Morphological analysis and lipid composition of the stable fly Stomoxys calcitrans salivary glands reinforce important features of feeding behaviour.

Stomoxys calcitrans L. (Diptera: Muscidae), the stable fly, is a hematophagous insect of great veterinary importance, because it is a mechanical vector of diverse pathogens in livestock. The saliva of blood-feeding insects presents important pharmacologically active molecules that impair blood clotting, promote vasodilation and modulate the host immune system response, crucial processes for successful feeding. These properties also enable pathogens' transmission. In the present work, we describe an efficient protocol to dissect S. calcitrans salivary glands, their morphological characteristics and lipid profile. The mean length of the tubular gland is 3.23 mm with a bulbous posterior end and a narrow anterior end. Histological analysis revealed a monolayer of large polygonal epithelial cells with voluminous nuclei and high lipid content in their cytoplasm. Ultrastructural analysis showed that the epithelium is rich in mitochondria, free ribosomes, Golgi complex cisternae, presenting a great extension of rough endoplasmic reticulum that contains an electron-dense material. Lipid analysis by thin-layer chromatography showed that neutral fatty acids and phosphatidylcholine are predominant in the fly salivary glands. Lysophosphatidylcholine, an important signalling biomolecule involved in different metabolic processes, including host's immunomodulation and pathogens proliferation and differentiation, is also present.

Stomoxys calcitrans L. (Diptera: Muscidae), a mosca­dos­estábulos, é um inseto hematófago de grande importância veterinária, uma vez que é vetor mecânico de diversos patógenos que infectam animais da pecuária. A saliva de insetos que se alimentam de sangue apresenta importantes moléculas farmacologicamente ativas que impedem coagulação sanguínea, promovem vasodilatação e modulam o sistema imune do hospedeiro, processos cruciais para uma alimentação bem sucedida. Tais propriedades também permitem a transmissão de patógenos. No presente trabalho, nós descrevemos um protocolo eficiente para dissecar as glândulas salivares de S. calcitrans, suas características morfológicas e perfil lipídico. O comprimento médio da glândula tubular é 3.23 mm com uma porção posterior bulbosa e porção anterior estreita. Análises histológicas revelaram uma monocamada de células epiteliais largas e poligonais com núcleos volumosos e alto conteúdo lipídico em seus citoplasmas. Análises ultraestruturais mostraram um epitélio rico em mitocôndria, ribossomos livres, cisternas do complexo de Golgi, apresentando uma grande extensão de retículo endoplasmático que contém um material eletrodenso. A análise lipídica mostrou que ácidos graxos neutros e fosfatidilcolina predominam nas glândulas salivares da mosca. Lisofosfatidilcolina, uma importante biomolécula sinalizadora envolvida em diferentes processos metabólicos, incluindo imunomodulação do hospedeiro e proliferação e diferenciação de patógenos, também se encontra presente.

Dietary olive oil intake aggravates psoriatic skin inflammation in mice via Nrf2 activation and polyunsaturated fatty acid imbalance.

Psoriasis is a chronic inflammatory skin disease, which does not have effective treatment options. However, olive oil has been suggested as an alternative to treat psoriasis, but no study has evaluated the mechanisms involved in the effects of olive oil on psoriasis. Thus, the current study investigated whether olive oil could ameliorate psoriasiform skin inflammation. To test this, mice received topical application of imiquimod to induce inflammation and were treated orally with olive oil. Human immortalized keratinocytes were also treated with imiquimod and olive oil. Epidermal thickness and keratinocyte proliferation were increased in imiquimod-induced lesions of olive-oil-treated animals. In both in vitro and in vivo studies, protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) were elevated following imiquimod and olive oil administration. Inhibition of Nrf2 abolished the increased proliferation of keratinocytes treated with imiquimod and olive oil, demonstrating the role of Nrf2 in olive oil-mediated exacerbation of psoriasiform skin inflammation. In addition, lower levels of linoleic acid and higher levels of oleic acid were observed in imiquimod- and olive-oil-treated animals, which may also contribute to the adverse effects of olive oil on psoriasis. In conclusion, dietary intake of olive oil aggravates the symptoms of psoriatic skin lesions through the overexpression of Nrf2 and an imbalance in oleic and linoleic acids levels, suggesting that a diet rich in olive oil may have significant negative effects on psoriasis.

Development of a ligand blot assay using biotinylated live cells.

Adhesive interactions between cells are critical to a variety of processes, including host-pathogen relationships. The authors have developed a new technique for the observation of binding interactions in which molecules obtained from excised tissues are resolved by gel electrophoresis and transferred to a membrane. Biotinylated live cells are then kept in contact with that membrane, and their interactions with proteins of interest are detected by peroxidase-labeled streptavidin, followed by a biotin-streptavidin detection system. The adhesion proteins can eventually be identified by cutting the relevant band(s) and performing mass spectrometry or other amino acid-sequencing methods. The technique described here allows for the identification of both known and novel adhesion molecules capable of binding to live cells, among a complex mixture and without previous isolation or purification. This is especially important for the analysis of host-parasite interactions and may be extended to other types of cell-cell interactions.