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Biomedical subjects

A Rosner

Publications and source records attributed to A Rosner.

At least 37 records · Page 2Linked to original sources

The effects of position cues on the appearance of stimulus elements in a bistable apparent movement display.

A modified version of the Ternus display was used to assess the relative effects of element position cues on reports of group and end-to-end movement. In this display, two rows of stimulus elements are joined by connecting lines. In one version of the display, the connecting lines remain stationary across frames, facilitating the interpretation that the associated stimulus elements also remain stationary. In another version of the display, one end of the connecting lines shifts horizontally from frame to fame, facilitating the interpretation that the associated stimulus elements have also shifted. The experiment showed that when the connecting lines remain stationary, reports of end-to-end movement increase, regardless of the interstimulus interval (ISI) at which the frames alternate. When the connecting lines shift, reports of group movement increase, regardless of ISI. Theoretical interpretations of the results involving both relatively low-level motion signals and higher order perceptual influences are considered.

Attention↗

Nucleotide sequence of the capsid protein gene of potato virus Y (PVY).

The nucleotide sequence of the 3' terminal region of potato virus Y (PVY) was determined. Starting with a poly(A) tail of 18 residues a non-coding region of 335 nucleotides precedes the region encoding for the virus coat protein (cp) 801 nucleotides long ending with a TGA. This region was located by comparing the predicted amino acid sequence with the one determined for the PVY capsid protein by Shukla et al. (1). Both sequences contained 267 amino acids sharing about 94% homology. They differ, however, at several positions presumably due to base transitions within their respective nucleotide sequences. Restriction endonuclease sites in and around the cp coding region were identified.

Base Sequence↗

A simple procedure for the extraction of double-stranded RNA from virus-infected plants.

A simple procedure for the isolation of double-stranded (ds) RNA from virus-infected plants is described. The method is based on grinding plant tissue in 4% p-aminosalicylic acid and recovery of ds RNA by phenol extraction and precipitation with 30% ethanol. The presence of both negative and positive virus RNA strands in RNA fractionated in agarose gels was verified by Northern blot hybridization with polynucleotide kinase labelled genomic RNA or complementary DNA (cDNA) probes. The procedure enabled detection of three major ds RNA species (MWs 4.2, 1.05 and 0.48 X 10(6)) and at least 4 minor bands with estimated MWs of 3.5, 2.5, 2.2 and 2.0 X 10(6) in Nicotiana tabacum plants systemically infected with tobacco mosaic virus (TMV). Cucumber mosaic virus (CMV)-infected Pachystachys coccinea plants contained 2 minor bands of MWs 0.49 and 0.35 X 10(6) in addition to the previously described 4 major ds RNAs and ds CARNA 5 (MW 0.22 X 10(6)). The patterns of ds RNA are useful for diagnosing natural infections of CMV and TMV in N. glauca plants and of citrus tristeza virus in Citrus spp.

Mosaic Viruses↗

Evaluation of immunological methods for detection of bovine growth hormone (BGH) produced in E. coli.

The use of several immunological methods for studies on synthesis of bovine growth hormone (BGH) by E. coli is described here. The ELISA procedure was shown to be the least sensitive and unfit for assaying BGH in E. coli extracts. The solid-phase radioimmunoassay (RIA) proved to be highly sensitive, but since E. coli extract itself (not containing BGH) interfered with the immunological reaction, its use for measuring BGH was practically limited. The best adequate procedure proved to be radioimmunoassay in solution, which was not adversely affected by the E. coli extract and was sufficiently sensitive to detect nanogram quantities of BGH. The size of the BGH produced by normal bacterial cells was investigated by protein fractionation, transfer to nitrocellulose paper and detection by anti-BGH serum. This method was also served for semi-quantitative determination of BGH in the bacterial extract.

Animals↗

Expression of a cloned bovine growth hormone gene in Escherichia coli minicells.

The synthesis of polypeptides in Escherichia coli minicells, directed by a pBR322 plasmid and its derivative-carrying bovine growth hormone cDNA insert, was studied. Two polypeptides coded by the ampicillin-resistance (Apr) gene (32 000 and 28 000 daltons) and a tetracycline-resistance (Tcr) polypeptide (36 000 daltons) were identified by insertion inactivation. Two additional polypeptides of 37 000 and 34 000 daltons of as yet unknown function were detected in all extracts regardless of the presence of the Apr or Tcr genes in the plasmid. The pBR322-BGH recombinant plasmid coded for several novel polypeptides, among them one of 46 000 daltons, presumably a fused product of the BGH and beta-lactamase genes. This protein, however, was not secreted into the periplasmic space of the cells as was the beta-lactamase.

Animals↗

Cloning of bovine growth hormone gene and its expression in bacteria.

A hybrid plasmid was constructed containing beta-lactamase gene of plasmid pBR322 and cloned coding sequences of bovine growth hormone (BGH). The constructed plasmid contains all DNA sequences required to encode BGH, and when used as a hybridization probe it detects one growth hormone gene in the bovine genome. The cloned DNA sequences are inserted into the beta-lactamase gene in the correct reading frame for BGH synthesis. The hybrid gene is expressed in bacteria and the product, a fused beta-lactamase-bovine growth hormone protein, is specifically immunoprecipitated with anti-serum to BGH. Unlike beta-lactamase, very little growth hormone containing sequences can be detected in the periplasmic space.

Amino Acid Sequence↗

Adsorption of actinophage Pal 6 to developing mycelium of Streptomyces albus.

