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Biomedical subjects

P England

Publications and source records attributed to P England.

42 records · Page 3Linked to original sources
PubMed1973

Increased thyroxine secretion following administration of dinitrophenol to rats.

1. Feeding male rats 0.2% dinitrophenol (DNP) in the diet caused weight loss, a significant fall in the plasma protein bound iodine concentration, and a rise in the thyroxine secretion rate. Thyroxine concentration was raised in the liver and intestinal tract with contents, reduced in the carcass, and unchanged in the pelt.2. Radio-iodine uptake by the thyroid gland was unchanged but there was a significant fall in the thyroid/serum activity ratio.3. Five hours after tracer radiothyroxine, DNP treated rats had a reduced fraction of dose in the plasma, with raised concentrations in the liver and intestines.4. Fractional turnover rates of radiothyroxine in plasma, liver and carcass were raised in DNP treated animals.5. DNP caused a significant increase in the biliary clearance of radiothyroxine, when measured immediately after administration of tracer and 4 hr later.6. It is postulated that DNP displaces thyroxine from its plasma binding proteins and that the displaced hormone is excreted in the bile. This wastage together with increased demand resulting from DNP induced hypermetabolism results in increased output of thyroxine by the thyroid gland.

Animals↗
PubMedDate not supplied

Recognition of E. coli tryptophan synthase by single-chain Fv fragments: comparison of PCR-cloning variants with the parental antibodies.

The use of a recombinant antibody fragment instead of a complete antibody, as a conformational probe for protein structure and folding studies, can be technically advantageous provided that the recombinant fragment and its parental antibody recognize the antigen through the same mechanism. Monoclonal antibodies mAb19 and mAb93 are directed against the TrpB2 subunit of Escherichia coli tryptophan synthase and they have been extensively used as conformational probes of this protein. DNA sequences coding for single-chain variable fragments (scFv) of mAb19 and mAb93 were cloned and assembled by reverse transcription of the mRNAs from hybridomas and PCR amplification. A specialized plasmid vector, pFBX, was constructed; it enabled to express the scFvs as hybrids with the maltose-binding protein (MalE) in E. coli, and to purify them by affinity chromatography on cross-linked amylose. Six independent clones were sequenced for each hybridoma. All of them had differences in their nucleotide and amino acid sequences. A competition ELISA and the BIAcore biosensor apparatus were used to compare the energetics and kinetics with which the parental antibodies and the hybrids bound TrpB2. The antigen binding properties of the hybrids were close to those of the parental antibodies and they were only weakly affected by the differences of sequence between the clones, with one exception. The stability of one of the hybrids and its antigen binding properties were strongly modified by a change of Gln6 into Glu, introduced into its VH domain by the PCR primers. Simple models of bimolecular interaction did not fully account for the kinetic profiles obtained with the parental antibodies and the hybrids, and this complexity suggested the existence of a conformational heterogeneity in these molecules.

ATP-Binding Cassette Transporters↗
PubMedDate not supplied

Paternity exclusion in koalas using hypervariable microsatellites.

Koala microsatellite loci containing the dinucleotide motif (CA)n were isolated from a size-fractionated (250-500 bp) koala genomic library and sequenced. Six locus-specific primer pairs were designed and synthesized for DNA amplification using the polymerase chain reaction (PCR). Microsatellite genotyping of 12 individuals generated unique "fingerprints" for each koala. All six microsatellite loci were polymorphic, with a mean of 6.5 +/- 0.6 alleles/locus. This level of allelic diversity is capable of generating > 4 x 10(9) DNA profiles, making it the most powerful technology for fingerprinting koalas currently available. Observed heterozygosities (H(o)) in the eight unrelated individuals surveyed ranged from 0.25 to 0.75, with a mean of 0.54 +/- 0.06. Mendelian inheritance of the observed polymorphism was confirmed by family studies. We demonstrate that microsatellite loci are ideal genetic markers for paternity exclusion and pedigree analysis of koalas, which have shown little genetic variation using most other methods.

Animals↗