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Biomedical subjects

C H McIntosh

Publications and source records attributed to C H McIntosh.

81 records · Page 5Linked to original sources
PubMed1977

Effect of dialysate calcium concentration on plasma parathyroid hormone during hemodialysis.

The effect of changes in the dialysate calcium concentration on calcium fractions and parathyroid hormone species in plasma during hemodialysis has been examined. C-terminal immunoreactive parathyroid hormone was suppressed when plasma calcium increased by greater than 25% whereas N-terminal fragments demonstrated an increase. The significance of this is discussed in light of present knowledge of parathyroid hormone metabolism.

Adult↗
PubMed1976

Radioimmunoassay of thyroxine-binding globulin in human plasma.

A highly specific and precise radioimmunoassay for thyroxine-binding globulin (TBG) has been developed. Crossreactivity with albumin and prealbumin was excluded. The normal range in young adults was 0.97 mg/100 ml. In childhood TBG was elevated (1.34 mg/100 ml) also in old age (1.28 mg/100 ml). In normals and in childhood there was a good correlation of TBG with thyroxine (T4). T4 did not correlate with TBG with regard to age. Triiodothyroxine (T3) did not correlate with TBG in any group. T4 and T3 exhibited a progressive decrease with age. No correlation between age and TBG was found. In mild thyrotoxicosis (T3 : 4.5 ng/ml, with normal T4 and negative TRH-test) TBG was slightly increased (1.20 mg/100 ml), whereas in more severe hyperthyroidism (T3 : 6.4 ng/ml, T4 : 16.3 mug/100 ml) TBG concentration was not significantly different from normals. In hypothyroidism TBG was elevated (1.26 mg/100 ml). The conclusion from these data is that TBG does not follow the progressive decrease of T4 and T3 with age. Thus, age-dependent euthyroid ranges for thyroid hormone concentration, including TBG concentration, must be established for better clinical discrimination of thyroid function. Possible dependence of TBG on the nature of thyroid hormone concentration must be considered in the production and peripheral kinetics of thyroid hormones.

Cross Reactions↗
PubMed1976

Characterization of the parathyrin receptor in renal plasma membranes by labelled hormone and labelled antibody binding techniques.

The parathyrin receptor in renal cortex has been investigated by studying the binding of 125I-labelled parathyrin, or of unlabelled parathyrin detected with 125I-labelled antibodies, to a partially purified plasma membrane fraction. The kinetics of hormone uptake demonstrated a biphasic response in both systems at 22 degrees C but this phenomenon was not detectable at 37 degrees C. Specific displacement of lactoperoxidase labelled 125I-labelled parathyrin occurred with 8 ng unlabelled bovine parathyrin. The apparent affinity constant was 2.3-10(8) M(-1) and the apparent binding capacity of the membranes 1.25 pmol/mg protein. Using the labelled antibody technique the receptor showed maximal binding at pH 7.0-7.5. As little as 80 pg bovine parathyrin produced a significant increase in binding of labelled anti-bovine parathyrin antibody and saturation of binding sites was demonstrated at 2.5 pmol/mg protein. Oxidized hormone showed undetectable binding. Treatment of membranes with phospholipases A or D, or Trypsin greatly reduced subsequent hormone binding. Prior incubation of membranes with 1-34 synthetic parathyrin decreased the binding of intact hormone whereas gastrin, insulin and glucagon had no effect. Growth hormone and calcitonin slightly increased parathyrin binding.

Animals↗
PubMed1975

New aspects of radioimmunochemical measurement of human parathyroid hormone using the labelled antibody technique.

