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The signals and the molecular machinery mediating release of dense matrix granules from pathogenic protozoan parasites are unknown. We compared the secretion of the endogenous dense granule marker GRA3 in Toxoplasma gondii with the release of a stably transfected foreign reporter, ␤-lactamase, that localizes to parasite dense granules. Both proteins were released constitutively in a calcium-independent fashion, as shown using both intact and streptolysin O-permeabilized parasites. N-Ethylmaleimide and recombinant bovine Rab-guanine dissociation inhibitor inhibited ␤-lactamase secretion in permeabilized parasites, whereas recombinant hamster N-ethylmaleimide-sensitive fusion protein and bovine ␣-SNAP augmented release. Guanosine 5-3-O-(thio)triphosphate, but not cAMP, augmented secretion in the presence but not in the absence of ATP. The T. gondii NSF/SNAP/SNARE/Rab machinery participates in dense granule release using parasite protein components that can interact functionally with their mammalian homologues.

No abstract

A refugium is generally understood as an area where temperate species survive cold periods, such as the Iberian, Italian, or Balkan Peninsulas in Europe. Strictly speaking, this definition refers to what is known as a glacial refugium. However, there are various types of lesser-known refugia such as the interglacial refugium, which denotes a mountainous region at low latitudes, such as the Pyrenees, where species adapted to the cold survive during interstadial periods. The small-vertebrate association from the sequence of Cova Colomera, which is located on the southern face of the Pyrenees and contains the final cold spell of the Late Pleistocene and the beginnings of the temperate period in which we currently find ourselves (the Holocene), could constitute the first fossil evidence of such an interglacial refugium, thus providing new paleoecological data on the phenomenon.