Abstract Although there are reports that polyphenol resveratrol (Rsv) may cause muscle hypertrophy in basal conditions and attenuate muscle wasting in catabolic situations, its mechanism of action is still unclear. Our study evaluated the ex vivo effects of Rsv on protein metabolism and intracellular signaling in innervated (sham-operated; Sham) and 3-day sciatic denervated (Den) rat skeletal muscles. Rsv (10-4 M) reduced total proteolysis (40%) in sham muscles. Den increased total proteolysis (~40%) in muscle, which was accompanied by an increase in the activities of ubiquitin-proteasome (~3-fold) and lysosomal (100%) proteolytic systems. Rsv reduced total proteolysis (59%) in Den muscles by inhibiting the hyperactivation of ubiquitin-proteasome (50%) and lysosomal (~70%) systems. Neither Rsv nor Den altered calcium-dependent proteolysis in muscles. Mechanistically, Rsv stimulated PKA/CREB signaling in Den muscles, and PKA blockage by H89 (50μM) abolished the antiproteolytic action of the polyphenol. Rsv reduced FoxO4 phosphorylation (~60%) in both Sham and Den muscles and Akt phosphorylation (36%) in Den muscles. Rsv also caused a homeostatic effect in Den muscles by returning their protein synthesis rates to levels similar to Sham muscles. These data indicate that Rsv directly inhibits the proteolytic activity of lysosomal and ubiquitin-proteasome systems, mainly in Den muscles through, at least in part, the activation of PKA/CREB signaling. (Rsv situations unclear shamoperated operated (sham-operated 3day day 3 (Den 104 10 4 (10- M 40% 40 (40% ~40% (~40% ubiquitinproteasome ubiquitin proteasome ~3fold 3fold fold ~3 (~3-fold 100% 100 (100% systems 59% 59 (59% 50% 50 (50% ~70% 70 (~70% calciumdependent calcium dependent Mechanistically PKACREB CREB H H8 50μM μM (50μM FoxO ~60% 60 (~60% 36% 36 (36% through part 1 (10 (40 ~40 (~40 ~ (100 5 (59 (50 ~70 7 (~70 ~60 6 (~60 (36 (1 (4 ~4 (~4 (5 ~7 (~7 ~6 (~6 (3 ( (~

Abstract Cultivation of species of the genus Eucalyptus is important for the Brazilian economy, with 6.97 million hectares planted. Glycaspis brimblecombei Moore (Hemiptera: Aphalaridae), detected in Brazil in 2003, has dispersed and now damages Eucalyptus crops in all regions of this country. The location and identification of entomopathogenic fungi isolates may increase the options for integrated pest management. The objective of this research was to evaluate the pathogenicity of Cordyceps cateniannulata and Cordyceps javanica isolates to G. brimblecombei. Ten nymphs of G. brimblecombei, with or without lerps, were placed per Eucalyptus leaf cut with one of its edges on hydroretentive gel inside Petri dishes. The fungi isolates were suspended in a solution of Tween 80 (0.1%) at the concentration of 1.0 x 108 conidia mL-1 and sprayed on the G. brimblecombei nymphs. The mortality of this insect was evaluated daily for seven days, and the dead individuals were transferred to humid chambers. The conidia viability of the isolates was greater than 93%. The mortality of G. brimblecombei nymphs, seven days after the application of the fungi, was 100%. This is the first report of the pathogenicity of C. cateniannulata and C. javanica isolates, occurring naturally in the field, to G. brimblecombei.

