Search PubMed⌕ Search

Biomedical subjects

R J Reid

Publications and source records attributed to R J Reid.

53 records · Page 3Linked to original sources
PubMed1994

Probing the structure of bacteriophage phi 29 prohead RNA with specific mutations.

Bacteriophage phi 29 of Bacillus subtilis packages its double-stranded DNA genome into a preformed prohead in an ATP-dependent reaction. A 174-residue phi 29-encoded RNA molecule (pRNA) is a structural component of the prohead and is essential for DNA packaging. The secondary and tertiary structures of the prohead binding site on pRNA have been probed using a series of specific mutant pRNAs and by measuring binding to RNA-free proheads and in vitro packaging of the DNA-gene product 3 (DNA.gp3) complex. A pseudoknot in pRNA inferred from phylogenetic studies was confirmed with specific mutations, and this pseudoknot was necessary for DNA.gp3 packaging activity. pRNA was truncated progressively from the 5' and 3' ends to isolate the prohead binding site, and three truncated pRNAs of 79, 71, and 62 residues retained prohead binding activity but could not reconstitute proheads for DNA.gp3 packaging. Mutation resulting in changes of the D hairpin loop and its connecting residues within the prohead binding site of pRNA and DNA packaging studies demonstrated that some alteration of secondary structure in this helix was permissible. The analyses provided further confirmation of a discrete prohead binding domain in pRNA and further delineated specific structural requirements for DNA.gp3 packaging activity which may not be required for prohead binding.

Bacillus Phages↗
PubMed1994

Identification of bacteriophage phi 29 prohead RNA domains necessary for in vitro DNA-gp3 packaging.

Functional domains of the bacteriophage phi 29 prohead RNA (pRNA) that are essential for in vitro packaging of DNA-gp3 into the prohead were mapped using pRNA mutants. Oligonucleotide-directed mutant pRNAs were produced that contained deletions and sequence alterations but were predicted to retain the overall secondary structure of wild-type pRNA. Mutant pRNAs were compared to wild-type pRNA for prohead binding in a competition assay and for DNA packaging in the defined in vitro system. The prohead binding site was previously localized to residues 22-84 on the 120-residue domain I of pRNA by ribonuclease footprinting (Reid, R. J. D., Bodley, J. W., and Anderson, D. (1994) J. Biol. Chem. 269, 5157-5162). Mutations of pRNA within the prohead binding site resulted in substantial loss of prohead binding capacity, while mutations outside of the footprint had moderate effects on prohead binding. DNA-gp3 packaging activity was correlated with pRNA binding activity for mutations within the footprint. This mutational analysis showed that both sequence and secondary structure of residues 40-91 of pRNA were crucial for prohead binding and that elements of the A helix formed from residues 1-28 and 117-92 were needed for DNA packaging functions other than prohead binding.

Bacillus Phages↗
PubMed1994

Characterization of the prohead-pRNA interaction of bacteriophage phi 29.

The small prohead RNA (pRNA) of the Bacillus subtilis bacteriophage phi 29 is essential for ATP-dependent packaging of viral DNA. The 174-, 124-, and 120-residue forms of pRNA produced in vitro using T7 RNA polymerase were equivalent in prohead binding and DNA packaging activity to pRNAs produced in phi 29-infected cells. pRNA binding to proheads, characterized by the use of Northern hybridization and filter binding assays, was specific, rapid, and irreversible in the presence of 10 mM Mg2+. Proheads produced in phage-infected cells carried 5.8 +/- 2.7 copies of pRNA, and proheads assembled in Escherichia coli in the absence of pRNA bound 6.0 +/- 3.5 copies of pRNA. Footprints of proheads on pRNA generated with the ribonucleases A, T1, and V1 showed that nucleotides 22-84, 5' to 3', were protected from ribonuclease attack. Enhanced cleavage at nucleotides 37-40 with ribonuclease V1 suggested a conformational change of pRNA upon prohead binding.

Bacillus Phages↗
PubMed1993

The uptake and metabolism of urea by Chara australis: IV. Symport with sodium--a slip model for the high and low affinity systems.

We have previously investigated the electrogenic influx of urea in Chara, and the urea- and sodium-dependent membrane current. We have shown that there is a sodium-stimulated component of urea influx and a urea-stimulated component of sodium influx, and that these are of the same size. We conclude that the electrogenic inward transport of urea, and of its analogues acetamide and acrylamide, is by sodium symport, with a stoichiometric ratio of 1:1. The kinetics of the fluxes and currents show two different KM values for sodium in different cells and two different kinds of kinetics for the effect of urea on membrane current, one of which fits the Michaelis-Menten equation, while the other shows a maximum and fits the difference of two Michaelis-Menten terms, suggesting a phenomenon like cis-inhibition. Similarities in kinetic characteristics between the inhibitory site and the electrically silent uptake site (System II) lead us to suggest that the same protein may be responsible for both the low-KM, electrogenic influx of urea (System I) and the high-KM, electrically silent influx by System II. We suggest a "slip" model for urea uptake in Chara.

Acetamides↗
PubMed1992

Regulation of Calcium Influx in Chara: Effects of K, pH, Metabolic Inhibition, and Calcium Channel Blockers.