The adsorption of actinophage Pal 6 to mycelium of different ages was studied. The rate of phage adsorption to germinating spores increased with age, being maximal at about 4 h after start of germination. The number of plaque forming units (PFU) attached to mycelium had apparently decreased later, while the phage titer in the medium supernatant was also drastically reduced. These contradictory findings were explained by the increase in the adsorption capacity of the mature mycelium due to the formation of clumps, made of network structure which adsorbed many phages, but were counted as only 1 PFU each. The growth curve of the phage in fully developed mycelium and in germinating spores were compared.

Actinomyces↗

Isolation of viable deletion mutants of Streptomyces actinophage (Pal 6) and their molecular characterization.

Deletion mutants of bacteriophage Pal 6 were isolated by successive treatments of either heat (60 degrees C) or pyrophosphate (10 mM). These mutants were characterized by restriction enzyme cleavage analysis. The pyrophosphate resistant clones lost the whole Eco R1 fragment in which the Sal I site is located, as well as an unrelated Hind III fragment. These results show that the region containing the Sal I site in the phage genome is not essential for phage viability. This single Sal I site is therefore suitable as a potential insertion site for DNA cloning. On the other hand, the heat resistant clones that were isolated and characterized do not appear to have detectable deletions as indicated by their Eco R1 DNA digestion pattern.

Bacteriophages↗

Radioimmunoassay of gentamicin in Micromonospora medium extracts.

Medium extracts of Micromonospora adversely affected a radioimmunoassay which was used for the measurement of gentamicin in the medium. An overestmation or an underestimation resulted, as judged by the addition of extracts to a gentamicin standard sample.

Chromatography↗

Gentamicin bioautography assay vs. the microbiological disk test.

The use of bioautography for quantitative measurement of gentamicin concentrations was compared with the disk test. Following chromatographic separation and bioautography, gentamicin produced inhibition zones, 2 approximately 7 times larger than the inhibition zones formed by the same amounts of gentamicin in the disk test. Bioautography, therefore, is a more sensitive assay method.

Bacillus subtilis↗

Evaluation of several enrichment procedures for the isolation of recombinant plasmid DNA.

A number of methods for the selective enrichment of recombinant plasmids were examined; these include alkaline phosphatase treatment of the restricted pBR322 vector, as well as a combination of this and S1 nuclease treatment of the ligated mixture of pBR322 and pCR1 plasmids or S. griseus DNA followed by D-cycloserine treatment to enrich for cells carrying recombinant molecules. The relative efficiencies of these methods were compared.

Alkaline Phosphatase↗

Discoordination of ribosomal RNA metabolism during metabolic shifts of Spirodela plants.

The effects of metabolic shifts on nucleic acid syntheses have been widely studied in prokaryotes, but not in plants because of a paucity of suitable systems. Spirodela (Duckweed) was thus used to ascertain the response of the nucleocytoplasmic (nc) and plastid ribosomal RNA metabolisms to partial and total carbon deprivation. The 0.56 X 10(6) Mr plastid rRNA is the one species of RNA most affected by metabolic shifts; unlike other species, its appearance is delayed by deprivation and it appears more rapidly than other species on transfer from dark to light. The data suggest a discoordination between the transcription and processing of plastid ribosomal precursors. Incorporation into all nc and plastid rRNAs was severely reduced and all rRNA precursors accumulated in green plants that were completely deprived of carbon by transferring to the dark, without sucrose. The amounts of nc and plastid precursors transcribed readjusted to the reduced amounts processed to mature RNA only after long periods in the dark with sucrose. This delay involved the formation of new colorless plants. Less plastid RNAs, compared to nc RNAs are found in the dark steady state.

Chloroplasts↗

Polyacrylamide gel as a medium for DNA dissociation and reassociation.

Several properties of thermal denaturation and renaturation of DNA in polyacrylamide gels were investigated: (1) Following electrophoresis the DNA band was scanned and shown to increase in absorbance with increasing temperature. The increase was proportioned to DNA concentration across the peak. (2) The dependence of the Tm on salt concentration over a hundred fold range was similar to that found for DNA in free solution. (3) Denaturation of several DNA samples ranging in G + C content from 26 to 71% was compared in gels and free solution. The relationship between Tm and % G + C was virtually identical for both sets of DNAs. (4) The kinetics of DNA renaturation in the gel was followed. Reassociation of bacteriophage T4 DNA was 2nd order and proceeded more rapidly in polyacrylamide gels than in free solution.

Acrylamides↗

Control of lysine biosynthesis in Bacillus subtilis: inhibition of diaminopimelate decarboxylase by lysine.

Diaminopimelate decarboxylase has been characterized in extracts of Bacillus subtilis and resolved from aspartokinases I and II. Under certain conditions, the enzyme is specifically inhibited by physiological concentrations of L-lysine, but less specificity and altered kinetics of inhibition are observed if lower ionic strengths are employed in the assay procedure. Diaminopimelate decarboxylase can be desensitized to lysine inhibition by either lowering the pH or diluting the enzyme in Tris buffer in the absence of pyridoxal phosphate. Evidence is presented to incidate that, under proper conditions, lysine inhibition involves an interaction of the amino acid with the enzyme rather than competition for available pyridoxal phosphate in the assay. Lysine, by affecting the level of meso-diaminopimelate, may thus regulate its biosynthesis through sequential feedback inhibition. Analysis of the diaminopimelate decarboxylase of 15 revertants of mutants that had originally lacked diaminopimelate decarboxylase activity indicates that as little as 5% of the specific activity of enzyme observed in the wild-type strain is sufficient to permit normal growth rates. In the growing cell, diaminopimelate decarboxylase may therefore exist largely in an inhibited state.

Aspartic Acid↗