hypotensive effect, inducing a decreased SAP, DAP, and HR (decrease in SAP: - 19.23 ± 1.82 mmHg, n = 11; DAP: - 9.57 ± 2.23 mmHg, n = 11; HR: - 39.15 ± 5.10 bpm, n = 11; and time of hypotensive effect: 3068.00 ± 719.00 s, n = 11). The higher energy irradiation evaluated (111.6 J) did not induce a hypotensive effect and induced an increase in HR (21.69 ± 7.89 bpm, n = 7). Taken together, our results indicate that red laser energy irradiation from 7.2 to 55.8 J is the effective therapeutic window to reduce SAP, DAP, MAP, and HR and induce a long-lasting hypotensive effect in rats, with effect loss at higher energy irradiation (111.6 J).

Parasitoids diversity in organic Sweet Pepper (Capsicum annuum) associated with Basil (Ocimum basilicum) and Marigold (Tagetes erecta ) / Diversidade de parasitoides em Pimentão Orgânico (Capsicum annuum) associado com Manjericão (Ocimum basilicum ) e Cravo Amarelo (Tagetes erecta)

Abstract The sweet pepper (Capsicum annuum L.) is one of the most important crops in Brazilian farming. Many insect are related to this crop, compromising the quantity and quality of the fruit, representing a production problem. Vegetable diversification is one of the main elements that can be managed for suppressing undesirable insect populations in organic production, once that supports the presence of natural enemies. The basil Ocimum basilicum L. and the marigold Tagetes erecta L. are attractive and nutritious plants for parasitoids, being important candidates for diversified crops. This study evaluated the parasitoids attracted by the association of basil and marigold to organic sweet pepper crop. The experiment comprised three treatments: a) sweet pepper monoculture; b) sweet pepper and basil intercropping; c) sweet pepper and marigold intercropping. Hymenopteran parasitoids were collected over 14 weeks. 268 individuals from 12 families and 41 taxa were collected. Sweet pepper monoculture, sweet pepper-basil intercropping, and sweet pepper-marigold intercropping hosted 40, 98, and 130 individuals and richness of 24, 24, and 23, respectively. Furthermore, the insects of greater abundance in the basil and marigold were different to those collected in the monoculture. The number of parasitoids increased in the associations of sweet pepper with basil and marigold, providing advantages in the use of vegetable diversification for the organic pepper crops management.

Resumo O pimentão (Capsicum annuum L.) é um dos vegetais mais importantes dentre os cultivados no Brasil. Muitos insetos pragas estão relacionados a esta cultura, comprometendo a quantidade e a qualidade do pimentão, tornando-se um grande problema para a produção. A diversificação vegetal é um dos principais componentes a serem manejados para suprimir as populações de pragas na produção orgânica, devido ao apoio a inimigos naturais das pragas. O manjericão ( Ocimum basilicum L.) e cravo amarelo (Tagetes erecta L.) são plantas atrativas e nutritivas para parasitoides, importantes candidatas em cultivos diversificados. Este trabalho avaliou os parasitoides atraídos pela associação de manjericão e cravo amarelo em cultivo de pimentão orgânico. O experimento foi composto por três tratamentos: a) monocultura de pimentão; B) pimentão consorciado com manjericão; C) pimentão consorciado com cravo amarelo. Foram coletadas amostras de parasitoides himenópteros durante 14 semanas. Os espécimes foram identificados até o menor nível de taxonomia possível. Foram coletados 268 indivíduos ao longo de 12 famílias e 41 táxons. Os tratamentos de monocultura, pimentão consorciado com manjericão e pimentão consorciado com cravo amarelo resultaram em 40, 98 e 130 indivíduos e riqueza de espécies de 24, 24 e 23, respectivamente. Além disso, os insetos de maior abundância coletados no manjericão e cravo amarelo, foram diferentes daqueles coletados na monocultura. Houve um aumento na abundância de parasitoides nas associações de pimentão com manjericão e cravo amarelo, proporcionando vantagens no uso da diversificação vegetal, podendo ser usada no manejo do pimentão orgânico.

