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In vivo and in vitro evaluation for immunomodulatory activity of three marine animal extracts with reference to phagocytosis

Sun, 01 Dec 2002 00:00:00 GMT

Title: In vivo and in vitro evaluation for immunomodulatory activity of three marine animal extracts with reference to phagocytosis Authors: Ponkshe, C A; Indap, Madhavi M Abstract: The whole body ether extracts of a marine prawn Nematopaleamon tenuipes and two gastropods viz. Euchelus asper and Hemifusus pugilinus, obtained by Soxhlet extraction and cold percolation were tested for their effects on phagocytosis by in vitro (slide method) and by in vivo (carbon clearance) methods. Extract of E. asper exhibits immunostimulatory activity in vivo and immunosuppressant activity in vivo. The in vitro test for N. tenuipes and H. pugilinus shows biphasic activity, but the former shows immunostimulatory while the later shows immunosuppressant activity in vivo test. Page(s): 1399-1402

Purification and some important characters of extracellular inulinase of Alternaria alternata (Fr.) Keissler

Sun, 01 Dec 2002 00:00:00 GMT

Title: Purification and some important characters of extracellular inulinase of Alternaria alternata (Fr.) Keissler Authors: Hamdy, Hossam S Abstract: Protein prceipitate of cell - free dialysate of extracellular inulinase (2,1-β-fructan fructanohydrolase, EC 3.2.1.7) of A. alternata was maximally obtained by methanol. Such protein was fractionated by using 2-step column chromatography on Sephadex G₁₅₀ and DEAE-cellulose. The partially purified enzyme had activity of 81 x 10³ U/mg protein, with a yield of 69% of the original activity and the fold of purification was 62. Optimum temperature and pH for the activity of the purified enzyme were found to be 55°C and 4.5, respectively. The enzyme was found to be stable up to 55°C and in pH range of 4 to 5. Ba²⁺ and Ca²⁺ were found to stimulate the enzyme activity while Cu²⁺, Fc³⁺, Hg²⁺, and iodoaectate were recorded as strong inhibitors. T_1/2 of the enzyme was estimated to be two weeks and its apparent K_m was calculated to be 0.066 M. The enzyme recorded hydrolyzing activity against sucrose and raffinose recording I/S ratio of 0.50. Molecular mass of the enzyme preparation was estimated by gel filtration and found to be 115 ± 5 kDa. Page(s): 1393-1398

Evaluation of parameters for high efficiency gene transfer via particle bombardment in Indian mulberry

Sun, 01 Dec 2002 00:00:00 GMT

Title: Evaluation of parameters for high efficiency gene transfer via particle bombardment in Indian mulberry Authors: Bhatnagar, Somika; Kapur, Anita; Khurana, Paramjit Abstract: Particle bombardment is a popular method of direct gene delivery into cell, tissue and organs since it requires minimum pre- and post-bombardment manipulation. In addition, this technique is much easier and fast to perform with intact tissue/ organ and reduces the period of in vitro culture. Genetic transformation of mulberry, Mortis indica cv. K2 was attempted by particle bombardment using hypocotyl, cotyledon, leaf and leaf callus explants. The effect of various physical and biological parameters during bombardment were studied by the histochemical localization of GUS reporter gene following two days of bombardment and by assessing the number of blue spots per explant. p35SGUSINT was used for optimization of different parameters. The percentage of GUS positive ex plants was very low with tungsten (20%) as compared to gold particles (36%) indicating tungsten toxicity to the tissue. Maximum GUS activity was observed at 1100 psi helium pressure and 9 cm target distance for hypocotyl, cotyledon and leaf. Double bombardment of explants with 10 µ g of DNA loaded on macrocarriers clearly yielded a better (up to 56%) result as compared to a single bombardment (30%). Amongst the various plasmids tested, pBI221 gave the highest (100%) GUS positive explants in the leaf callus. Page(s): 1387-1392

In vitro regeneration of roots of Phyla nodiflora and Leptadenia reticulata, and comparison of roots from cultured and natural plants for secondary metabolites

Sun, 01 Dec 2002 00:00:00 GMT

Title: In vitro regeneration of roots of Phyla nodiflora and Leptadenia reticulata, and comparison of roots from cultured and natural plants for secondary metabolites Authors: Bhatt, Trupti; Jain, Vandana; Jayathirtha, M G; Banerjee, G; Mishra, S H Abstract: Adventitious roots, generated using leaf explants of P. nodiflora, and meristem explants of L. reticulate, were cultured on Murashige and Skoog (MS) medium supplemented with napthylacetic acid (2 µM) and indole butyric acid (3µM) respectively. After 30 days, subculturing of roots in liquid MS medium with napthylacetic acid (1.5 µM) for P.nodiflora and indole butyric acid (3 µM) for L. reticulata afforded considerable increase in root mass. HPTLC profiles and microscopic examination of transverse sections of ill vitro and naturally grown roots provided information on secondary metabolite accumulation vis-à-vis developmental stages of the root. Page(s): 1382-1386