Abstract Syzygium malaccense is popularly used to treat inflammation and pain-related ailments. The species was assessed regarding its antioxidant, antiglycant, anti-inflammatory, including anti-neuroinflammatory, and antinociceptive activities. Different models were employed to measure S. malaccense extract (ESM) antioxidant activity. The antiglycant activity was determined using the glucose-induced protein glycation model. LPS-induced neuroinflammation on murine BV-2 microglial cell line was used for anti-neuroinflammatory activity evaluation. The croton oil-induced ear edema test was accomplished to evaluate the in vivo anti-inflammatory activity. Acetic acid-induced writhing together with formalin-induced paw licking assays were performed to evaluate the antinociceptive potential. Finally, the chemical characterization was accomplished by a UHPLC-MS analysis. ESM presented relevant antioxidant and antiglycant activity. NO production by BV-2 cells was reduced, indicating the relevant neuroprotective activity. ESM significantly decreased the mice ear edema induced by croton oil and the nociceptive stimulus induced by acetic acid and formalin by central and peripheral mechanisms. The flavonoids myricitrin, myricetin and quercetin were identified and, as far as we know, the alkaloid reserpine was reported in the species for the first time. The antioxidant and antiglycant potential of ESM, may be related to the in vivo anti-inflammatory and antinociceptive effects, and to the in vitro neuroinflammation inhibition.

RESUMO: O ácido 2-cloroetilfosfônico (ethephon) e o carbureto de cálcio são usados para induzir e sincronizar a floração da cultura do abacaxi. Alguns produtores têm utilizado, em associação com esses indutores do florescimento, o herbicida 2,4-D. Essa prática baseia-se na hipótese de que o 2,4-D atua como regulador de crescimento, aumentando o tamanho e peso dos frutos. Diante disso, os objetivos deste trabalho foram verificar se a utilização do 2,4-D, associado ao carbureto de cálcio ou ao ethephon, influencia a produtividade e qualidade dos frutos do abacaxizeiro e, também, se o uso desse herbicida deixa resíduos nos frutos. Para isso, foram realizados dois experimentos de campo em duas fazendas. Os tratamentos consistiram da aplicação de ethephon e carbureto de cálcio, isoladamente e em mistura com duas, quatro e seis gotas de 2,4-D no centro da roseta foliar do abacaxizeiro cv. Pérola aos 10 meses após o seu plantio. O uso de 2,4-D em mistura com o carbureto de cálcio ou o ethephon, independentemente da dose e do ambiente de cultivo, não influenciou o tamanho, peso, teor de sólidos solúveis totais, densidade e pH dos frutos de abacaxi. Todavia, dependendo do ambiente de cultivo, as misturas de ethephon + 2,4-D não influenciaram ou reduziram o florescimento e a produtividade da cultura. Não foram encontrados resíduos do 2,4-D na polpa dos frutos do abacaxi. Concluiu-se que o uso de 2,4-D associado ao carbureto de cálcio ou ao ethephon como regulador de crescimento do abacaxi, nas condições avaliadas, não incorpora benefícios qualitativos ou quantitativos ao sistema produtivo dessa cultura. Além disso, não foram encontrados resíduos de 2,4-D na polpa dos frutos de abacaxi.

ABSTRACT: The 2-Chloroethylphosphonic acid (ethephon) and calcium carbide are used to induce and synchronize the flowering of pineapple crop. Some farmers have used the 2,4-D herbicide in combination with these flowering inducers. This practice is based on the hypothesis that 2,4-D would act as a growth regulator, increasing fruit size and weight. The objectives of this work was to verify if the use of 2,4-D, associated to calcium carbide or ethephon, influences the productivity and quality of the pineapple fruits and, also if the use of this herbicide leaves residues in the fruits. For this, two field experiments were carried out in two different farms. The treatments consisted of the application of ethephon and calcium carbide, alone, and in a mixture with two, four and six drops of 2,4-D in the center of the leaflet of the pineapple cv. “Pérola” at 10 months after planting. The use of 2,4-D mixed with calcium carbide or ethephon, regardless of the dose and culture environment, did not influence the size, weight, total soluble solids content, density and pH of the pineapple fruits. However, depending on the growing environment, ethephon + 2,4-D mixtures did not influence or reduce flowering and crop yield. No 2,4-D residues were found in the fruit pulp of pineapple. It was concluded that the use of 2,4-D as a growth regulator of pineapple under the conditions evaluated does not incorporate qualitative or quantitative benefits to the productive system of this crop. In addition, no 2,4-D residues were found in the fruit pulp of pineapple.