Ocorrência de Campylobacter e Enterobacteriaceae em aves silvestres e frangos de corte / Campylobacter and Enterobacteriaceae in wild birds and poultry

As aves silvestres podem ser reservatório de bactérias patogênicas e atuar como veiculadoras desses microrganismos para o ambiente, os animais domésticos e o homem. Portanto, o objetivo deste trabalho foi verificar a ocorrência de Campylobacter spp., Yersinia enterocolitica e Salmonella enterica em aves silvestres capturadas nas áreas próximas de aviários e em frangos de corte alojados nesses estabelecimentos, além de verificar a presença dos genes cdtA, cdtB e cdtC nos isolados de Campylobacter e identificar os sorotipos de Salmonella encontrados. Amostras de fezes de 189 aves silvestres capturadas com redes de neblina nas áreas próximas de 10 aviários e de 200 frangos de corte foram processadas para pesquisa de Campylobacter spp., S. enterica e Y. enterocolitica. Duas espécies de aves silvestres, Sicalis flaveola (canário-da-terra) e Zonotrichia capensis (tico-tico), foram positivas para Salmonella e Campylobacter, respectivamente. Foram isolados Campylobacter spp., S. enterica e Y. enterocolitica de frangos. Todos os isolados de Campylobacter analisados apresentaram os genes cdt. Em dois aviários, Campylobacter foi isolado tanto de frangos como de aves silvestres, entretanto a contaminação mútua entre essas aves não foi comprovada. Este foi o primeiro relato de isolamento de Campylobacter de Z. capensis e de Salmonella do sorotipo Derby de S. flaveola.(AU)

Wild birds can be reservoirs of pathogenic bacteria and act as carriers of these microorganisms to the environment, domestic animals, and humans. Therefore, this study had as objective to verify the occurrence of Campylobacter spp., Yersinia enterocolitica and Salmonella enterica in wild birds captured in the surroundings of the aviaries and in the broilers housed in these establishments. The presence of the cdtA, cdtB and cdtC genes in Campylobacter isolates was also investigated and Salmonella serotypes were identified. Stool samples from 189 wild birds captured with mist nets in around 10 aviaries and from 200 broilers were processed for Campylobacter spp., S. enterica and Y. enterocolitica research. Two species of wild birds, Sicalis flaveola (Saffron Finch) and Zonotrichia capensis (Rufous-collared Sparrow) were positive for Salmonella and Campylobacter, respectively. Campylobacter spp., S. enterica and Y. enterocolitica were isolated from broilers. The cdt genes were found in all Campylobacter isolates. In two aviaries, Campylobacter was isolated from both broilers and wild birds, however the mutual contamination among these birds has not been shown. This was the first report of Campylobacter isolation from Z. capensis and of Derby Salmonella serotype isolation from S. flaveola.(AU)

Avaliação morfofuncional do enxerto de túnica albugínea suína na cistoplastia em ratos / Morpho-fuctional evaluation of porcine tunica albuginea as a graft for cystoplasty in rats

O objetivo deste trabalho é avaliar o uso da túnica albugínea suína na cistoplastia em ratos, avaliando funcionalidade, capacidade de reparação do órgão e possibilidades de complicações. Foram selecionados 30 ratos Wistar, machos, de seis meses de idade, divididos em: um grupo teste (TA), em que os animais receberam o enxerto de túnica albugínea suína após a cistectomia parcial e um grupo controle (C), em que os animais sofreram somente a cistectomia parcial. Os animais pertencentes a ambos os grupos foram divididos igualmente em subgrupos de cinco animais cada, que sofreram eutanásia em sete, 28 e 42 dias de pós-operatório. Foi realizada uma análise macroscópica e, posteriormente, uma análise histopatológica da região da ferida cirúrgica. Aos sete e 28 dias, os animais pertencentes ao grupo C e ao grupo TA apresentaram urotelização, regeneração da lâmina própria e da musculatura, porém o grupo TA apresentou menores sinais inflamatórios e maior organização tecidual, principalmente com relação à formação das fibras musculares. Aos 42 dias de pós-operatório, ambos os grupos já apresentavam características histológicas normais. Concluiu-se que o enxerto de túnica albugínea suína obteve sucesso na regeneração da bexiga de ratos, mantendo a funcionalidade do órgão, sem rejeição, e favorecendo a migração celular.(AU)

