We established seven hybridomas secreting monoclonal antibodies (MoAbs) against the venom from Bothrops jararaca. Six of them were demonstrated to specifically recognize botrocetin, a venom protein which binds with von Willebrand factor (vWf) and induces platelet agglutination. Two of them, BCT4-3 and BCT115-2 MoAbs, could significantly inhibit botrocetin binding with plasma vWf. BCT4-3 could react slightly with a monolayer of human endothelial cells (ECs), and BCT4-3 binding to ECs was drastically enhanced by the coexistence of human plasma in a dose-dependent manner, indicating that a biological modulator structurally resembling botrocetin is created initially on the EC surface complexed with some plasma proteins. Botrocetin-like components could be immuno-purified only by immobilized BCT4-3, but not by the other immobilized MoAbs, from umbilical vein extracts. Interestingly, the immunoisolated materials were identified to consist essentially of fibronectin (Fn) and a 130 kDa molecule, and this complex bound to vWf in the extracts. Depletion of Fn from plasma decreased BCT4-3 binding to ECs. The epitope of BCT4-3 expressed on the endothelial surface, comprising plasma Fn and the coisolated 130 kDa molecule, is proposed to be a physiological modulator structurally mimicking botrocetin, and essentially supporting vWf-binding to injured endothelium and subsequently to circulating platelets.
Whip scorpions (Thelyphonida), comprising an order in the class Arachnida, are distributed from tropical to temperate zones. Two species occur exclusively in Japan and Taiwan, but the border of their distributional ranges is ambiguous in the Central Ryukyus (Japan). We collected new specimens from the Central Ryukyus and revealed that the border of distribution of the two species lies between the Central and Southern Ryukyus, i.e., the Kerama Gap. Moreover, the estimated divergence time (15.8 Mya) of the two species, based on the mitochondrial COI gene, was older than the recently estimated time (1.55 Mya) of formation of the Kerama Gap. These results highlight the risks of a priori assumption solely on the basis of geological data for applying it as a calibration point to some terrestrial animals in this region. Typopeltis stimpsonii was genetically divided into four lineages, two of which turned out to be endemic to the Okinawa Islands. All specimens from the main island of Japan and Shikoku were in one lineage, which was also found in the Amami Islands and Hachijojima Island. This suggests that these specimens may have been dispersed by human activity. Typopeltis crucifer included five genetic lineages. Species collected from Ishigakijima and Iriomotejima Islands were genetically diversified not between the borders of these islands but within Ishigakijima Island. This study also suggests that phylogenetic diversity of the species in the Southern Ryukyus have increased through two times of invasions from Taiwan.
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