Resumo O objetivo deste trabalho foi determinar o tamanho ótimo de parcela e o número de repetições para avaliação da massa de matéria fresca de azevém semeado em filas. Setenta ensaios de uniformidade foram realizados com azevém 'Barjumbo', em 16 unidades experimentais básicas (UEBs) de 0, 51 m 2 cada uma. A massa de matéria fresca do azevém nas UEBs de 18, 18, 6, 6 e 22 ensaios de uniformidade foi determinada, respectivamente, aos 130, 131, 133, 134 e 137 dias após a semeadura. O tamanho ótimo de parcela foi determinado pelo método da máxima curvatura do coeficiente de variação. O número de repetições foi determinado em cenários formados por combinações de número de tratamentos e de diferenças entre médias a serem detectadas como significativas pelo teste de Tukey, a 5% de probabilidade. O tamanho ótimo de parcela oscilou de 1,73 a 3,18 m2, e o coeficiente de variação no tamanho ótimo de parcela de 7,58 a 13,96%. O número de repetições oscilou de 3,95 (~4) a 32,27 (~33), conforme o delineamento, o número de tratamentos e a diferença mínima adotada. O tamanho ótimo de parcela é de 2,29 m2 e, em experimentos com até 50 tratamentos, são necessárias oito repetições para identificar como significativas as diferenças entre médias de tratamentos de 20,24%.

Abstract The objective of this work was to determine the optimal plot size and the number of replicates for the evaluation of the fresh weight of ryegrass sowed in rows. Seventy uniformity trials were performed with 'Barjumbo' ryegrass, in 16 basic experimental units (BEUs) of 0.51 m2 each. The fresh weight of ryegrass in the BEUs of 18, 18, 6, 6, and 22 uniformity trials was determined, respectively, at 130, 131, 133, 134, and 137 days after sowing. The optimal plot size was determined through the method of the maximum curvature of the coefficient of variation. The number of replicates was determined in scenarios formed by combinations of treatments and differences between means to be detected as significant by Tukey’s test, at 5% probabilit y. The optimal plot size ranged from 1.73 to 3.18 m2, and the variation coefficient in the optimal plot size from 7.58 to 13.96%. The number of replicates varied from 3.95 (~4) to 32.27 (~33), depending on the experimental design, the number of treatments, and the adopted minimum difference. The optimal plot size is 2.29 m2, and, in experiments with up to 50 treatments, eight replicates are required to identify as significant the differences between treatment means of 20.24%.

Resumen Antecedentes: La tinción con azul cresil brillante (BCB) es un método no invasivo para seleccionar ovocitos aptos para el desarrollo embrionario. Por tanto, es una herramienta útil para selecionar los ovocitos de mejor calidad en temporadas de restricción de forraje. Objetivo: Evaluar el efecto de la estacionalidad sobre la maduración nuclear y calidad de los ovocitos seleccionados por el test BCB. Métodos: Los complejos cumulus-oophorus (CCOs) fueron obtenidos durante el verano y el invierno de 2010 y 2011. Los CCOs seleccionados se mantuvieron durante 90 min a 38,5 oC en una incubadora de CO2 en un medio TCM 199 con 10% de suero fetal bovino y antibióticos, suplementado con 26 µM de azul cresil brillante. Luego se separaron según el color del citoplasma (BCB+ -azul y BCB− -incoloro). Posteriormente, los CCOs se maduraron durante 22 h. La evaluación de la maduración nuclear se realizó a las 22 h de cultivo. Resultados: La proporción de ovocitos BCB− fue mayor en el invierno de 2010, pero no hubo un aumento de ese grupo en el invierno de 2011. El porcentaje de ovocitos que alcanzaron la etapa de metafase II fue mayor en el grupo control y BCB+ con respecto al grupo BCB−. Conclusión: La estación del año influye en el porcentaje de ovocitos más aptos para la producción de embriones en situaciones donde las donadoras de ovocitos reciben alimentación a base de pastos, ya que este método fue eficaz para determinar el efecto de la estacionalidad en la competencia de ovocitos bovinos en alcanzar la maduración nuclear.

