Rapidly labelled ribonucleic acid from rat liver. Studies in the attachment to ribosomes and stimulation of the incorporation of amino acids into polypeptides in cell-free systems

1. Rapidly labelled RNA from rat liver, either as a complex with DNA (m-RNA-DNA) or with ribosomal RNA (m-RNA-RNA) binds to ribosomes in the polysome region. No binding could be demonstrated with ribosomal RNA or native DNA from Bacillus subtilis. 2. With ribosomes from rat liver, Escherichia coli or hepatoma the m-RNA-DNA stimulated incorporation of amino acids with rat-liver ribosomes only, whereas the m-RNA-RNA complex was effective with ribosomes from E. coli or the hepatoma. 3. Polyuridylic acid was effective as messenger RNA with all three ribosomes but much greater stimulation was obtained with ribosomes from E. coli and the hepatoma. 4. The degree of incorporation of phenylalanine with polyuridylic acid and ribosomes from a hepatoma was decreased by about 50% when ribosomal RNA was present.     

Simplified, Accurate Method for Antibiotic Assay of Clinical Specimens

Large glass plates are used for this modified agar-well diffusion assay method, allowing up to 81 replications on a single plate. With a specially designed agar punch, it is possible to prepare the small agar wells very quickly. The saving in serum resulting from fewer replications of standards with the large plates, and the small volume of the agar wells, makes it economically feasible to use pooled human serum for the standard antibiotic solutions. Methods are described for preparing the standard solutions, and for providing controls for the deterioration of standards and unknowns. Procedures for preparing and maintaining the commonly used assay organisms are presented. Serum specimens are tested directly rather than diluting them to a narrow range of antibiotic concentrations. This is possible because of a procedure for calculations that recognizes the curvilinear relationship between zone sizes and antibiotic concentrations. Adaptation of this method to a number of the commonly used antibiotics is described. With this method, it has been possible to test large numbers of clinical specimens in a minimal time, and with accuracy consistently better than 10%.     

Inactivation of cephalothin and cephaloridine by Staphylococcus aureus

Benner, Ernest J. (University of Washington School of Medicine, Seattle), John V. Bennett, Jean L. Brodie, and William M. M. Kirby. Inactivation of cephalothin and cephaloridine by Staphylococcus aureus. J. Bacteriol. 90:1599-1604. 1965.-Marked differences were observed in the susceptibility of penicillinase-producing staphylococci to cephalothin and cephaloridine. All of 100 strains of penicillin G-resistant Staphylococcus aureus, with the use of a large inoculum, were found to be susceptible to 2 mug/ml of cephalothin, whereas only 50% were susceptible to this concentration of cephaloridine, and 15% required 15 mug/ml or more for inhibition. In contrast, penicillin G-sensitive strains were more susceptible to cephaloridine and did not show the marked inoculum effect observed with the cephaloridine-resistant strains. These differences were due to a much greater destruction of cephaloridine than of cephalothin by staphylococcal penicillinase. Cephaloridine-resistant staphyloccoci were stronger penicillinase producers than were susceptible strains, and the resistant strains were found to inactivate cephaloridine by hydrolysis of the beta-lactam ring. In population studies, cephaloridine-resistant cells differed from methicillin-resistant cells in that they decreased in numbers as the drug concentration was increased, and the survivors in higher drug concentrations were no more resistant than was the parent strain. Treatment with acriflavine eliminated resistance of the cells to both penicillin G and cephaloridine. It was concluded that cephaloridine resistance was due to hydrolysis by penicillinase, and that this was related to the pyridine ring substitution in the cephalosporanic acid nucleus.     

Isolation of deoxyribonucleic acid from mammalian tissues

1. DNA has been isolated from different mammalian tissues. The DNA preparations were free from RNA, protein and polysaccharides and have a similar range of sedimentation coefficients (approx. 24s). 2. Protein was removed by a two-stage extraction with a phenol-cresol mixture by using a detergent with 4-aminosalicylate in the first stage and sodium chloride in the second. 3. Polysaccharides remained in solution when DNA was precipitated with 2-butoxyethanol in the presence of 0.5m-sodium chloride and 1.5m-sodium benzoate. 4. Ribosomal RNA was removed by precipitation in the presence of 3m-sodium chloride at 0 degrees , when DNA remained soluble.     