The aim of this study is to evaluate porcine tunica albuginea as a graft for cystoplasty in rats, regarding bladder function, capacity and possible complications. 30 male Wistar rats with six monthes of age have been selected and separated into two different groups: A test group (TA) in which the animals received a tunica albuginea graft after partial cystectomy and a control group (C) in which partial cystectomy was performed, followed by bladder suture. In each group the animals were euthanized at seven, 28 and 42 days after surgery. Macroscopic and Histological analysis have been performed. At seven and 28 days after surgery the samples from both groups had urothelial lining upon a lamina propria and smooth muscle fibers in regeneration process. However, the TA group showed less inflammatory signs and more organized structure, mainly regarding the smooth muscle formation. At 42 days after surgery all groups showed a bladder wall structure qualitatively identical to the normal tissue. We could conclude that tunica albuginea graft is able to maintain bladder function and support cellular migration without any kind of rejection.(AU)

Hypotensive acute effect of photobiomodulation therapy on hypertensive rats.

The purpose of this study was to evaluate the acute effect of photobiomodulation therapy (PBM) on arterial pressure in hypertensive and normotensive rats with application in an abdominal region. Normotensive (2K) and hypertensive (2K-1C) wistar rats were treated with PBM. Systolic arterial pressure (SAP), diastolic arterial pressure (DAP), mean arterial pressure (MAP) and heart rate (HR) were measured before, during and after PBM application. The nitric oxide (NO) serum concentration was measured before and after PBM application. Vascular reactivity study was performed in isolated thoracic aortas. Aluminum gallium arsenide (GaAlAs) diode laser was used, at 660nm wavelength and 100mW optical output. The PBM application induced a decrease of SAP in 2K-1C rats. In 2K rats, the PBM application had no effect on SAP, DAP and MAP. Moreover, the magnitude of hypotensive effect was higher in 2K-1C than in 2K rats. The PBM application induced a decrease of HR in 2K-1C and 2K, with higher effect in 2K-1C rats. In 2K-1C, the hypotensive effect induced by PBM was longer than that obtained in 2K rats. PBM application induced an elevation of NO concentration in serum from 2K-1C and 2K rats, with higher effect in 2K-1C. In isolated aortic rings PBM effect is dependent of NO release, and is not dependent of nitric oxide synthase (NOS) activation. Our results indicate that the abdominal acute application of PBM at 660nm is able to induce a long lasting hypotensive effect in hypertensive rats and vasodilation by a NO dependent mechanism.

Parasitoids diversity in organic Sweet Pepper (Capsicum annuum) associated with Basil (Ocimum basilicum) and Marigold (Tagetes erecta )

Abstract The sweet pepper (Capsicum annuum L.) is one of the most important crops in Brazilian farming. Many insect are related to this crop, compromising the quantity and quality of the fruit, representing a production problem. Vegetable diversification is one of the main elements that can be managed for suppressing undesirable insect populations in organic production, once that supports the presence of natural enemies. The basil Ocimum basilicum L. and the marigold Tagetes erecta L. are attractive and nutritious plants for parasitoids, being important candidates for diversified crops. This study evaluated the parasitoids attracted by the association of basil and marigold to organic sweet pepper crop. The experiment comprised three treatments: a) sweet pepper monoculture; b) sweet pepper and basil intercropping; c) sweet pepper and marigold intercropping. Hymenopteran parasitoids were collected over 14 weeks. 268 individuals from 12 families and 41 taxa were collected. Sweet pepper monoculture, sweet pepper-basil intercropping, and sweet pepper-marigold intercropping hosted 40, 98, and 130 individuals and richness of 24, 24, and 23, respectively. Furthermore, the insects of greater abundance in the basil and marigold were different to those collected in the monoculture. The number of parasitoids increased in the associations of sweet pepper with basil and marigold, providing advantages in the use of vegetable diversification for the organic pepper crops management.