RESUMO: A resposta imune da glândula mamária desempenha um papel importante ao determinar o estabelecimento da infecção. Desta forma, a melhor compreensão da função dos neutrófilos irá nos subsidiar conhecimentos, pelo qual podemos evitar o processo inflamatório pela otimização da resposta imune de bovinos leiteiros, e fornece ferramentas para diagnosticar a mastite ou um possível instrumento para identificar e selecionar animais resistentes à infecção intramamária, aumentando a produtividade do rebanho. O presente estudo utilizou 112 amostras provenientes de quartos mamários de 28 vacas Holandesas que foram divididos em amostras de leite com baixa (n=72; <2×105 células mL-1) ou alta (n=40; 2×105 células mL-1) contagem de células somáticas (CCS). A porcentagem de neutrófilos no leite, a produção intracelular de espécies reativas de oxigênio (ERO) e a fagocitose de Staphylococcus aureus pelos neutrófilos do leite foram avaliadas por citometria de fluxo. Os resultados do presente estudo demonstraram maior percentagem de neutrófilos (CH138+; P=0,0003) e percentagem de neutrófilos que produziram ERO (P=0,008) em amostras de leite com alta CCS. Por outro lado, a intensidade de fagocitose de S. aureus por neutrófilos em amostras de leite com baixa CCS (P=0,003), que demonstra maior atividade funcional destas células neste grupo. As funções neutrofílicas para o diagnóstico da mastite não apresentaram valores de sensibilidade e especificidade altos, que foram confirmados pelo índice Youden. Desta forma, conclui-se que a produção intracelular de ERO e fagocitose de S. aureus pelos neutrófilos do leite não apresentaram valores preditivos altos para detecção de mastite. Além disto, os resultados do presente estudo reforçam a ideia de neutrófilos do leite com menor capacidade fagocítica podem ser associados à alta CCS, e pode ser considerado como uma ferramenta para identificar animais mais susceptíveis à infecções intramamárias.

ABSTRACT: The immune response capacity of the mammary gland plays a major role to determine if mastitis will or not be established. Thus, we hypothesize that a better understanding of polymorphonuclear neutrophil leukocyte (PMN) function will elucidate mechanisms that will improve our knowledge of how we could avoid an inflammatory process by increasing the immune capacity of the cow, and even further, to search for a tool to diagnose mastitis or a possible way to select and identify non-susceptible animals. The present study utilized 112 quarters from 28 Holstein dairy cows that were divided into quarters milk samples with somatic cell count (SCC) <2×105 cells mL-1 (n=72) and SCC >2×105 cells mL-1 (n=40). The percentages of milk PMNs and the levels of intracellular reactive oxygen species (ROS) and the phagocytosis of Staphylococcus aureus by milk neutrophils were evaluated by flow cytometry. Our results showed a higher percentage of neutrophils in quarter milk samples with high SCC (P=0.0003), and this group also had a significantly higher percentage of neutrophils that produced ROS (P=0.008). On the other hand, the phagocytosis intensity of S. aureus by milk neutrophils was higher in quarters with low SCC (P=0.003), suggesting a better mammary gland immunity against invading pathogens. Analyzing the results of the predictive values of the measured PMN functions, they cannot be used isolated as a good diagnosis test since none of them had a satisfactory sensitivity and specificity values, which was also confirmed by the Youden index values being far from one. In conclusion, the assessment of milk bovine neutrophil functions could improve our understanding of the cellular basis of mastitis. Although, the intracellular ROS production and S. aureus phagocytosis by milk neutrophil did not have high predictive values to detect intramammary infections, our results strengthen the idea that that poor bovine mammary gland neutrophil phagocytic ability may be associated with high SCC, and might be considered to identify susceptible dairy cows to mastitis.

