The integrin family not only mediates the recruitment of polymorphonuclear leukocytes (PMN) to sites of inflammation but also regulates several effector functions by binding to specific ligands. We have recently demonstrated that soluble fibrinogen (sFbg) is able to trigger an activating signal in PMN through an integrin-dependent mechanism. This activation results in degranulation, phagocytosis enhancement, and apoptosis delay. The aim of the present work was to further elucidate the molecular events that follow sFbg interaction with CD11b in human PMN, and the participation of this signaling pathway in the regulation of neutrophil functionality. We demonstrate that sFbg triggers a cascade of intracellular signals that lead to focal adhesion kinase and extracellular signal-regulated kinase 1/2 tyrosine phosphorylation. The activation of this mitogen-activated protein kinase pathway plays a central role in the sFbg modulation of secondary granule degranulation, Ab-dependent phagocytosis, and apoptosis. However, fibrinogen-induced secretory vesicle degranulation occurs independently of the signaling transduction pathways investigated herein. In the context of an inflammatory process, the intracellular signal pathway activated by sFbg may be an early event influencing the functionality of PMN.

SummaryHaemolytic uraemic syndrome (HUS) is caused by Shiga-toxin-producing Escherichia coli (STEC). Although, Shiga toxin type 2 (Stx2) is responsible for the renal pathogenesis observed in patients, the inflammatory response, including cytokines and polymorphonuclear neutrophils (PMN), plays a key role in the development of HUS. Previously, we demonstrated that Stx2 injection generates an anti-inflammatory reaction characterized by endogenous glucocorticoid (GC) secretion, which attenuates HUS severity in mice. Here, we analysed the effects of Stx2 on the pathogenic function of PMN and the potential role of endogenous GC to limit PMN activation during HUS development in a murine model. For this purpose we assessed the functional activity of isolated PMN after in vivo treatment with Stx2 alone or in simultaneous treatment with Ru486 (GC receptor antagonist). We found that Stx2 increased the generation of reactive oxygen intermediates (ROI) under phobol-myristate-acetate (PMA) stimulation and that the simultaneous treatment with Ru486 strengthened this effect. Conversely, both treatments significantly inhibited in vitro phagocytosis. Furthermore, Stx2 augmented in vitro PMN adhesion to fibrinogen (FGN) and bovine serum albumin (BSA) but not to collagen type I (CTI). Stx2 + + + + Ru486 caused enhanced adhesion to BSA and CTI compared to Stx2. Whereas Stx2 significantly increased migration towards N -formyl-methionyl-leucyl-phenylalanine (fMLP), Stx2 + + + + Ru486 treatment enhanced and accelerated this process. The percentage of apoptotic PMN from Stx2-treated mice was higher compared with controls, but equal to Stx2 + + + + Ru486 treated mice. We conclude that Stx2 activates PMN and that the absence of endogenous GC enhances this activation suggesting that endogenous GC can, at least partially, counteract PMN inflammatory functions.

Molecular characterization of Giardia duodenalis in African countries is relatively scarce. The global understanding of Giardia epidemiology is reinforced when more data are available from highly endemic countries. In the present study, 50 fecal samples collected from children from Guinea-Bissau were screened for Giardia infection. Amplification of the Giardia ssu-rRNA fragment was achieved for 28 samples (28/50, 56.0 %) of which 23 (23/50, 46.0 %) positive samples for Giardia were detected through microscopy. Eighteen samples previous amplified for the ssu-rRNA locus were amplified for the bg gene fragment. Sequence analysis was performed in 26 and 17 samples for the ssu-rRNA and bg gene fragment, respectively. Our results revealed a predominance of assemblage B (22/26, 84.6 %), sequences with high genetic polymorphism among isolates belonging to this assemblage, making impossible the subassemblage determination. Assemblage A was identified in three isolates (3/26, 11.5 %), and our results strongly suggest that two isolates belong to subassemblage A2. This study provides information about G. duodenalis genotypes in a rural area of Guinea-Bissau and may contribute for a better understanding of giardiasis epidemiology in this country.

Some pouting caught off the Atlantic coast of Portugal are discarded as unmarketable due to a dark discolouration of the skin and muscle. This study investigates the cause of this condition, describes the new parasite species responsible, and highlights the importance of educating those in charge of premarket inspection of food fish in order to reduce likelihood that consumers will eat infected fish. Macroscopically, infected fish showed considerable heterogeneity in darkening of the skin and hypaxial and epaxial muscles. Microscopical observation revealed bipolar nematode eggs in varying stages of development arranged in a linear pattern along muscle fibers. Histopathology confirmed the presence of eggs of a nematode of the genus Huffmanela Moravec, 1987 as the cause of muscle darkening and established a relationship between infection intensity and consequent darkened appearance of the tissues. The eggs are oval or barrel-shaped, with a smooth surface and polar plugs at opposite ends. The thin outer vitelline membrane is smooth and lacks ornamentation. Under light microscopy, the main eggshell of older eggs exhibits the outermost delicate and smooth vitelline membrane, and a thicker layer, correspondent to chitinous and chondroitin proteoglycan layers. Scanning electron microscopy of eggs confirmed light microscopic studies, namely the presence of a smooth vitelline membrane surrounding the egg. Microscopic and ultrastructural characteristics of eggs, and a new host family in a new geographic area, all suggest that a new species, herein named Huffmanela lusitana sp. n. is involved.