Resumo O pimentão (Capsicum annuum L.) é um dos vegetais mais importantes dentre os cultivados no Brasil. Muitos insetos pragas estão relacionados a esta cultura, comprometendo a quantidade e a qualidade do pimentão, tornando-se um grande problema para a produção. A diversificação vegetal é um dos principais componentes a serem manejados para suprimir as populações de pragas na produção orgânica, devido ao apoio a inimigos naturais das pragas. O manjericão ( Ocimum basilicum L.) e cravo amarelo (Tagetes erecta L.) são plantas atrativas e nutritivas para parasitoides, importantes candidatas em cultivos diversificados. Este trabalho avaliou os parasitoides atraídos pela associação de manjericão e cravo amarelo em cultivo de pimentão orgânico. O experimento foi composto por três tratamentos: a) monocultura de pimentão; B) pimentão consorciado com manjericão; C) pimentão consorciado com cravo amarelo. Foram coletadas amostras de parasitoides himenópteros durante 14 semanas. Os espécimes foram identificados até o menor nível de taxonomia possível. Foram coletados 268 indivíduos ao longo de 12 famílias e 41 táxons. Os tratamentos de monocultura, pimentão consorciado com manjericão e pimentão consorciado com cravo amarelo resultaram em 40, 98 e 130 indivíduos e riqueza de espécies de 24, 24 e 23, respectivamente. Além disso, os insetos de maior abundância coletados no manjericão e cravo amarelo, foram diferentes daqueles coletados na monocultura. Houve um aumento na abundância de parasitoides nas associações de pimentão com manjericão e cravo amarelo, proporcionando vantagens no uso da diversificação vegetal, podendo ser usada no manejo do pimentão orgânico.

Agonistic 4-1BB antibody fails to reduce disease burden during acute respiratory syncytial virus (RSV) infection.

Respiratory Syncytial Virus (RSV) remains a leading cause of infant morbidity and mortality worldwide. Despite this, there are limited therapeutic options. CD8 T cells have an integral role in controlling viral infections; strategies to enhance these responses may be clinically relevant. The T cell costimulatory receptor, 4-1BB, is known to play a role in expansion of antiviral CD8 T cells. In this study, we examined the effect of agonistic 4-1BB antibody at the time of RSV infection in mice. We show that this antibody did not improve outcomes in the setting of RSV infection but rather, led to increased weight loss and a reduction in RSV specific CD8 T cells in the lung. This work suggests caution in the use of agonistic 4-1BB antibody in the setting of viral infections.

Gastric Ulcers in Middle-Aged Rats: The Healing Effect of Essential Oil from Citrus aurantium L. (Rutaceae).

The elderly population has experienced increased life expectancy as well as the increased incidence of gastric ulcers. The peels of fruits from Citrus aurantium L., popularly known in Brazil as orange bitter, are commonly used asatea form for the treatment of gastrointestinal tract disorders, such as ulcer and gastritis. We evaluated the healing effects of essential oil from the peels of Citrus aurantium fruits (OEC) on gastric ulcers in middle-aged rats. We examined the effects of a 14-day chronic OEC treatment on gastric mucosa in middle-aged male Wistar rats that were given acetic-acid-induced gastric lesions by morphometric and immunohistological analyses. Oral OEC treatment significantly reduced the lesion area (76%) within the gastric mucosa and significantly increased (P < .05) the height of regenerated mucosa (59%) when compared to the negative control group. Immunohistochemical analysis of the molecular markers such as COX-2, HSP-70, VEGF, and PCNA in the gastric mucosa confirmed that OEC treatment induced healing effects by increasing the number of new blood vessels and by augmenting gastric mucus in the mucosa glands. These results suggest that the oil from Citrus aurantium effectively heals gastric ulcers in middle-aged animals; however, safe use of OEC demands special care and precautions.

