Tissue culture of Ecklonia radiata (Phaeophyceae,Laminariales): effects on growth of light,organic carbon source and vitamins

The effects of light quality and irradiance, and supply of organic carbon and vitamins on the growth of two forms of Ecklonia radiata in tissue culture were examined. A callus of unpigmented cells developed over the cut surface of newly excised explants of stipe. This growth was best in the dark but stopped after 10 weeks. Pigmented, mainly filamentous clumps of cells developed from explants after several weeks in culture. These required light for growth, with growth being enhanced by increasing photon flux density up to 30 μmol photon m^-2 s^-1, with the active spectral component being red light (> 600 nm). The addition to the medium of a range of organic carbon sources or vitamins did not stimulate growth of either culture type in the dark. author for correspondence     

A novel anthelmintic model utilizing jirds, Meriones unguiculatus, infected with Haemonchus contortus

Currently, no in vivo laboratory model is available for evaluating anthelmintics against the important ruminant helminth Haemonchus contortus. This report outlines a novel anthelmintic assay utilizing immunosuppressed (0.02% hydrocortisone in feed) jirds, Meriones unguiculatus, infected with H. contortus. Immunosuppressed jirds were inoculated with approximately 1,000 exsheathed infective larvae of H. contortus, treated per os on day 10 postinoculation (PI), and necropsied on day 13 PI. Each stomach was removed, opened longitudinally, incubated in distilled water at 37 C for 5 hr, fixed in formaldehyde solution, and stored for subsequent examination. Stomach contents were examined using a stereomicroscope (15-45x). A variety of standard anthelmintics has been evaluated in the model; modern broad-spectrum ruminant anthelmintics (benzimidazoles, febantel, ivermectin, levamisole hydrochloride, and milbemycin D) are active uniformly and in most cases at doses (mg/kg) comparable to those required for efficacy against H. contortus in ruminants. This model provides an important new tool to assess preliminarily the activity of experimental drugs against H. contortus in vivo prior to studies in ruminants and also may provide a useful tool for studying host-parasite interactions for H. contortus.     

Lipoxygenase inhibitors enhance the proliferation of human B cells

The addition of drugs which inhibit the lipoxygenase pathways of arachidonic acid metabolism to 5 day cultures of mitogen-stimulated human B cells enhanced the proliferative response more than 10-fold. Several chemically dissimilar lipoxygenase inhibitors increased proliferation in this system, whereas the specific cyclooxygenase inhibitor indomethacin had no effect. A lipoxygenase inhibitor could be added as late as 48 to 72 h after the initiation of culture and still cause a significant increase in B cell proliferation. These drugs increased the proliferation of both peripheral blood B cells and tonsillar B cells activated by Staphylococcus aureus Cowan I or anti-Ig M antibodies, in combination with a crude T cell supernate, a commercial B cell growth factor preparation, or recombinant lymphotoxin. A similar effect was observed in tonsillar B cells purified by counterflow centrifugal elutriation to remove esterase positive accessory cells, suggesting this is a direct effect on the B cell. Lipoxygenase blockade also caused a greater than twofold increase in polyclonal Ig production. The enhanced proliferation caused by lipoxygenase blockade could not be reversed by adding back exogenous leukotrienes or hydroxyeicosatetraenoic acids to the cultures. Furthermore, B cells prelabeled with [3H]arachidonic acid did not produce radiolabeled lipoxygenase metabolites of arachidonic acid under the same culture conditions in which the addition of lipoxygenase inhibitors had a profound effect on proliferation. Thus, lipoxygenase inhibitors markedly stimulate B cell proliferation under a variety of experimental conditions, although the mechanism responsible for this action has not yet been elucidated.     

β-II conformation of all-β proteins can be distinguished from unordered form by circular dichroism

The CD spectrum of certain all-β globular proteins resembles that of unfolded proteins with a characteristic negative band around 200 nm. The conformation of this class is tentatively termed β-II, which had two features that were absent for unfolded proteins. First, β-II proteins usually had CD bands due to aromatic side groups in the near-ultraviolet region. Second, the CD intensities both in the far- and in the near-uv region of these compact and rigid proteins usually showed a sharp transition upon thermal denaturation, whereas those of an unordered form changed linearly with rising temperature.     