OBJETIVO: Analisar a ocorrência de anemia e de deficiência de ferro em crianças de 1 a 5 anos e a associação destes desfechos com a deficiência de retinol. MÉTODOS: Trata-se de um estudo observacional analítico do tipo transversal, realizado no município de Vitória - ES, entre abril e agosto de 2008, com crianças (n = 692) saudáveis de 1 a 5 anos, residentes em áreas de abrangência de Unidades Básicas de Saúde. Foram avaliados dados sociodemográficos, econômicos, dietéticos (ingestão de energia, proteína, ferro e vitamina A), antropométricos (índice de massa corporal-por-idade e estatura-por-idade) e bioquímicos (níveis séricos de ferritina, hemoglobina e retinol). RESULTADOS: Detectou-se anemia, deficiência de ferro e deficiência de retinol em 15,7%, 28,1% e 24,7% das crianças, respectivamente. A análise univariada evidenciou maior ocorrência de anemia (RP: 4,62; IC 95%: 3,36; 6,34, p < 0.001) e de deficiência de ferro (RP: 4,51; IC 95%: 3,30; 6,17, p < 0.001) entre crianças que apresentavam deficiência de retinol. As mesmas relações se mantiveram após o ajuste pelas variáveis socioeconômicas, demográficas, dietéticas e antropométricas. Houve relação positiva entre os valores de ferritina sérica vs. retinol (r = 0,597; p < 0,001) e hemoglobina vs. retinol (r = 0,770; p < 0,001). CONCLUSÕES: A anemia e a deficiência de ferro mostraram-se associadas com baixos níveis de retinol em crianças de 1 a 5 anos, e houve correlação positiva dos níveis de retinol com os de ferritina sérica e hemoglobina. Isto torna importante iniciativas que estimulem o desenvolvimento de novos tratamentos e a ampliação de pesquisas em relação à deficiência de retinol.

OBJECTIVE: To analyze the occurrence of anemia and iron deficiency in children aged 1 to 5 years and the association of these events and retinol deficiency. METHODS: This was an observational analytic cross-sectional study conducted in Vitoria, ES, Brazil, between April and August of 2008, with healthy children aged 1 to 5 years (n = 692) that lived in areas covered by primary healthcare services. Sociodemographic and economic conditions, dietary intake (energy, protein, iron, and vitamin A ingestion), anthropometric data (body mass index-for-age and height-for-age), and biochemical parameters (ferritin, hemoglobin, and retinol serum) were collected. RESULTS: The prevalence of anemia, iron deficiency, and retinol deficiency was 15.7%, 28.1%, and 24.7%, respectively. Univariate analysis showed a higher prevalence of anemia (PR: 4.62, 95% CI: 3.36, 6.34, p < 0.001) and iron deficiency (PR: 4.51, 95% CI: 3.30, 6.17, p < 0.001) among children with retinol deficiency. The same results were obtained after adjusting for socioeconomic and demographic conditions, dietary intake, and anthropometric variables. There was a positive association between ferritin vs. retinol serum (r = 0.597; p < 0.001) and hemoglobin vs. retinol serum (r = 0.770; p < 0.001). CONCLUSIONS: Anemia and iron deficiency were associated with low levels of serum retinol in children aged 1 to 5 years, and a positive correlation was verified between serum retinol and serum ferritin and hemoglobin levels. These results indicate the importance of initiatives encouraging the development of new treatments and further research regarding retinol deficiency.

In Brazil, the presence of plasmid-mediated AmpC (pAmpC)-producing isolates has been sporadically reported. We evaluated the frequency of pAmpC among 133 Enterobacteriaceae clinical isolates. The blaCMY-2-like gene was detected in a single Klebsiella pneumoniae isolate. In our study, the pAmpC frequency was very low as previously reported.