Avaliação de diferentes vias vacinais para vacinação contra o vírus da doença de Newcastle em aves de fundo de quintal / Assessment of different vaccination approaches against Newcastle disease virus in domestic backyard poultry

Foram avaliadas três vias de aplicação vacinal contra o vírus da doença de Newcastle em aves de criatório de fundo de quintal (AFQ) jovens e adultas. Um total de 135 AFQ foram distribuídas em tratamentos distintos de acordo com a via vacinal: via ocular (VO), água de bebida (VAB) e alimentar (VA). Cada tratamento foi representado por 40 aves (20 jovens e 20 adultas) e utilizou-se um grupo-controle de 15 aves não vacinadas. O programa de vacinação estabelecido constou de uma primovacinação e dois reforços vacinais, utilizando-se a cepa La Sota. Para aves jovens, os títulos obtidos pelas VO e VAB não diferiram aos 15, 45 e 140 dias, mas houve diferenças nos títulos das aves vacinadas pela VA. Nas aves adultas, a vacinação pela VO apresentou resultados mais elevados que as vacinações pelas VAB e VA na primeira resposta, aos 15 dias. Aos 45 dias, os títulos obtidos pela VAB foram mais baixos que os obtidos pela VO, e, aos 140 dias, não houve diferença entre as três vias avaliadas. Concluiu-se que as vacinações pelas VO e VAB constituem alternativas eficazes para vacinação de AFQ jovens e adultas.

Three ways of vaccination against Newcastle Disease Virus (NDV) were evaluated in young and adults domestic backyard poultry (DBP). A total of 135 DBP was submitted to three different administration routes of ND vaccine: eye-drop, drinking water, and feed. Each treatment consisted of 40 birds (20 young and 20 adult) and a control group of 15 unvaccinated birds. The treatment consisted of a first vaccination and two boosters, using La Sota strain. For young birds, the eye-drop and drinking water vaccinations presented no differences at 15, 45, and 140 days, differing from the titers obtained by birds treated by feed vaccination method. In the adult birds, the eye-drop administration presented higher titers than by drinking water and feed approaches in the first response to the vaccination at 15 days. At 45 days, the results obtained by the drinking water had lower titers than those from the eye-drop. The three vaccination methods presented no difference at 140 days. In conclusion, the vaccination by eye-drop and drinking water methods constituted an efficient alternative of vaccination for adult and young DBP against Newcastle virus.

Efeito da adição de trolox e pentoxifilina na motilidade, integridade do acrossoma e do DNA de espermatozoides equinos após descongelação / Effect of trolox and pentoxifylline on motility and integrity of, acrossome and DNA of equine spermatozoa after thawing

Três garanhões foram utilizados para estudar o efeito da adição de trolox e pentoxifilina na motilidade, integridade do acrossoma e DNA de espermatozoides pós-descongelação. Para congelação, utilizou-se Tris-gema com glicerol (5 por cento) em máquina de congelação de sêmen. As amostras foram descongeladas a 37ºC durante 30 segundos e tratadas com: T1= 150µL de sêmen + 150µL de Tris; T2= 150µL de sêmen + 150µL de Tris + 120µM/mL de trolox; T3= 150µL de sêmen + 150µL de Tris + 3,5mM de pentoxifilina e T4= 150µL de sêmen + 150µL de Tris + 3,5mM de pentoxifilina + 120µM/mL de trolox. Após 0, 60 e 120 minutos de incubação (37ºC), as amostras foram analisadas quanto à motilidade, vigor, integridade de acrossoma e DNA. Não houve diferença (P>0,05) entre tratamentos após 0 e 60 minutos de incubação em todos os parâmetros estudados. Após 120 minutos de incubação, verificou-se maior porcentual (P<0,05) de células com motilidade total e progressiva nas amostras do T2. Conclui-se que a adição de trolox após descongelação do sêmen equino preserva a motilidade total e progressiva dos espermatozoides submetidos à incubação a 37ºC durante 120 minutos.