Two forms of heterogeneity of parathyroid hormone (PTH) have given rise to conflicting results: one due to the heterogeneity of the secreted species from the gland and their peripheral metabolism and the other representing the immunochemical heterogeneity of the available antibodies. We have developed sequence specific assays using the technique of labelled antibodies. Therefore, results of assays measuring the C-terminal part and the (1-34)-N-terminal part of the molecule could be compared to those of an assay for hormone bearing both N- and C-terminal antigenic determinants. This assay is supposed to detect predominantly (1-84)-intact hormone. The immunoradiometric assay of (1-34)-PTH has a sensitivity of 0.04 ng/ml. This technique avoids the critical iodination of the hormone fragment containing no tyrosine. There is the expected overlap between normal subjects and patients with primary and secondary hyperparathyroidism. The most important finding are results from patients undergoing neck catheterization. We demonstrated nonuniform secretion of different species of PTH by parathyroid adenomata and normal glands. This supports the hypothesis of cleavage of the (1-84)-molecule in the gland.

Antigen-Antibody Reactions↗
PubMed1973

Multiple forms of acetylcholinesterase from pig brain.

1. A number of methods of solubilization of pig brain acetylcholinesterase (EC 3.1.1.7) were studied. The multiple enzymic forms of the resultant preparations were examined by polyacrylamide-gel electrophoresis. 2. Butanol extraction, Nagarase treatment and ultrasonication proved unsuitable as preparatory methods, but detergent treatment (Triton X-100, Triton X-100-KCl and lysolecithin) gave good yields. 3. Separation of soluble enzyme in three systems of polyacrylamide-gel electrophoresis were compared and the relative advantages are discussed. 4. By using a 6% (w/v) gel and continuous buffer system two forms of acetylcholinesterase were detected in Triton X-100-solubilized enzyme, but the incorporation of a sample and spacer gel and a discontinuous buffer system resolved this into four components. The forms of the soluble enzyme extracted by different methods differed in mobility. 5. With gradient polyacrylamide-gel electrophoresis between two and six forms were detected, depending on the method used for extraction. The average molecular weights of the five forms most frequently found were 60000, 130000, 198000, 266000 and 350000. 6. Treatment of the Triton X-100-extracted enzyme with 2.5m-urea altered the pattern and evidence of dissociation was observed. 7. The results are discussed in the light of present theories on the molecular structure of acetylcholinesterase.

Acetylcholinesterase↗
PubMedDate not supplied

VIP-innervation of rat intestine: a developmental study with monoclonal antibodies.

Four monoclonal antibodies to VIP have been generated and shown to be N-terminal specific with high affinity for VIP. VIP-containing nerve fibers and cell bodies were visible in the upper small intestine from day 1 of neonatal life. Initially the immunoreactivity was mostly in the myenteric plexus but extended into the sub-mucous plexus by day 7. From day 1 to day 7 the VIP-innervation developed both orally and caudally at a similar rate. In the stomach, the antrum showed sub-mucosal cell bodies by day 14, while in the corpus the cell bodies remained confined to the myenteric plexus. The colon showed positive fibers in the myenteric plexus at day 7 and cell bodies and fibers in the sub-mucous plexus by day 14. The size (cross-sectional area) of the individual VIP-immunoreactive cell bodies increased significantly between day 1 and day 14 with no further increase with age. At no time were immunoreactive cell bodies shown to migrate from the myenteric to the sub-mucous plexus. VIP-immunoreactive epithelial cells were not detected in the present study.

Aging↗
PubMedDate not supplied

Expression of recombinant human glucose-dependent insulinotropic polypeptide in Escherichia coli by sequence-specific proteolysis of a protein A fusion protein.

Glucose-dependent insulinotropic polypeptide (GIP) is a forty-two amino acid hormone that stimulates the secretion of insulin from the pancreatic B-cells in the presence of elevated glucose concentrations. The human GIP gene with the human A alpha-fibrinopeptide sequence was synthesized and linked to the Staphylococcus aureus protein A gene in the vector pRIT2T. This plasmid was expressed in Escherichia coli, and the resulting fusion protein consisted of three domains: protein A for ease of purification, fibrinopeptide sequence for thrombin cleavage and human GIP. The GIP was subsequently cleaved from the fusion protein with alpha-thrombin. The identity of the recombinant human GIP was confirmed by SDS-PAGE, ELISA, HPLC and amino-terminal amino acid sequence analysis. This recombinant product was shown to have comparable insulinotropic activity to porcine GIP in the isolated perfused pancreas.

Amino Acid Sequence↗