Hemostatic potential of latex proteases from Tabernaemontana divaricata (L.) R. Br. ex. Roem. and Schult. and Artocarpus altilis (Parkinson ex. F.A. Zorn) Forsberg.

Pharmacological properties exhibited by latex of plants are due to various biologically active compounds including several proteolytic enzymes. Present study evaluates hemostatic potential of Tabernaemontana divaricata and Artocarpus altilis from Apocynaceae and Moraceae families respectively. The latex of these plants were initially subjected to dialysis and crude extracts were estimated for proteolytic activity using casein as the substrate. Mean caseinolytic activity for 100 µg of latex protein was found to be 56.16 ± 0.57 and 45 ± 0.3 U/h for T. divaricata and A. altilis respectively. Caseinolytic activity by both the plant extracts was higher than standard proteases, papain and trypsin. However the difference was significant (p < 0.05) with papain alone. Crude enzymes (CE) from both plants exhibited coagulant activity on human platelet poor plasma by recalcification time. A significant reduction in clotting time was exhibited by T. divaricata compared to A. altilis (p < 0.05). These results were further substantiated with fibrinogen agarose plate assay. Crude enzyme of both plants also hydrolyzed blood clot. Mean % of thrombolysis by T. divaricata was 80.75 ± 1.2 and that of A. altilis was 70.24 ± 1.52. Inhibition studies confirmed cysteine protease nature of CE. Comparative analysis revealed T. divaricata to be the best among the two for its hemostatic potential. This study scientifically validates the use of latex from these plants in the management of fresh cuts or wounds.

High expression levels of nuclear factor kappa B and gelatinases in the tumorigenesis of oral squamous cell carcinoma.

BACKGROUND: The cellular mechanisms involved in transformation of a premalignant/potentially malignant oral lesion to a malignant one remain unclear. Previous studies have documented a direct involvement of matrix metalloproteinase (MMP) overexpression in the development and progression of head and neck squamous cell carcinoma (HNSCC). MMP activation, particularly MMP2 and MMP9, observed in different cancers, has been shown to be mediated via the transcription factor nuclear factor kappa B (NF-kappaB). The present study analyzes the clinical significance of gelatinases and NF-kappaB in various histologic phases of human oral tumor progression. METHODS: Methodology included immunohistochemistry for MMP2, MMP9, p50, and p65 components of NF-kappaB and IkappaBalpha (inhibitor kappaBalpha). Gelatin zymography was carried out to determine the extent of gelatinolytic activity. Western blotting was used to confirm the gelatinolytic bands of zymogram, and electrophoretic mobility shift assay (EMSA) was carried out to confirm NF-kappaB activation. RESULTS: A gradual increase was evident in the intensity of the expression and gelatinolytic activity of gelatinase paralleling the histologic progression of malignancy. This finding supports the histologic evidence of tumor invasion occurring in the transition between premalignancy and invasive cancer. Nuclear translocation of NF-kappaB (p50-p65 form) gradually progresses through the premalignant phase of oral tissue to the invasive phase, showing NF-kappaB activation during oral tumorigenesis. NF-kappaB activation correlatively paralleled the pattern of expression of gelatinases. CONCLUSIONS: The results of this study suggest a regulatory role for NF-kappaB on activation of gelatinases during malignant transformation in the oral mucosa.