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.

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.

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.