Abstract Background: The brilliant cresyl blue (BCB) staining is a non-invasive test to select the best-suited oocytes for embryonic development. This makes it a useful tool to select best-quality oocytes at the times of the year when there is forage restriction. Objective: To evaluate the effect of seasonality on the nuclear maturation and quality of oocytes selected by the BCB test. Methods: The cumulus-oophorus complexes (COCs) were obtained in summer and winter of 2010 and 2011. Selected COCs were maintained for 90 min at 38.5 °C in a CO2 incubator, in TCM 199 medium containing 10% fetal bovine serum and antibiotics, and supplemented with 26 µM brilliant cresyl blue. Afterwards, they were divided according to the ooplasm staining (BCB+ -blue; BCB− -unstained). Subsequently, COCs were matured for 22 h. Nuclear maturation was evaluated at 22 h of culture. Results: The proportion of BCB− oocytes was higher in the winter of 2010, but there was no increase in this group in the winter of 2011. The percentage of oocytes that reached metaphase II was higher in control and BCB+ groups in relation to oocytes from BCB− group. Conclusion: The season of the year influences the percentage of oocytes best suited for embryonic production in situations in which oocyte donors receive pasture-based feeding, since the method was effective in determining the effect of seasonality on the competence of bovine oocytes to reach nuclear maturation.

Resumo Antecedentes: O método do azul cresil brilhante (BCB) não é invasivo e seleciona ovócitos mais aptos ao desenvolvimento embrionário. Portanto é ferramenta útil para selecionar ovócitos de melhor qualidade em épocas do ano que ocorre restrição de pastagem. Objetivo: Avaliar o efeito da sazonalidade sobre a maturação nuclear e a qualidade dos ovócitos selecionados pelo teste BCB. Métodos: Os complexos cumulus oophorus (CCOs) foram obtidos no verão e inverno de 2010 e 2011. Os CCOs selecionados foram mantidos por 90 min, a 38,5 °C, em incubadora de CO2, em meio TCM 199 contendo 10% de soro fetal bovino e antibióticos, e suplementado com 26 µM de azul cresil brilhante. Em seguida, estes foram divididos de acordo com a coloração do citoplasma (BCB+ -azuis e BCB− -incolores). Então os CCOs foram maturados durante 22 h. A avaliação da maturação nuclear foi realizada às 22 h de cultivo. Resultados: A proporção dos ovócitos BCB− foi maior no inverno de 2010, mas não houve aumento desse grupo no inverno de 2011. O percentual de ovócitos que atingiu o estágio de metáfase II foi maior no controle e no grupo BCB+ em relação ao grupo BCB−. Conclusão: A estação do ano influencia o percentual de ovócitos mais aptos a produção de embriões, em situações onde as doadoras de ovócitos recebem alimentação baseada em pastagens, já que o método se mostrou efetivo para determinação do efeito da sazonalidade sobre a competência de ovócitos bovinos em atingirem a maturação nuclear.

Localised cutaneous leishmaniasis (LCL) is the most common form of cutaneous leishmaniasis characterised by single or multiple painless chronic ulcers, which commonly presents with secondary bacterial infection. Previous culture-based studies have found staphylococci, streptococci, and opportunistic pathogenic bacteria in LCL lesions, but there have been no comparisons to normal skin. In addition, this approach has strong bias for determining bacterial composition. The present study tested the hypothesis that bacterial communities in LCL lesions differ from those found on healthy skin (HS). Using a high throughput amplicon sequencing approach, which allows for better populational evaluation due to greater depth coverage and the Quantitative Insights Into Microbial Ecology pipeline, we compared the microbiological signature of LCL lesions with that of contralateral HS from the same individuals.Streptococcus, Staphylococcus,Fusobacterium and other strict or facultative anaerobic bacteria composed the LCL microbiome. Aerobic and facultative anaerobic bacteria found in HS, including environmental bacteria, were significantly decreased in LCL lesions (p < 0.01). This paper presents the first comprehensive microbiome identification from LCL lesions with next generation sequence methodology and shows a marked reduction of bacterial diversity in the lesions.

