RESUMO
Digestive carbohydrases are present in many species of hematophagous Arthropoda, including ticks. In this work, Amblyomma cajennense (Ixodidae) midgut digestive carbohydrases were tracked with different substrates, resulting in the identification of a chitinase and an N-acetyl-ß-glucosaminidase and the first description of a digestive α-L-fucosidase in ticks. α-L-fucosidases are involved in various physiological processes, and digestive α-L-fucosidases have been shown to be present in other types of organisms. Amblyomma cajennense α-L-fucosidase activity was isolated using acidic and salting-out precipitations and chromatographic steps in hydrophobic and cation-exchange columns. The specificity of the isolated activity as an α-L-fucosidase was confirmed by the hydrolysis of 4-methylumbelliferyl α-L-fucopyranoside and the natural substrate fucoidan and the inhibition by fucose and deoxyfuconojirimycin. The isolated activity of α-L-fucosidase forms oligomers with molecular mass of 140 kDa or 150 kDa as determined by gel filtration and non-reducing SDS-PAGE, respectively. This particular fucosidase has an optimum pH of 5.3, is stable even at high temperatures (stable for at least 2h at 50 °C), has a Km of 45 µM to the substrate 4-methylumbelliferyl α-L-fucopyranoside and IC 50% of 327 µM to fucose and 42 pM to deoxyfuconojirimycin. The presence of digestive fucosidases in hematophagous Arthropoda may be related to defence mechanisms against host-parasite interactions.
Assuntos
Acetilglucosaminidase/metabolismo , Metabolismo dos Carboidratos , Quitinases/metabolismo , Carrapatos/metabolismo , alfa-L-Fucosidase/metabolismo , Animais , Digestão , Feminino , Coelhos , alfa-L-Fucosidase/isolamento & purificaçãoRESUMO
To study whether treatment with heparin (HEP) attenuates intestinal dysfunction caused by ischemia (I) and reperfusion (R), rats were treated with HEP (100 U/kg intravenously) or saline solution (SS) before I (60 min), which was produced by occlusion of the superior mesenteric artery, and R (120 min). After I or I/R, we mounted 2-cm jejunal segment in an organ bath to study neurogenic contractions stimulated by electrical pulses or KCl, using a digital recording system. Thin jejunal slices were stained with hematoxylin and eosin for optical microscopy. Compared with the sham group, jejunal contractions were similar in the I + HEP and the I/R + HEP groups, but reduced in the I + SS and the I/R + SS groups. The jejunal enteric nerves were damaged in the I + SS and the I/R + SS, but not in the I + HEP and the I/R + HEP cohorts. These results suggested that HEP attenuated intestinal dysfunction caused by I and I/R.