Measurements were made of (45)Ca influx into isolated internodal cells of Chara corallina and also into internodal cells of intact plants. (45)Ca influx was closely related to growth. In rapidly expanding internodal cells, the influx was approximately 1.4 nmol m(-2) s(-1) compared to the influx in mature cells from slow-growing cultures of 0.2 nmol m(-2) s(-1). Isolated internodal cells had influxes in the range 0.2 to 0.7 nmol m(-2) s(-1), but this increased to approximately 2 nmol m(-2) s(-1) in high calcium solutions and to 4 nmol m(-2) s(-1) in high potassium solutions. No significant effects on calcium influx were observed for changes in external pH or for treatments that changed internal pH, except that NH(4) was slightly inhibitory. Severe metabolic inhibition by carbonylcyanide-m-chlorophenyl-hydrazone stimulated influx, whereas dicyclohexylcarbodiimide had no effect and darkness inhibited influx. La(3+) also inhibited influx, but the organic channel blockers nifedipine and bepridil stimulated influx. Verapamil had no effect. The results are generally consistent with voltage regulation of calcium channels as in animal cells.

Journal Article↗
PubMed1992

Tissue responses to Hydron, assessed by intraosseous implantation.

Teflon tubes containing freshly mixed, polymerized Hydron root canal filling material, fully set AH26 or Teflon were implanted into the mandible of guinea pigs and assessed histologically at 2 days, 1, 2, 4, 12 and 26 weeks. None of the materials tested elicited signs of overt or significant tissue damage, and polymerized Hydron was assessed to be as biocompatible as fully set AH26 and Teflon. Bone formed in very close apposition to the polymerized Hydron, whereas a soft tissue capsule separated the regenerated bone from implants of AH26 and Teflon.

Animals↗
PubMed1992

Tissue responses to Hydron, assessed by intramuscular implantation.

Freshly mixed polymerized Hydron root canal filling material, fully set AH26 and Teflon were implanted in the quadriceps muscle of guinea pigs and assessed histologically at 2 days, 1, 2, 3, 12 and 26 weeks after implantation. All materials were characterized by peri-implant fibrous connective tissue capsule formation. Von Kossa-positive calcific material was observed at the implant-tissue interface of Hydron implants. The amount of apparently calcified material increased with time. Inflammation was not a prominent tissue response for any of the test materials, nor was a foreign body giant cell response.

Animals↗
PubMed1992

A five-year study of Hydron root canal fillings.

A prospective clinical and radiographic study was conducted in order to compare Hydron and laterally condensed gutta-percha/AH-26 root canal fillings. Paralleling technique periapical radiographs were taken preoperatively, postoperatively and at recall appointments at post-treatment intervals of 6 months, 1 year, 2 years, 3 years and 5 years. Clinical examination at the recall appointments revealed no adverse signs or symptoms amongst all the patients who attended (mean attendance 44.5% at each interval). Radiographs were scored according to the periapical status of the treated root, and comparable bone healing rates were observed between the two root-filling materials. Among the patients attending recall appointments, there were no radiographic signs of failure of any of the 39 gutta-percha/AH-26 root canal fillings. However, three of the 35 canals filled with Hydron were classified as failures, and four required further assessment after the 5-year recall appointment. This study indicated that Hydron and gutta-percha/AH-26 root canal fillings were well accepted but, on the basis of radiographic assessment, success with gutta-percha/AH-26 was more predictable.

Bismuth↗
PubMed1985

Pitfalls in, and new methods for, the radioimmunoassay for digoxin in urine.

Examining techniques for radioimmunoassay of digoxin in urine, we found that the apparent concentration of digoxin is affected by endogenous substances, independent of the concentration of digoxin itself. We describe a modification of the Becton Dickinson solid-phase RIA that makes it more nearly accurate and more easily performed, and obviates the endogenous interference. Digoxin concentrations from 5.0 to at least 1000 micrograms/L can be measured.

Digoxin↗
PubMed1981

Chemical modification of aminoglycosides. 3. Synthesis of 2"-deoxykanamycins from neamine.

The preparation of 2"-deoxykanamycin B (12) and 2",3',4'-trideoxykanamycin B (14) from neamine (1) is described. Key intermediates in the synthesis of these 2"-deoxyaminoglycoside antibiotics are 3',4'-bis-O-(p-nitrobenzoyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (6) and 3',4'-dideoxy-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (9). The amino groups of these intermediates are blocked by the trifluoroacetyl group, a blocking group not widely used in aminoglycoside chemistry.

Bacteria↗
PubMed1980

Hydralazine-pyrimidine interactions may explain hydralazine-induced lupus erythematosus.

Hydralazine, the prototypic drug that induces systemic lupus erythematosus, reacts with thymidine and deoxycytidine. Analysis of a reaction mixture of therapeutic concentrations of hydralazine with labeled thymidine reveals at least four labeled products. At higher concentrations, hydralazine reacts with labeled deoxycytidine to form at least three labeled products. Formation of these products is markedly enhanced by exposure to ultraviolet light. The reaction of hydralazine with thymidine and deoxycytidine may be in part responsible for initiating drug-induced systemic lupus erythematosus.

DNA↗
PubMedDate not supplied

Chronic meningoencephalitis associated with Brucella sp. infection in live-stranded striped dolphins (Stenella coeruleoalba).

A chronic, non-suppurative meningoencephalitis was found in three young striped dolphins (Stenella coeruleoalba) during routine neuropathological examination of marine mammals live-stranded on the Scottish coast. In all three dolphins the lesions were associated with the isolation of a Brucella sp. from the brain and with the immunohistochemical detection of brucella antigen. Moreover, antibodies to Brucella spp. were detected in the two dolphins that were subjected to serological examination. Immunohistochemical and serological examinations for morbillivirus antigen and antibodies, respectively, were negative in all cases. Although brucella infection of marine mammals has been extensively documented in recent years, its association with lesions and disease is less well recognized. The present report provides the first description of an association between Brucella sp. infection and neuropathological changes in a cetacean species.

Animals↗