Three stallions were used to study the effect of trolox and pentoxifylline addition on the motility and integrity of acrossome and DNA equine spermatozoa after thawing. Tris-egg-yolg diluent with glycerol (5 percent) were used to freeze the semen samples in a freezing machine. The samples were thawed at 37ºC during 30 seconds and treated with: T1=150µL of semen + 150µL of Tris; T2= 150µL of semen + 150µL of Tris +150mM/mL of trolox; T3= 150µL of semen + 150µL Tris +3.5mM of pentoxifylline; and T4= 150µL of semen + 150µL of Tris + 3.5mM of pentoxifylline + 150mM of trolox. After 0, 60, and 120 minutes of incubation (37ºC), the samples were analyzed to motility, vigor, and integrity of acrossome and DNA. There was no difference (P>0.05) among treatments considering 0 and 60 minutes of incubation in all studied parameters. After 120 minutes of incubation, it was observed higher percentage (P<0.05) of cells with total and progressive motility in the samples of T2. It can be concluded that the trolox addition after thawing of equine semen preserved total and progressive motility of the sperm incubated at 37ºC during 120 minutes.

Efeito do uso de pentoxifilina no período neonatal sobre a produção espermática em ratos Wistar adultos / Effect of pentoxifylline during neonatal period on spermatic production in adult Wistar rats

Utilizaram-se doses crescentes de pentoxifilina em ratos Wistar neonatos visando aumentar a produção espermática em animais adultos. Trinta e sete animais foram distribuídos de acordo com os tratamentos: não tratados (n=10) e tratados com 1mg/kg (n=10), 5mg/kg (n=9) e 10mg/kg (n=8) de pentoxifilina (IP). Aos 90 dias, os animais foram anestesiados e perfundidos intracardiacamente com solução fixadora. Os testículos foram processados rotineiramente para inclusão em resina plástica à base de glicol metacrilato. Cortes histológicos de 4µm de espessura foram corados em azul de toluidina/borato de sódio a 1 por cento e analisados histometricamente. O número de células de Sertoli por secção transversal diminuiu nos grupos tratados com 5mg/kg e 10mg/kg em relação aos grupos controle e tratado com 1mg/kg. O índice de células de Sertoli aumentou nos animais tratados com 5mg/kg em comparação aos do grupo-controle. A utilização da pentoxifilina não foi capaz de induzir aumento na população das células de Sertoli e produção espermática em ratos adultos.

Increasing doses of pentoxifylline were administrated to newborn Wistar rats in order to augment Sertoli cell number and sperm production in the adult rats. Thirty-seven neonate Wistar rats were distributed in four groups: control (n=10) and treated with 1mg/kg (n=10), 5mg/kg (n=9), and 10mg/kg (n=8) of pentoxifylline. At 90 days, the animals were submitted to anesthesia and intracardiac perfusion. Testes were colleted and routinely processed for inclusion in plastic resin with glycol methacrylate. Histological sections (4µm) were stained in toluidine blue/sodium borate (1 percent) and analyzed. Number of Sertoli cell per transversal section of seminiferous tubule had significant reduction in the groups treated with 5mg/kg and 10mg/kg of pentoxifylline as compared to control and the group that received 1mg/kg (P<0.05). The Sertoli cell index significantly increased in the group treated with 5mg/kg compared to control group. Pentoxifylline did not cause increase in the number of Sertoli cells and daily sperm production in adult male rats.

