Isolation of rat hepatocyte plasma membranes. I. Presence of the three major domains

A rat liver plasma membrane preparation was isolated and characterized both biochemically and morphologically. The isolation procedure was rapid, simple and effective in producing a membrane fraction with the following biochemical characteristics: approximately 40-fold enrichment in three plasma membrane markers, 5'-nucleotidase, alkaline phosphodiesterase I (both putative bile canalicular membrane enzymes), and the asialo-glycoprotein (ASGP) receptor (a membrane glycoprotein present along the sinusoidal front of hepatocytes); a yield of each of these plasma membrane markers that averaged approximately 16%; and minimal contamination by lysosomes, nuclei, and mitochondria, but persistent contamination by elements of the endoplasmic reticulum. Morphological analysis of the preparation revealed that all three major domains of the hepatocyte plasma membrane (sinusoidal, lateral, and bile canalicular) were present in substantial amounts. The identification of sinusoidal membrane was further confirmed when ASGP binding sites were localized predominantly to this membrane in the isolated PM using electron microscope autoradiography. By morphometry, the sinusoidal front membrane accounted for 47% of the total membrane in the preparation, whereas the lateral surface and bile canalicular membrane accounted for 6.8% and 23% respectively. This is the first report of such a large fraction of sinusoidal membrane in a liver plasma membrane preparation.     

Cloned nodulation genes of Rhizobium leguminosarum determine host-range specificity

Summary Three nodulation-deficient (nod) mutants of Rhizobium leguminosarum were isolated following insertion of the transposon Tn5 into pRL1JI, the R. leguminosarum plasmid known to carry the nodulation genes. DNA adjacent to the nod: Tn5 alleles was subcloned and used to probe a cosmid clone bank containing DNA from a Rhizobium strain carrying pRL1JI. Two cosmid clones which showed homology with the probe contained about 10 kb of DNA in common. The R. leguminosarum host-range determinants were found to be present within this 10 kb common region since either of the cosmid clones could enable a cured R. phaseoli strain to nodulate peas instead of Phaseolus beans, its normal host. Electron microscopy of nodules induced by Rhizobium strains cured of their normal symbiotic plasmid but containing either of the two cosmid clones showed bacteroid-forms surrounded by a peri-bacteroid membrane, indicating that normal infection had occurred. Thus it is clear that this 10 kb region of nodDNA carries the genes that determine host range and that relatively few bacterial genes may be involved in nodule and bacteroid development.     

Annotated chromosome counts of czechoslovak plants (31–60) (Materials for “Flóra ČSSR”—3)

Chromosome counts of the following 30 taxa (106 populations) are given:Betonica officinalis (2n=16);Bidens frondosus (2n=48);Calamagrostis arundinacea (2n=28+0–2B);Dianthus carthusianorum subsp.latifolius (2n=30);Festuca gigantea (2n=42, 42+2B);Hypericum perforatum (2n=32);Koeleria macrantha (2n=28);Kohlrauschia prolifera (2n=30);Lilium martagon (2n=24+0–2B);Melica ciliata (2n=18);Poa remota (2n=14);Ranunculus polyanthemos (2n=16);R. sardous subsp.sardous (2n=16);Roegneria canina (2n=28+0–1B);Rudbeckia laciniata (2n=76);Scabiosa canescens (2n=16);Serratula tinctoria (2n=22);Seseli elatum subsp.heterophyllum var.beckii (2n=18);S. hippomarathrum (2n=20);Thlaspicaerulescens caerulescens subsp.tatrense (2n=14);Trifolium alpestre (2n=16);T. avense (2n=14);T. medium (2n=79, 80+0–2B, 82);T. rubens (2n=16);Veronica officinalis subsp. alpestris (2n=36);Vincetoxicum hirundinaria (2n=22);Vulpia bromoides (2n=14);Zerna benekenii (2n=28)Z. monoclada (2n=28+0–8B);Z. ramosa (2n=42). Remarks on taxonomy, nomenclature and chorology for some of these taxa are given.     

Preparative Electrophoresis of Isotopically Labeled L-Cell Interferons

The preparative method of polyacrylamide gel electrophoresis was adapted for purification and characterization of isotopically labeled L-cell interferons. Re-covery of interferon activity was quantitative, and purification and resolution were comparable to those obtained by analytical polyacrylamide gel electrophoresis. Ultimate specific activities attainable ranged from 2 x 10(6) to 3 x 10(6) international units per mg of protein.     

Effects of the new antifungal antibiotic mucidin

The antibiotic activity of the antifungal substance mucidin was compared with the activity of nystatin and pimaricin. The antibiotics were tested by the plate method using 19 fungal species, mainly phytopathogenic ones. Toward 14 species, mucidin was ten times more active than nystatin and pimaricin, toward 5 species the activities were roughly the same. The antibiotics differed also in the sharpness of the inhibition zone boundaries.