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Biomedical subjects

J R Walker

Publications and source records attributed to J R Walker.

At least 163 records · Page 9Linked to original sources

Bacteriophage lambda mutants (lambdatp) that overproduce repressor.

Lambda tp mutants, selected for their ability to form turbid plaques on lon hosts, overproduce repressor. The tp1 and tp2 mutations have been located within (or adjacent to) the cIII gene. The tp1 mutation reduced late gene expression, as measured by endolysin synthesis (in the absence of functional cI repressor) and progeny phage yield. The tp4 mutation was mapped in the cY-cII region, and complementation tests indicated that tp4 affects the diffusible product of the cII gene. The tp4 mutation also reduced progeny production, but did not markedly affect endolysin synthesis.

Coliphages↗

Bacteriophages in live virus vaccines: lack of evidence for effects on the genome of rhesus monkeys.

Four juvenile rhesus monkeys were inoculated with 10(12) plaque-forming units of the bacteriophage phiV1 isolated from live virus vaccines. After phiV1 had been cleared from the blood, DNA's were isolated from the livers and kidneys and analyzed for the presence of bacteriophage by plaque assays, and for the presence of phiV1 DNA by DNA-DNA reassociation kinetics. No evidence was found for persistence of the bacteriophage or for replication of the phage genome in these rhesus monkeys.

Animals↗

Prostaglandins and leucocyte migration in inflammatory reactions.

The effects of a variety of particulate irritants on the prostaglandin content, protein concentration and the leucocyte counts in the exudate of implanted inert sponges and on the development of paw oedema in the rat have been studied. Variations in prostaglandin accumulation and leucocyte migration were not related but irritants causing a long-standing paw oedema increased both parameters.

Animals↗

Isolation and characterization of plaque-forming lambdadnaZ+ transducing bacteriophages.

The Escherichia coli dnaZ gene, a deoxyribonucleic acid (DNA) polymerization gene, is located 1.2 min counterclockwise from purE, at approximately min 10.5 on the E. coli map. From a lysogen with lamdacI857 integrated at a secondary attachment site near purE, transducing phages (lambdadnaS+) that transduced a dnaZts (lambda+) recipient to temperature insensitivity (TS+) were discovered. Three different plaque-forming transducing phages were isolated from seven primary heterogenotes. Genetic tests and heteroduplex mapping were used to determine the length and position of E. coli DNA within the lambda DNA. Complementation tests demonstrated that the deletions in all three strains removed both att P and the int gene, i,e., DNA from both prophage ends. Heteroduplex mapping confirmed this result by demonstrating that all three strains had deletions of lambda DNA that covered the b2 to red region, thereby removing both prophage ends. Specifically, the deletions removed lambda DNA between the points 39.3 to 66.5% of lambda length (measured in percent length from the left and of lambda phage DNA) in all three strains. The three strains are distinct, however, because they had differing lengths of host DNA insertions. These phages must have been formed by an anomalous procedure, because standard lambda transducing phages are deleted for one prophage end only. In lambdagal and lambdabio strains, the deletions of lambda DNA begin at the union of prophage ends (i.e., position 57.3% of lambda length) and extend leftward or rightward, respectively (Davidson and Szybalski, in A, D. Hershey [ed.], The Bacteriophage Lambda, p. 45-82, 1971). Models for formation of the lambdadnaZ+ phages are discussed.

Chromosome Mapping↗

Physiological effects of growth of an Escherichia coli temperature-sensitive dnaZ mutant at nonpermissive temperatures.