Review: Hydrogels for cell immobilization

Hydrogels are being investigated for mammalian cell immobilization. Their material properties can be engineered for biocompatibility, selective permeability, mechanical and chemical stability, and other requirements as specified by the application including uniform cell distribution and a given membrane thickness or mechanical strength. These aqueous gels are attractive for analytical and tissue engineering applications and can be used with immobilization in therapies for various diseases as well as to generate bioartificial organs. Recent advances have broadened the use of hydrogel cell immobilization in biomedical fields. To provide an overview of available technology, this review surveys the current developments in immobilization of mammalian cells in hydrogels. Discussions cover hydrogel requirements for use in adhesion, matrix entrapment, and microencapsulation, the respective processing methods, as well as current applications. (c) 1996 John Wiley & Sons, Inc.     

Reaction mechanism of isomaltooligosaccharides synthesis by α-glucosidase fromAspergillus carbonarious

Summary An -glucosidase fromAspergillus carbonarious CCRC 30414 was employed for investigating the enzymatic synthesis of isomaltooligosaccharides from maltose. The enzyme transferred a glucose unit from the nonreducing end of maltose and other -linked glucosyl oligosaccharides to glucose and other glucosyl oligosaccharides which function as accepting co-substrates. The transfer of a glucose unit occurs most frequently to the 6-OH position of the nonreducing end of acceptor, but transfer to 4-OH position also occurs. Treatment of 30 % (w/v) maltose with the enzyme under optimum conditions afforded more than 50% isomaltooligosaccharides.     

Clonal growth in woody plants: A review

Bulkiness, longevity and solidity of the body in woody plants enable the successive development of accessory shoots and adventitious roots in (1) both proximal and distal positions on organs, (2) both the above-ground and below-ground space, (3) both the aerial and soil environments. In monocotyledons, woody rhizomes play an essential role in the basic growth habit and architectural models. In dicotyledonous and a few gymnospermous trees, attached and successively disconnected ramets play a multilateral role in the pertinent life strategies. The majority of sprouts, coppice shoots and root suckers behave as opportunistic organs (a) serving as means of vegetative reproduction, (b) securing colonization of unoccupied ground, (c) increasing competitive power of the species within the community, (d) increasing survival rate of the stressed/disturbed genet in marginal habitats, (e) forming replacement for ageing or damaged organs, and (f) enabling reiteration of the genet's entire architectural model.     

The distribution of colony-forming cells in the kidney

Abstract. A method is described for producing outgrowths of small nephron segments (average 24 cells) in culture. The method was used to estimate an overall colony-forming efficiency of 4.6% for cells constituting the segments. Efficiency was found to be lower for thick segments (1%) than for thin segments (6%) from Henle's loop. The latter higher level indicates that precursor cells are concentrated near the middle of the nephron. For comparison, a two-dose irradiation technique was used to calculate a mean number of 5 ± 2 (SE) clonogens per segment producing outgrowths. This tended to be higher than the value of about 1 calculated from the 65% of segments producing outgrowths, as expected if the remaining segments contained no clonogens.     

Characterization of phospholipase A2 activation by plasmin in cultured bovine endothelial cells

Treatment of cultured bovine carotid artery endothelial cells with 0.1 µM human plasmin has been reported to induce a receptor-mediated short burst of arachidonate release, which is a pertussis toxin-sensitive and extracellular calcium-dependent reaction. Plasmin-induced calcium influx in cells was significantly inhibited by pretreatment with pertussis toxin, indicating that the former was coupled with a pertussis toxin-sensitive guanosine 5-triphosphate (GTP)-binding protein. Plasmin significantly induced the formation of lysophosphatidylcholine but not lysophosphatidylethanolamine. A cellular phospholipase A₂ with an arachidonyl specificity at the sn-2 position of phosphatidylcholine, which required submicromolar calcium, was identified as a cytosolic phospholipase A₂ by immunoblot analysis. By a cell-free enzyme activity assay and immunoblot analysis, plasmin was found to induce a translocation of the cytosolic phospholipase A₂ from the cytosol to the membrane. Taken together, the results suggest that plasmin bound to its putative receptor and activated a GTP-binding protein coupled to calcium influx channel, followed by translocation and activation of cytosolic phospholipase A₂ in endothelial cells.