Estado antioxidante do sangue como indicador da eficiência do treinamento em nadadores / Blood antioxidant status: a parameter to stablish the swimmer perfomance

A prática regular de exercícios deve levar à regulaçäo da atividade das enzimas antioxidante do músculo esquelético e os oxidantes atuariam como indutores de tal efeito. O principal objetivo deste estudo foi determinar respostas metabólicas adaptativas ao estresse oxidativo em nadadores antes e após um período de treinamento. O estudo foi realizado com doze atletas da equipe de nataçäo do Yara Clube de Marília, submetidos a um período de trinta dias de treinamento, seis vezes por semana. Os resultados demonstraram que exercícios de intensidade variável foram associados com reduçäo na capacidade antioxidante do plasma e respostas adaptativas no sistema da GSH juntamente com a regulaçäo positiva da atividade da SOD eritrocítica. Portanto, concluímos que o exercício físico prolongado em atletasmoderadamente treinados, desenvolve uma resposta do sistema antioxidante numa tentativa de suplantar a geraçäo de radicais livres advindos do metabolismo oxidativo e que o estado antioxidante intraeritrocítico e do plasma säo eficientes indicadores do estresse oxidativo.

Noncovalent interaction between ubiquitin and the human DNA repair protein Mms2 is required for Ubc13-mediated polyubiquitination.

Ubiquitin-conjugating enzyme variants share significant sequence similarity with typical E2 (ubiquitin-conjugating) enzymes of the protein ubiquitination pathway but lack their characteristic active site cysteine residue. The MMS2 gene of Saccharomyces cerevisiae encodes one such ubiquitin-conjugating enzyme variant that is involved in the error-free DNA postreplicative repair pathway through its association with Ubc13, an E2. The Mms2-Ubc13 heterodimer is capable of linking ubiquitin molecules to one another through an isopeptide bond between the C terminus and Lys-63. Using highly purified components, we show here that the human forms of Mms2 and Ubc13 associate into a heterodimer that is stable over a range of conditions. The ubiquitin-thiol ester form of the heterodimer can be produced by the direct activation of its Ubc13 subunit with E1 (ubiquitin-activating enzyme) or by the association of Mms2 with the Ubc13-ubiquitin thiol ester. The activated heterodimer is capable of transferring its covalently bound ubiquitin to Lys-63 of an untethered ubiquitin molecule, resulting in diubiquitin as the predominant species. In (1)H (15)N HSQC ((1)H (15)N heteronuclear single quantum coherence) NMR experiments, we have mapped the surface determinants of tethered and untethered ubiquitin that interact with Mms2 and Ubc13 in both their monomeric and dimeric forms. These results have identified a surface of untethered ubiquitin that interacts with Mms2 in the monomeric and heterodimeric form. Furthermore, the C-terminal tail of ubiquitin does not participate in this interaction. These results suggest that the role of Mms2 is to correctly orient either a target-bound or untethered ubiquitin molecule such that its Lys-63 is placed proximally to the C terminus of the ubiquitin molecule that is linked to the active site of Ubc13.

Crystal structure of the human ubiquitin conjugating enzyme complex, hMms2-hUbc13.

The ubiquitin conjugating enzyme complex Mms2-Ubc13 plays a key role in post-replicative DNA repair in yeast and the NF-kappaB signal transduction pathway in humans. This complex assembles novel polyubiquitin chains onto yet uncharacterized protein targets. Here we report the crystal structure of a complex between hMms2 (Uev1) and hUbc13 at 1.85 A resolution and a structure of free hMms2 at 1.9 A resolution. These structures reveal that the hMms2 monomer undergoes a localized conformational change upon interaction with hUbc13. The nature of the interface provides a physical basis for the preference of Mms2 for Ubc13 as a partner over a variety of other structurally similar ubiquitin-conjugating enzymes. The structure of the hMms2-hUbc13 complex provides the conceptual foundation for understanding the mechanism of Lys 63 multiubiquitin chain assembly and for its interactions with the RING finger proteins Rad5 and Traf6.