Understanding the membrane solubilization process and finding effective solubilizing agents are crucial challenges in biochemical research. Here we report results on the interaction of the novel linear alkylamido propyl dimethyl amino propanosulfonate detergents, ASB-14 and ASB-16, with human erythrocyte membranes. An estimation of the critical micelle concentration of these zwitterionic detergents (ASB-14 = 100 µM and ASB-16 = 10 µM) was obtained using electron paramagnetic resonance. The amount of proteins and cholesterol solubilized from erythrocytes by these detergents was then determined. The hemolytic activities of the ASB detergents were assayed and the detergent/lipid molar ratios for the onset of hemolysis (Re sat) and total lysis (Re sol) were calculated, allowing the determination of the membrane binding constants (Kb). ASB-14 presented lower membrane affinity (Kb = 7050 M-1) than ASB-16 (Kb = 15610 M-1). The amount of proteins and cholesterol solubilized by both ASB detergents was higher while Re sat values (0.22 and 0.08 detergent/lipid for ASB-14 and ASB-16, respectively) were smaller than those observed with the classic detergents CHAPS and Triton X-100. These results reveal that, besides their well-known use as membrane protein solubilizers to enhance the resolution of two dimensional electrophoresis/mass spectrometry, ASB-14 and ASB-16 are strong hemolytic agents. We propose that the physicochemical properties of ASB detergents determine their membrane disruption efficiency and can help to explain the improvement in the solubilization of membrane proteins, as reported in the literature.

A família Bromeliaceae, dentre a grande variedade de plantas tropicais nativas do Brasil, tem merecido destaque devido à sua importância econômica como plantas ornamentais, sendo atualmente muito cultivadas e utilizadas na decoração de interiores e em projetos paisagísticos. Alguns gêneros são endêmicos da Floresta Atlântica e, em função dessa procura, a retirada de seus ambientes naturais constitui ameaça a algumas espécies. A Unidade de Pesquisa e Conservação de Bromeliaceae (UPCB), localizada na Universidade Federal de Viçosa (UFV), MG, tem como finalidade promover a pesquisa em favor da conservação da família Bromeliaceae. Um problema constante na manutenção desse acervo é a infestação por plantas daninhas. Objetivou-se neste trabalho definir as espécies de plantas daninhas críticas no cultivo de bromélias. Foram realizadas visitas semanais, no período de 17 de novembro de 2006 a 17 de janeiro de 2007, para caracterização do comportamento das espécies de plantas daninhas no cultivo de bromélias na UPCB. Após esse período, realizou-se capina manual dos vasos e triagem das espécies daninhas, que foram identificadas e quantificadas. As espécies críticas foram descritas e seus indivíduos férteis depositados no Herbário VIC, do Departamento de Biologia Vegetal, como material testemunha. Realizou-se, também, documentação fotográfica das espécies durante o período de infestação. Foram identificadas duas espécies críticas: brilhantina (Pilea microphylla), com média de seis indivíduos por vaso, e agriãozinho (Cardamine bonariensis), com média de 13 indivíduos por vaso; sete espécies consideradas potencialmente críticas, com destaque para barba-de-falcão (Crepis japonica); e 12 espécies oportunistas.

The Bromeliaceae family stands out among the great variety of Brazilian native tropical plants because of the economic importance of its ornamental plants, currently widely cultivated and used for indoors decorations and landscape projects. Some genera are endemic to the Atlantic Forest and to meet their great demand, removal from their natural environment has become a threat to some species. The Research and Conservation of Bromeliaceae (UPCB) Unit of the Universidade Federal de Vicosa, in Vicosa, MG promotes research on Bromeliaceae conservation. Weed infestation is a constant problem in the maintenance of this collection. The objective of this work was to define the most critical weeds in bromeliad cultivation. From November 2006 to January 2007, weekly visits were conducted to the collection of bromeliads planted in pots at the UPCB, aiming to characterize the occurrence and behavior of weed species. After this period, the weeds were manually removed from the pots, identified and quantified. The critical weed species were described and their fertile individuals were included in the VIC Herbarium of the Department of Plant Biology, as checks. Photographic documentation of the weeds was also conducted. Two critical species were identified: Pilea microphylla,with around six individuals per pot, and Cardamine bonariensis, with approximately 13 individuals per pot. Seven other species were considered potentially critical, with Crepis japonica being the most important one. Twelve opportunistic species were also identified.

