Determination Of Hyaluronidase Activity Research Articles

Determination of hyaluronidase activity in Tityus spp. Scorpion venoms and its inhibition by Brazilian antivenoms

Hyaluronidases (HYALs) are enzymes ubiquitously found in venoms from diverse animals and seem to be related to venom spreading. HYAL activity might be important to Tityus spp. envenoming, since anti-Tityus serrulatus HYAL (TsHYAL) rabbit antibodies neutralize T. serrulatus venom (TsV) lethality. The present work aimed to verify and compare HYAL activity of venoms from other Brazilian Tityus spp. (Tityus bahiensis, Tityus stigmurus and Tityus obscurus) and to test whether anti-TsHYAL antibodies and Brazilian horse therapeutic scorpion antivenom (produced by Fundação Ezequiel Dias (FUNED), Butantan and Vital Brazil Institutes) can recognize and inhibit HYAL activity from these venoms. In ELISA assays, anti-TsHYAL and scorpion antivenoms recognized T. serrulatus, T. bahiensis and T. stigmurus venoms, however, they demonstrated weaker reaction with T. obscurus, which was also observed in Western blotting assay. Epitope mapping by SPOT assay revealed different binding patterns for each antivenom. The assay showed a weaker binding of scorpion antivenom produced by FUNED to peptides recognized by anti-TsHYAL antibodies. Anti-TsHYAL antibodies and antivenoms produced by Butantan and Vital Brazil institutes inhibited HYAL activity of all tested venoms in vitro, whereas FUNED antivenom did not show the same property. These results call attention to the importance of hyaluronidase inhibition, that can aid the improvement of antivenom production.

Untersuchungen zur hyaluronidase-aktivität β-hämolysierender Streptokokken der Lancefield-Gruppe C

A total of 110 strains of beta-hemolytic streptococci, belonging to serogroup C (Lancefield), isolated from horses (71 S. zooepidemicus, 27 S. equisimilis and 12 S. equi) as well as 5 reference strains were tested for their ability to produce hyaluronidase. The determinations were carried out in a culture test on agarose gel and in a liquid test system (turbidity test according to DiFerrante). The results of both methods used showed that the three Streptococcus species could be differentiated by the relative quantitative determination of hyaluronidase activity. S. equisimilis strains produce 5 to 10 times more hyaluronidase than those of S. zooepidemicus. The strains of S. equi did not show enzyme production. In addition parallel tests by determination of final N-acetyl-D-glucosamine groups and by turbidity test were done to confirm the hyaluronidase activity of S. equisimilis strains. The comparison of results (Fig. 3) obtained from these tests, showed a good correspondence as demonstrated by a correlation index of 0,93.

Identification of tissue and bacterial hyaluronidase in human saliva.

Determination of hyaluronidase activity in the saliva of human subjects has shown that two types of hyaluronidase, distinguishable on the basis of their different pH optima, are present. One is considered to be of bacterial origina and the other of tissue origin.

Determination of hyaluronidase activity in dental plaque material

Hyaluronidase activity in dental plaque material was studied using hyaluronic acid as substrate and a viscosimetric method for determining the activity. Although the enzyme activity varied between different patients, two optima at pH 5.5 and 6.5 were found, suggesting the presence of more than one enzyme. A marked decrease in enzyme activity was found even after 1 day storage at ‐20°C. Gel filtration experiments on Sephadex G‐200 revealed at least 2 different fractions, one in the void volume and one close to the void volume.