The physiological effects of incubation at nonpermissive temperatures of Escherichia coli mutants that carry a temperature-sensitive dnaZ allele [dnaZ(Ts)2016] were examined. The temperature at which the dnaZ(Ts) protein becomes inactivated in vivo was investigated by measurements of deoxyribonucleic acid (DNA) synthesis at temperatures intermediate between permissive and nonpermissive. DNA synthesis inhibition was reversible by reducing the temperature of cultures from 42 to 30 degrees C; DNA synthesis resumed immediately after temperature reduction and occurred even in the presence of chloramphenicol. Inasmuch as DNA synthesis could be resumed in the absence of protein synthesis, we concluded that the protein product of the dnaZ allele (Ts)2016 is renaturable. Cell division, also inhibited by 42 degrees C incubation, resumed after temperature reduction, but the length of time required for resumption depended on the duration of the period at 42 degrees C. Replicative synthesis of cellular DNA, examined in vitro in toluene-permeabilized cells, was temperature sensitive. Excision repair of ultraviolet light-induced DNA lesions was partially inhibited in dnaZ(Ts) cells at 42 degrees C. The dnaZ(+) product participated in the synthesis of both Okazaki piece (8-12S) and high-molecular-weight DNA. During incubation of dnaZ(Ts)(lambda) lysogens at 42 degrees C, prophage induction occurred, and progeny phage were produced during subsequent incubation at 30 degrees C. The temperature sensitivity of both DNA synthesis and cell division in the dnaZ(Ts)2016 mutant was suppressed by high concentrations of sucrose, lactose, or NaCl. Incubation at 42 degrees C was neither mutagenic nor antimutagenic for the dnaZ(Ts) mutant.

Alleles↗

Inhibition of bacteriophage M13 and phix174 duplex DNA replication and single-strand synthesis in temperature-sensitive dnaZ mutants of Escherichia coli.

A functional dnaZ product, known to be essential for host DNA polymerization and for the synthesis of M13 and phiX174 parental replicative-form (RF) DNA, is required also for RF replication and single-strand synthesis by both of these phages. All three stages of M13 and phiX174DNA replication (parental RF formation, RF replication, and single-strand synthesis) are inhibited in dnazts mutants at elevated temperatures. In addition, the thermolabile step in M13 parental RF formation appears to occur after RNA priming;i.e., the synthesis of M13 RF DNA proceeded when a dnaZts mutant, infected at a nonpermissive temperature, was transferred to a permissive temperature in the presence of rifampin.

Coliphages↗

Anti-inflammatory drugs, prostaglandins and leucocyte migration.

The action of some non-steroidal acidic anti-inflammatory drugs, aspirin, phenylbutazone and indomethacin, in reducing leucocyte migration into the exudates of inert porous sponges implanted subdermally in the rat has been shown to be distinct from their effect in reducing the content of prostaglandins in the exudates. It is concluded that a component of the anti-inflammatory and antirheumatic actions of the drugs is concerned with a mechanism other than inhibition of prostaglandin biosynthesis.

5,8,11,14-Eicosatetraynoic Acid↗

Platelets, prostaglandins and inflammation.

In exudates of implanted sponges in rats, made thrombocytopenic by the administration of anti-platelet serum, there are significant reductions in the platelet and leucocyte counts and of the content of prostaglandin-like activity. It is concluded that platelets migrate into the developing sponge exudates, are the source of the prostaglandins and interact with complement to cause leucocyte migration. In normal animals the administration of indomethacin and sodium salicylate cause similar effects to thrombocytopenia whereas the injection of human plasma fraction affects only the leucocyte migration. One of the sites of the anti-inflammatory action of conventional non-steroidal drugs may be concerned with the migration of platelets into inflammatory lesions.

Animals↗

Prostaglandins and leucotaxis.

Freshly prepared solutions of prostaglandin E1 exhibit chemotactic activity against rabbit polymorphonuclear leucocytes harvested from the peritoneal cavity but are devoid of such activity either against similar cells obtained from the rat or against polymorphonuclear leucocytes obtained from the blood of the rabbit, rat and man. Some implications of these findings with respect to the development of inflammatory responses and the mode of action of non-steroidal acidic anti-inflammatory drugs are discussed.

Animals↗