Philander frenata e Didelphis marsupialis mantém Trypanosoma cruzi sem desenvolver aparentemente nenhuma doença. Ao contrário de Didelphis marsupialis, Philander frenata mantém subpopulações do grupo T. cruzi II. Aqui, nós comparamos a resposta imune humoral de dois didelfídeos natural e experimentalmente infectados com o grupo T. cruzi II, empregando as técnicas de SDS-PAGE/Western blot e por um método de imunofluorescência indireta. Também estudamos os padrões histopatológicos de P. frenata infectado natural e experimentalmente por T. cruzi. O soro de P. frenata reconheceu mais antígenos que o de D. marsupialis. Polipeptídeos de 66 e 90kDa foram os antígenos mais reconhecidos por ambas espécies nas frações enriquecidas de membranas solúveis. P. frenata reconheceu intensamente o antígeno de 45kDa. Nossos resultados indicam que: não existem diferenças quantitativas e qualitativas nas fases subpatente e patente no padrão de reconhecimento de P. frenata; as diferenças no padrão de reconhecimento de antígenos de parasitas pelos soros de P. frenata e D. marsupialis sugerem que provavelmente essas espécies "aprenderam" a viver em harmonia com T. cruzi por estratégias distintas; embora P. frenata não apresente doença aparente, lesões tissulares tendem a ser mais severas do que as descritas para D. marsupialis; ambos didelfídeos provavelmente adquiriram a infecção por T. cruzi depois de sua divergência evolucionária.

Philander frenata and Didelphis marsupialis harbor parasitism by Trypanosoma cruzi without developing any apparent disease and on the contrary to D. marsupialis, P. frenata maintains parasitism by T. cruzi II subpopulations. Here we compared the humoral immune response of the two didelphids naturally and experimentally infected with T. cruzi II group, employing SDS-PAGE/Western blot techniques and by an Indirect immunofluorescence assay. We also studied the histopathological pattern of naturally and experimentally infected P. frenata with T. cruzi. P. frenata sera recognized more antigens than D. marsupialis, and the recognition pattern did not show any change over the course of the follow up of both didelphid species. Polypeptides of 66 and 90kDa were the most prominent antigens recognized by both species in the soluble and enriched membrane fractions. P. frenata recognized intensely also a 45kDa antigen. Our findings indicate that: 1) there were no quantitative or qualitative differences in the patent or subpatent phases in the recognition pattern of P. frenata; 2) the significant differences in the recognition pattern of parasitic antigens by P. frenata and D. marsupialis sera suggest that they probably "learned" to live in harmony with T. cruzi by different strategies; 3) although P. frenata do not display apparent disease, tissular lesions tended to be more severe than has been described in D. marsupialis; and 4) Both didelphids probably acquired infection by T. cruzi after their evolutionary divergence.

Nascent procollagen peptides and other secretory proteins are transported across the endoplasmic reticulum (ER) membrane through a protein-conducting channel called translocon. Sec61alpha, a multispanning membrane translocon protein, has been implicated as being essential for translocation of polypeptide chains into the cisterns of the ER. Sec61alpha forms a protein complex with collagen and Hsp47, an ER-resident heat shock protein that binds specifically to collagen. However, it is not known whether Sec61alpha is ubiquitously produced in collagen-producing F9 teratocarcinoma cells or under heat shock treatment. Furthermore, the production and utilization of Sec61alpha may depend on the stage of cell differentiation. Cultured F9 teratocarcinoma cells are capable of differentiation in response to low concentrations of retinoic acid. This differentiation results in loss of tumorigenicity. Mouse F9 cells were grown in culture medium at 37ºC and 43ºC (heat shock treatment) treated or not with retinoic acid, and labeled in certain instances with 35S-methionine. Membrane-bound polysomes of procollagen IV were then isolated. Immunoprecipitation and Western blot analysis were performed using polyclonal antibodies against collagen IV, Hsp47 and Sec61alpha. Under retinoic acid-untreated conditions, F9 cells produced undetectable amounts of Sec61alpha. Sec61alpha, Hsp47 and type IV collagen levels were increased after retinoic acid treatment. Heat shock treatment did not alter Sec61alpha levels, suggesting that Sec61alpha production is probably not affected by heat shock. These data indicate that the enhanced production of Sec61alpha in retinoic acid-induced F9 teratocarcinoma cells parallels the increased synthesis of Hsp47 and collagen type IV.