JUSTIFICATIVA E OBJETIVOS: As citocinas pró-inflamatórias têm função importante na fisiopatologia das síndromes dolorosas neuropáticas. O objetivo desse estudo foi avaliar os níveis plasmáticos de citocinas pró-inflamatórias antes e após o tratamento com tramadol em pacientes com hérnia discal e síndrome do túnel do carpo e compará-los com indivíduos normais. MÉTODO: Investigou-se 38 pacientes com dor neuropática por hérnia discal ou síndrome do túnel do carpo. Todos os pacientes foram tratados com tramadol de liberação controlada (100 mg em 12h) durante 10 dias. Realizaram-se coletas de sangue venoso (5 mL), no período matutino, antes do tratamento e no 11º dia e as amostras foram armazenadas até análise (-70ºC). Foram utilizados testes enzimáticos ELISA para dosagem de citocinas plasmáticas (TNF-±, IL-1, IL-6) e receptores sTNF-R1, (R & D Systems). Realizou-se dosagem de citocinas em soro de 10 voluntários sadios. RESULTADOS: A concentração de TNF-± antes (5,8 ± 2,8 pg.mL-1) foi significativamente maior que após o tramadol (4,8 ± 2,1 pg.mL-1; p = 0,04, Teste Mann-Whitney). Não houve diferença significativa de IL-1², IL-6 e sTNF-R1 antes e após o tratamento. As concentrações plasmáticas de TNF-± (sadios: 1,4 ± 0,5; pacientes com dor: 5,8 ± 2,8 pg.mL-1; p = 0.01) e IL-6 (sadios: 1,2 ± 0,8; pacientes com dor: 3,5 ± 2,6 pg.mL-1; p = 0,01) foram significativamente maiores nos pacientes com dor neuropática que nos voluntários, Teste de Mann-Whitney. CONCLUSÕES: Nos pacientes com hérnia discal e síndrome do túnel do carpo as concentrações plasmáticas de TNF-± e IL-6 foram maiores que em voluntários sadios, não havendo diferença das concentrações de sTNF-R e IL-1². Houve redução da concentração plasmática de TNF-± após tratamento com tramadol (100 mg em 12h), mas não de IL-6, sTNF-R e IL-1².

BACKGROUND AND METHODS: Proinflammatory cytokines play an important role in the pathophysiology of neuropathic pain syndromes. The objective of this study was to evaluate plasma levels of proinflammatory cytokines before and after treatment with tramadol in patients with herniated intervertebral disks and carpal tunnel syndrome, and to compare them with normal individuals. METHODS: Thirty-eight patients with neuropathic pain secondary to herniated intervertebral disks or carpal tunnel syndrome participated in this study. All patients were treated with controlled release tramadol (100 mg every 12 hours) for 10 days. Venous blood (5 mL) was collected in the morning, before treatment and on the 11th day, and stored (-70° C) until analysis. ELISA was used to determine the plasma levels of cytokines (TNF-±, IL-1, IL-6) and receptors sTNF-R1 (R & D Systems). Plasma levels of cytokines of 10 healthy volunteers were also determined. RESULTS: The concentration of TNF-± before (5.8 ± 2.8 pg.mL-¹) was significantly higher than after treatment with tramadol (4.8 ± 2.1 pg.mL-1; p = 0.04, Mann-Whitney test). The levels of IL-1², IL-6, and sTNF-R1 before and after treatment with tramadol showed no significant differences. Plasma levels of TNF-± (healthy individuals: 1.4 ± 0.5; pain patients: 5.8 ± 2.8 pg.mL-1; p = 0.01) and IL-6 (healthy individuals: 1.2 ± 0.8; pain patients: 3.5 ± 2.6 pg.mL-1; p = 0.01) were significantly higher in patients with neuropathic pain, Mann-Whitney Test. CONCLUSIONS: In patients with herniated intervertebral disks and carpal tunnel syndrome, plasma levels of TNF-± and IL-6 were higher than in healthy volunteers, while differences in the concentrations of sTNF-R and IL-1² were not observed. Plasma levels of TNF-±, but not of IL-6, sTNF-R, and IL-1², decreased after treatment with tramadol (100 mg every 12 hours).

