Effect of anticoagulants on the plasma hyaluronidase activities.

Mammalian hyaluronidases (HAases) are an endo-beta-N-acetyl-hexosaminidases that degrade hyaluronan (HA) and have been implicated in diverse pathophysiological functions. Several pathological conditions, such as diabetes, monoclonal gammapathy, and bladder and prostate tumors, report the distorted plasma HAase activity. However, the plasma HAase (hHyal-1) activity has been presumed to change with the circulating HA level and serves as an early marker for several diseases. It has been generally practised to use the anticoagulants such as tri-sodium citrate/di-sodium EDTA/heparin for the preparation of plasma for both biochemical and clinical analyses. In the present investigation, the effect of anticoagulants on plasma HAaseactivity was evaluated and compared with the serum HAase activity that is devoid of anticoagulants as no study provides information in this regard. The results suggested that the plasma HAase activity in the presence of the recommended concentration of EDTA was highly comparable/similar to that of the serum HAase activity. In contrast, citrated or heparinized plasma recorded a significantly reduced level of activity than that of the serum HAase activity. In conclusion, our results suggested that the EDTA-treated plasma samples are a better choice compared with heparin and citrated samples to assess the HAase activity.

Antimicrobial properties of a non-toxic glycoprotein (WSG) from Withania somnifera (Ashwagandha).

A monomeric glycoprotein with a molecular mass of 28 kDa in SDS-PAGE was isolated from the Withania somnifera root tubers. The protein designated WSG (Withania somnifera glycoprotein) demonstrated potent antimicrobial activity against the phytopathogenic fungi and bacteria tested. Antifungal effect has been demonstrated in that WSG exerts a fungistastic effect by inhibiting spore germination and hyphal growth in the tested fungi. WSG showed potent antifungal activity against Aspergillus flavus, Fusarium oxysporum, F. verticilloides and antibacterial activity against Clvibacter michiganensis subsp. michiganensis. WSG is an acidic, non-toxic (trypsin-chymotrypsin) protease inhibitor. These results encourage further studies of WSG as a potential therapeutic agent for its antifungal activity.

The anti-snake venom properties of Tamarindus indica (leguminosae) seed extract.

In Indian traditional medicine, various plants have been used widely as a remedy for treating snake bites. The aim of this study was to evaluate the effect of Tamarindus indica seed extract on the pharmacological as well as the enzymatic effects induced by V. russelli venom. Tamarind seed extract inhibited the PLA(2), protease, hyaluronidase, l-amino acid oxidase and 5'-nucleotidase enzyme activities of venom in a dose-dependent manner. These are the major hydrolytic enzymes responsible for the early effects of envenomation, such as local tissue damage, inflammation and hypotension. Furthermore, the extract neutralized the degradation of the Bbeta chain of human fibrinogen and indirect hemolysis caused by venom. It was also observed that the extract exerted a moderate effect on the clotting time, prolonging it only to a small extent. Edema, hemorrhage and myotoxic effects including lethality, induced by venom were neutralized significantly when different doses of the extract were preincubated with venom before the assays. On the other hand, animals that received extract 10 min after the injection of venom were protected from venom induced toxicity. Since it inhibits hydrolytic enzymes and pharmacological effects, it may be used as an alternative treatment to serum therapy and, in addition, as a rich source of potential inhibitors of PLA(2), metalloproteinases, serine proteases, hyaluronidases and 5 cent-nucleotidases, the enzymes involved in several physiopathological human and animal diseases.