Efeito de níveis de vitamina E na dieta sobre o desempenho e concentraçäo de alfa-tocoferol na carne de frangos de corte / Effect of dietary vitamin E on performance and alpha-tocoferol concentration in meat of broiler

Este experimento foi realizado com o objetivo de avaliar o efeito da suplementaçäo de diferentes níveis de vitamina E (NVE) na dieta sobre o desempenho de frangos de corte de um a 42 dias de idade, e sobre a concentraçäo de alfa-tocoferol (AT) na carne de peito das aves aos 45 dias de idade. Foram alojados 480 pintos de um dia, da linhagem Ross, em 16 boxes de 3,00 x 2,20m, com 30 aves de ambos os sexos em cada boxe. A raçäo e a água foram fornecidas ad libitum. As aves receberam uma dieta inicial (1 a 21 dias), com 21,2 por cento de PB e 2934kcal e EM/kg, e uma dieta de crescimento (22 a 42 dias), com 18 por cento de PB e 3035kcal de EM/kg. Ambas as dietas foram suplementadas com quatro NVE: 25, 250, 500 e 750mg/kg. O delineamento experimental foi o inteiramento ao acaso, constituído de quatro tratamentos (NVE), com quatro repetiçöes, cada uma com 30 aves. Verificou-se que o peso corporal, o ganho de peso e a conversço alimentar aos 42 dias de idade foram significativamente influenciados (P<0,05) pelos NVE utilizados na dieta. Houve aumento linear para peso e ganho de peso, e melhoria linear da conversäo alimentar à medida que se elevou o nível da suplementaçäo de vitamina E (VE) da dieta. O consumo de raçäo e a viabilidade näo foram influenciados (P>0,05) pelos NVE. Observou-se efeito quadrático (P<0,05) do NVE na dieta sobre a deposiçäo de AT na carne de peito das aves, sem diferença entre sexos. Máxima concentraçäo de AT foi verificada na carne de peito de aves que foram alimentadas com dieta de 500mg de VE/kg. Desse modo, conclui-se que para cada aumento de 25mg de VE/kg na dieta, espera-se um aumento de 5,49g no peso corporal, melhoria de 0,0038 pontos na conversäo alimentar, e uma aumento de 8,57ug de AT/g de carne de peito de frangos de corte

Efeito da suplementaçäo da vitamina E sobre a concentraçäo de alfa-tocoferol na gema e nos tecidos de balanço de nitrogênio em reprodutora pesada / Effect of vitamin E on yolk and tissues alfa-tocopherol concentration and nitrogen balance in broiler breeder hen

Avaliou-se efeito da suplementaçäo da dieta com diferentes níveis de vitamina E (25, 50, 75 e 100mg/kg) sobre a concentraçäo de alfa-tocoferol (AT) na gema, no fígado, no soro e na excreta, e sobre a retençäo de AT e de nitrogênio (N) em reprodutoras pesadas. Dezesseis aves da linhagem Ross, com 35 semanas de idade, foram alojadas em gaiolas de recria, equipadas com grade coletora de ovos, bebedouros e comedouros individuais e com bandeja metálica individual. Cada tratamento foi constituído de quatro repetiçöes. A dieta experimental foi fornecida durante 25 dias, sendo 15 dias para adaptaçäo das aves à dieta experimental e 10 dias para a coleta de ovos e de excreta para análise de AT e N. Verificou-se aumento linear (P<0,05) no consumo, na excreçäo e na retençäo de AT nas aves em funçäo do aumento do nível de vitamina E na dieta, refletindo um aumento linear (P<0,05) na concentraçäo de AT na gema, no fígado, no soro e na excreta. Observou-se que o consumo, a excreçäo, a retençäo e a concentraçäo de N (balanço de N) no ovo näo foram influenciados (P>0,05) pelos níveis de vitamina E adicionados à dieta