JUSTIFICATIVA Y OBJETIVOS: Las interleucinas proinflamatorias tienen una función importante en la fisiopatología de los síndromes dolorosos neuropáticos. El objetivo de este estudio, fue evaluar los niveles plasmáticos de interleucinas proinflamatorias antes y después del tratamiento con tramadol en pacientes con hernia de disco y síndrome del túnel del carpo, y compararlos con individuos normales. MÉTODO: Se investigaron 38 pacientes con dolor neuropático por hernia de disco o síndrome del túnel del carpo. Todos los pacientes fueron tratados con tramadol de liberación controlada (100 mg en 12h) durante 10 días. Se realizaron muestras de sangre venosa (5 mL), por la mañana, antes del tratamiento y en el 11º día, y las mismas se almacenaron para ser analizadas (-70ºC). Se utilizaron test enzimáticos ELISA para la dosificación de las interleucinas plasmáticas (TNF-±, IL-1, IL-6) y receptores sTNF-R1, (R & D Systems). Se realizó la dosificación de interleucinas en suero de 10 voluntarios sanos. RESULTADOS: La concentración de TNF-± antes (5,8 ± 2,8 pg.mL-1) fue significativamente mayor que después del tramadol (4,8 ± 2,1 pg.mL-1; p = 0,04, Test de Mann-Whitney). No hubo diferencia significativa de IL-1², IL-6 y sTNF-R1 antes y después del tratamiento. Las concentraciones plasmáticas de TNF-± (sanos: 1,4 ± 0,5; pacientes con dolor: 5,8 ± 2,8 pg.mL-1; p = 0.01) y IL-6 (sanos: 1,2 ± 0,8; pacientes con dolor: 3.5 ± 2,6 pg.mL-1; p = 0,01) fueron significativamente mayores en los pacientes con dolor neuropático que en los voluntarios, test de Mann-Whitney. CONCLUSIONES: En los pacientes con hernia discal y síndrome del túnel del carpo, las concentraciones plasmáticas de TNF-± y IL-6, fueron más elevadas que en los voluntarios sanos, no habiendo ninguna diferencia en las concentraciones de sTNF-R y IL-1². Hubo una reducción de la concentración plasmática de TNF-±; después del tratamiento con tramadol (100 mg en 12h), pero no de IL-6 sTNF-R y IL-1².

Asthmatics infected with Schistosoma mansoni have a less severe course of asthma and an inhibition of the Th2 inflammatory response that seems to be mediated by interleukin (IL-10). The objective of this study was to evaluate the capacity of some S. mansoni antigens to stimulate IL-10 production in vitro by cells of asthmatic infected individuals. Peripheral bloods mononuclear cells were stimulated with the S. mansoni recombinant antigens Sm22.6, Sm14, P24, and PIII antigen. IL-10 was measured in the supernatants of cultures. As the recombinant antigens were cloned in Escherichia coli, we blocked contaminant endotoxin with polymyxin B added to the cultures. We demonstrated that all antigens used drove high production of IL-10 in S. mansoni infected individuals (n = 13, 408 ± 514 and 401 ± 383 pg/ml, 484 ± 245 pg/ml, 579 ± 468 pg/ml, respectively). In asthmatics infected with S. mansoni (n = 21) rP24 induced higher levels of IL-10 (565 ± 377 pg/ml) when compared to PIII, rSm14 and rSm22.6 (184 ± 209 pg/ml; 292 ± 243 pg/ml; 156 ± 247 pg/ml, respectively). Conclusion: the S. mansoni antigens evaluated in this study stimulated IL-10 production by cells from infected individuals and therefore they have the potential to be used as a modulator of the inflammatory response in asthma.