Biochemical and functional characterization of proteoglycans produced by Sl/Sld murine bone marrow stromal cell lines

The Steel anemia of mice results from an inherited defect in the hematopoietic microenvironment. Proteoglycans synthesized by bone marrow stromal cells are an important functional component of the hematopoietic microenvironment in normal animals. It is thus possible that Steel anemia results from a molecular abnormality involving bone marrow stromal proteoglycans. To investigate this possibility, we studied proteoglycan synthesis in three stromal cell lines from Steel anemic (Sl/Sld) animals and two control stromal cell lines, one (+/+2.4) from a non-anemic littermate, and one (GBl/6) from a normal mouse. Proteoglycans were precursor labelled with 35S sulfate and separated by ion exchange HPLC, CsCl density gradient centrifugation, and molecular sieve HPLC. Glycosaminoglycan (GAG) moieties were characterized by molecular sieve HPLC and enzyme sensitivity. There were no consistent differences in total proteoglycan synthesis, proteoglycan heterogeneity, GAG hydrodynamic size, or enzyme sensitivity among the cell lines studied. Growth factor binding to stromal extracellular matrix (ECM) was studied by co-culture of an IL-3-dependent cell line (FDC-P1) with cell-free ECM preparations from an Sl/Sld and a control (GBl/6) stromal cell line, with and without pre-incubation with IL-3. Cell-free ECM preparations from Sl/Sld and control cell lines supported FDC-P1 growth to an approximately equal extent after pre-incubation with IL-3. FDC-P1 growth support by ECM preparations from both cell lines was also observed without IL-3 pre-incubation, although to a lesser extent, suggesting ECM binding of endogenous growth factors synthesized by the stromal cells.     

Evolutionary origin of aminoglycoside phosphotransferase resistance genes

Summary The protein sequences of seven 3-aminoglycoside phosphotransferases falling into the six identified types and three 6-aminoglycoside phosphotransferases were analyzed to give a rooted phylogenetic tree. This tree supports the origin of these groups of enzymes in an ancestor closely related to the actinomycetes, and that horizontal transfer of the resistance genes occurred, possibly via transposons. The implications for genetic engineering of a novel antibiotic are discussed.     

Population structure and responses to disturbance of the basidiomycete Resinicium bicolor

Summary Resinicium bicolor (Alb. & Schw. ex Fr.) Parm. [=Odontia bicolor (Alb. & Schw. ex Fr.) Bres.] is an outcrossing resupinate basidiomycete associated with root and butt rots of trees, but is itself only very weakly pathogenic. The distribution of genets among every spruce stump in a 70-year-old 1250 m² spruce stand was analysed using somatic incompatibility testing. R. bicolor was present on 40% of 8-to 10-year-old stumps. Nineteen genets were found occupying 32 stumps; yielding probabilities of colonisation following establishment by basidiospores of 0.20–0.24 and by mycelial extension or dispersal of 0.16–0.20. The probability of colonisation decreased with increasing distance from a point of establishment. R. bicolor responded to both enrichment and destructive disturbances by the formation of an extensive cord system which enabled it to colonise discontinuously distributed resources and to overgrow fungi adjacent to it in a single resource unit, including Heterobasidion annosum.     

Complete cDNA sequence of bovine alpha 1-antitrypsin.

A cDNA clone coding for the entire bovine alpha 1-antitrypsin molecule has been isolated from a lambda gt11 bovine liver cDNA library using a human alpha 1-antitrypsin cDNA as a probe. The bovine cDNA was sequenced by the dideoxynucleotide chain termination method. Comparison of the translated amino acid sequence of the bovine alpha 1-antitrypsin with those of the human, baboon, sheep, rat and mouse demonstrates the preservation of most of the critical structural determinants. The bovine and the sheep molecules have a sequence homology of 94% and both the molecules contain four cysteine residues; there is only one cysteine in the others.     

Mute Swans in Great Britain: a review,current status and long-term trends

We review the recent history of the Mute Swan in Great Britain and discuss the factors known to be affecting the population. Following a rise in the national population during the 1950s, numbers decreased sharply during the 1960s, and changed relatively little between 1970/71 and 1984/85. However, there has been considerable regional variation in the fortunes of Mute Swan populations during this period, with dramatic declines in some areas. Although several factors were thought to be contributing to such declines, poisoning from the ingestion of lead fishing weights was shown to be the largest single cause of death amongst swans in a number of areas. Voluntary measures to address this problem were initiated in 1982 and culminated in the banning by law of use of lead weights in 1987.Winter counts were used to investigate the current status and distribution of the Mute Swan in Great Britain and to examine long-term regional trends. The maximum total count reached 12600 birds in January 1990, which compares with an average of 9550 for the previous five winters. However, accounting for birds missed, the population may now number at least 25 000. Peak total numbers have mostly occurred in September, after which numbers remain approximately stable until December and then decline. Patterns of seasonal abundance vary between regions and habitats and these are discussed.The British population has increased dramatically since 1986/87 and reached its highest level for 27 years in 1987/88. There have been recent increases in most regions with record levels being reached mostly in 1987 or 1988, and there has been growth in the numbers on all habitat types, especially on reservoirs, gravel extraction pits and freshwater marshes. The timing of these increases corresponds very closely with the introduction of legislation against the use of lead fishing weights, and the incidence of lead poisoning is known to have been considerably reduced by such measures.