Monomeric midkine induces tumor cell proliferation in the absence of cell-surface proteoglycan binding

Midkine (MK), a retinoic acid-inducible heparin-binding protein, is a mitogen which initiates a cascade of intracellular protein tyrosine phosphorylation mediated by the JAK/STAT pathway after binding to its high affinity p200(+)/MKR cell surface receptor in the G401 cell line [Ratovitski, E. A. (1998) J. Biol. Chem. 273, 3654-3660]. In this study, we determined the biophysical characteristics of purified recombinant murine MK and analyzed the requirements for ligand multimerization and cell surface proteoglycan binding for the G401 cell mitogenic activity of MK. Our studies indicate that the secreted form of MK (M = 13 kDa) exists in solution as an asymmetric monomer with a frictional coefficient of 1. 48 and a Stokes radius of 23.7 A. By constructing bead models of MK using the program AtoB and the program HYDRO to predict the hydrodynamic properties of each model, our data suggest that MK has a dumb-bell shape in solution composed of independent N- and C-terminal domains separated by an extended linker. This asymmetric MK monomer is a biologically active ligand with mitogenic activity on G401 cells in vitro. Neither heparin-induced formation of noncovalent MK multimers nor tissue transglutaminase II covalent multimerization of MK enhanced MK mitogenic activity in this system. Since neither heparin competition nor cell treatment with chondroitinase ABC or heparinase III abolished the mitogenic effects of MK on G401 cells, cell-surface proteoglycan binding by MK does not appear to be a requirement for its observed mitogenic effects. These results provide strong evidence that the MK-specific p200(+)/MKR has distinctive biochemical properties which distinguish it from the receptor tyrosine phosphatase cell-surface proteoglycan PTPzeta/RPTPbeta and support the hypothesis that the diverse biological effects of MK are mediated by multiple cell-specific signal transduction receptors.     

Anthracyclines: isolation of overproducing strains by the selection and genetic recombination of putative regulatory mutants of Streptomycespeucetius var. caesius

In Streptomyces peucetius var. caesius, the production of anthracyclines was suppressed either by 330 mM d-glucose or 25 mM phosphate. In addition, the anthracycline doxorubicin and the glucose analogue 2-deoxyglucose inhibited the growth of this microorganism at concentrations of 0.025 mM and 10 mM respectively. Spontaneous and induced mutants, resistant to the action of these compounds, were isolated, tested and chosen by their ability to overproduce anthracyclines. Genetic recombination between representative mutants was carried out by the protoplast fusion technique. Some recombinants carrying resistance to doxorubicin, phosphate and 2-deoxyglucose produced more than 40-fold greater levels of anthracyclines than those obtained with the parental strain. This improvement resulted in total antibiotic titres of more than 2 g/l culture medium at 6 days of fermentation. Received: 14 April 1997 / Received revision: 19 June 1997 / Accepted: 4 July 1997     

Deposition of sediments during a flood event on seasonally flooded forests of the lower Orinoco River and two of its black-water tributaries, Venezuela

Deposition of suspended sediments andtheir associated nutrients were estimated during theflood event of 1995 in the seasonally flooded forestsof the Mapire and Caura Rivers, two black-watertributaries of lower Orinoco, and on two islands ofthis white-water river. The deposition spanned a widerange from 0.07 kg m^–2 in the depositional barforests of the Mapire River to 73.60 kg m^–2 onthe flooded forests of the Orinoco Island site calledJarizo. This variation is associated with the dynamicnature of sediment mobilization, transport anddeposition, as well as with the different geomorphicenvironments and erosion processes upstream from thestudy sites. The deposited sediment in all the studyareas was highly quartzitic with a relatively highcontent of kaolinite and goethite. Only in thesediment of the Orinoco Islands was mica (illite)identified in a relatively high proportion. Thesemineralogical results reflect the intense weatheringprocesses in the catchment areas of the study rivers.The chemical composition of the deposited sedimentsshowed a great variability among the different studyareas, which is in part related to the mineralcomposition of the sediments and their particle sizedistribution. The highest concentrations of K, Caand Mg were found on the clay sediments of the Orinocoagricultural island. The total amount of depositednutrients varied over a wide range, which isinfluenced by the amount of deposited sediments. Inthe Jarizo Island site of lower Orinoco were depositedthe largest amount of nutrients. In the floodedforests of the Mapire River, the nutrient contributionby the deposited sediments to the nutrient cycling isrelatively low in the depositional-bar forests andpractically nonexistent in the forests sites onterraces.