Human T cell leukemia virus type-I (HTLV-I) infection is associated with spontaneous T cell activation and uncontrolled lymphocyte proliferation. An exacerbated type-1 immune response with production of pro-inflammatory cytokines (interferon-gamma and tumor necrosis factor-alpha) is significantly higher in patients with myelopathy associated to HTLV-I than in HTLV-I asymptomatic carriers. In contrast with HTLV-I, a chronic Schistosoma mansoni infection is associated with a type-2 immune response with high levels of interleukin (IL-4, IL-5, and IL-10) and low levels of IFN-gamma. In this study, clinical and immunological consequences of the HTLV-I and S. mansoni infection were evaluated. The immune response in patients with schistosomiasis co-infected with HTLV-I showed low levels of IL-5 (p < 0.05) in peripheral blood mononuclear cells cultures stimulated with S. mansoni antigen (SWAP) and decreased SWAP-specific IgE levels when compared with patients with only schistosomiasis (p < 0.05). Liver fibrosis was mild in all HTLV-I co-infected patients. Immunological response was also compared in individuals who had only HTLV-I infection with those who were co-infected with HTLV-I and helminths (S. mansoni and Strongyloides stercoralis). In patients HTLV-I positive co-infected with helminths the IFN-gamma levels were lower than in individuals who had only HTLV-I. Moreover, there were fewer cells expressing IFN-gamma and more cells expressing IL-10 in individuals co-infected with HTLV-I and helminths. These dates indicate that HTLV-I infection decrease type 2-response and IgE synthesis and are inversely associated with the development of liver fibrosis. Moreover, helminths may protect HTLV-I infected patients to produce large quantities of pro-inflammatory cytokines such as IFN-gamma.

This study objective was to evaluate the cytokines associated with early events of hepatic fibrosis in schistosomiasis mansoni. Hepatic fibrosis was classified by ultrasonography in 94 patients. Immunological evaluation was performed by measurement of secreted cytokines (interleukin IL-5, IL-10, IL-13, interferon-gamma, tumor necrosis factor-alpha and transforming growth factors-beta) in peripherl blood mononuclear cells stimulated by Schistosoma mansoni antigens. Significantly, higher levels of IL-5, IL-10 and IL-13 were found in supernatants of SEA-stimulated PBMC from subjects with degree III hepatic fibrosis as compared to patients with degree I or II fibrosis, Significant increases in IL-5 and IL-13 levels were also observed in some of the subjects who remained untreated for one year following initial assessment and developed more serious fibrosis during this period. The data suggests a role for type 2 cytokines in early stages of hepatic fibrosis in human schistosomiasis mansoni.

Chronic Schistosoma mansoni infection leads to a type 2-immune response with increased production of interleukin (IL-10). Evidence indicates chronic exposure to S. mansoni down regulates the type 1 immune response and prevents the onset of Th1-mediated diseases such as multiple sclerosis, diabetes mellitus and Cronh's disease. Furthermore, our own studies have revealed that chronic exposure to S. mansoni also down regulates atopic disease, Th2-mediated diseases. Our studies show an inverse association between the skin prick test reactivity and infection with S. mansoni and show the severity of asthma is reduced in subjects living in an endemic area of S. mansoni. Moreover, we hypothesize the mechanisms involved in the modulation of inflammatory response in atopic individuals, is likely dependent on IL-10 production, an anti-inflammatory cytokine elevated during helminth infections. Patients with asthma and helminth infections produced less IL-5 than patients with asthma without helminth infections, and this down regulation could, in part, be mediated by IL-10. In conclusion, helminthic infections, through induction of regulatory mechanisms, such as IL-10 production, are able to modulate the inflammatory immune response involved in the pathology of auto-immune and allergic disease.

Realizou-se a análise comparativa de custos de produção de feijão, após um ano em sistemas de plantio direto e convencional, no período de inverno, com irrigação por pivô central. Os experimentos foram desenvolvidos na Fazenda Dois Irmãos, em Pereira Barreto (SP), em delineamento experimental de blocos ao acaso, com 11 tratamentos e quatro repetições, para cada sistema de plantio. Os tratamentos constaram de uma testemunha sem N, duas fontes de N (uréia e nitrato de amônio) e cinco dosagens (20, 40, 60, 80 e 100 kg.ha-1 de N), aplicados em cobertura aos 27 dias após a emergência das plantas. Utilizou-se densidade de 15 plantas por metro de linha e 0,50 m entre sulcos, com semeadura realizada em 20/5/2001. Na área do sistema de plantio direto, as plantas daninhas foram dessecadas após o cultivo do milho, e na de plantio convencional, realizaram uma aração e duas gradagens, sendo uma pesada e uma niveladora, no preparo do solo. Durante o desenvolvimento da cultura foram aplicados herbicidas em pós-emergência para o controle do mato. As duas fontes de nitrogênio mostraram efeito positivo na produção de grãos. A análise econômica revelou melhor resultado para uréia, na dose de 60 kg.ha-1 de N, em preparo convencional do solo, com índice de lucratividade de 40,6%. Para o plantio direto, o nitrato de amônio, na base de 40 kg.ha-1 de N, mostrou maior acréscimo na receita e lucratividade de 25,6%.

A comparative economical evaluation of conventional and no-tillage bean crop systems was done. A field experiment was settled under irrigation, at Dois Irmãos Farm, Pereira Barreto, São Paulo State, sowed in the winter of 2001. The experimental design was a randomized block with four replications for each tillage system. N fertilization was applied 27 days after emergency at the rates of 20, 40, 60, 80, 100 kg.ha-1 of N. Urea and ammonium nitrate were used as nitrogen source. The cv. Carioca was used at a seed density of 15 seeds per meter in rows 0,50 cm apart. Herbicide was applied in the no-tillage plots before sowing. Conventional tillage plots were prepared with plowing and harrowing twice. The results show that bean crop responds differently the nitrogen source. The economical evaluation pointed out the higher profitable index of 40,6% for urea source at N doses of 60 kg.ha-1, under conventional tillage system. The best result for no tillage sowed 25,6% profitable index and was obtained with 40 kg.ha-1 of N.

In areas of Leishmania chagasi transmission the ability to control leishmania infection is associated with IFN-g production. In visceral leishmaniasis down-regulation of T cell responses is mediated by interleukin-10 (IL-10). In this study we evaluated the lymphoproliferative response, IFN-g and IL-10 production on lymphocyte cultures stimulated with recombinant leishmania antigens in subjects with asymptomatic L. chagasi infection. There was a statistically significant difference in the lymphoproliferative response of the subjects with asymptomatic infection as compared to patients with visceral leishmaniasis and healthy subjects with respect to crude antigens (p<0.01), gp-63 (p<0.05) and hsp-70 (p<0.01), as well as between asymptomatic L. chagasi infected subjects and patients with visceral leishmaniasis with respect to the response to all antigens tested. The IFN-g production observed in the group with asymptomatic infection with all the three recombinant antigens tested was higher (p<0.01) than that observed in patients with visceral leishmaniasis and in healthy subjects. Furthermore, in individuals with asymptomatic infection, IL-10 levels in cultures stimulated with recombinant antigens were very low. This study shows that lymphocytes from individuals with asymptomatic L. chagasi infection are able to recognize recombinant leishmania antigens with production of a cytokine that is associated with leishmania killing.

The frequency of asymptomatic infection among relatives and neighbors of cases of visceral leishmaniasis (VL) was compared and characterization of the immunological response in these subjects was performed. Cases were from a new endemic area, close to the beach and near Salvador, capital of the State of Bahia, Brazil. The characterization of asymptomatic infection was made using a skin reaction test and detection of antibody to Leishmania chagasi by the ELISA test. To characterize the immunological response of these subjects with asymptomatic L. chagasi infection the cytokines profile and the lymphoproliferative response were determined after stimulation of lymphocytes by L. chagasi antigen. There was no difference in the frequency of L. chagasi infection in relatives (45%) and in neighbors (27%) of cases of VL (P>0.05). The immunological response from these subjects was characterized by high production of IFN-g and a low production of IL-10 and a good lymphoproliferative response to L. chagasi antigen