Biomedical subjects
Publications and source records attributed to R L Himsworth.
We report an 84-year-old woman with hypoglycaemia and hyperinsulinaemia caused by diffuse nesidioblastosis. This is the oldest case of nesidioblastosis so far recorded. The case illustrates some of the difficulties in diagnosing inappropriate hyperinsulinaemia and the benefits of surgery, even in old age. Hypoglycaemia should be considered in patients with variable mental function.
We have identified a polymorphic tetranucleotide consisting of (AAAT)n within the first intron of the parathyroid hormone (PTH) gene, and have used this to investigate the segregation of the PTH gene and idiopathic hypoparathyroidism in 7 affected and 21 unaffected members from three families. An association between the PTH locus and autosomal dominant idiopathic hypoparathyroidism in one family was excluded by observing recombination between the two loci. In the remaining two families with autosomal recessive idiopathic hypoparathyroidism, the PTH locus was not similarly excluded. We had previously demonstrated a donor splice site mutation of the PTH gene in one of these families, and PTH gene abnormalities were therefore sought in the second of these families. DNA sequence analysis of the three exons, together with 4 exon-intron boundaries and the promoter region of the PTH gene revealed no abnormalities, thereby indicating molecular pathology at another locus. Thus, our analysis of idiopathic hypoparathyroidism reveals genetic heterogeneity for this disorder. In addition, our identification of a microsatellite polymorphism of the PTH gene should help further segregation studies of this locus in families with parathyroid disorders.
The heterogeneity of circulating TSH-receptor antibodies (TRAb) has been demonstrated directly by affinity chromatography on the dye Remazol Yellow-GGL. The technique resolved these antibodies into discrete peaks within the general serum immunoglobulin background. Peaks were detected initially by a binding (radioreceptor) assay and then characterized by their ability to stimulate the uptake of iodine-125 into FRTL-5 cells. Thirteen subjects with high serum levels of TRAb were studied. The two hypothyroid patients each produced a single peak of TRAb activity with no ability to stimulate uptake of iodine-125 into cultured cells. The remaining 11 patients had Graves' disease; their sera produced a range of elution profiles after chromatography with TRAb activity resolved into as many as five peaks. Considerable differences were seen in the relative abilities of the resolved TRAb components to bind to receptors and to stimulate cellular uptake of iodine-125 both between and within individual samples. No two patients produced components which chromatographed in the same position on the gradients employed for elution from the dye.
The characteristics of thyroglobulin (Tg) autoantibodies in Obese strain (OS) chickens with thyroiditis have been defined and compared with those of polyclonal antibodies to chicken Tg produced by immunizing normal chickens and a rabbit, and with mouse monoclonal antibodies (MoAb) to chicken Tg. Chicken Tg autoantibodies (aAb), when tested against Tg from 24 species all showed specificity for chicken Tg which ranged from absolute to limited although in most instances cross-reactions with Tgs of other species were either absent or at a low level. Antibodies to chicken Tg produced by immunization showed a similar limited range of cross-reactions. Four of five chTg-MoAbs were specific for chicken Tg and the fifth was almost so. In competitive experiments, the polyclonal rabbit antibody could fully inhibit binding of all chicken Tg-aAb to chicken Tg but not vice versa. It was inferred that polyclonal rabbit Tg antiserum includes antibodies to all the epitopes seen by chicken Tg-aAb and many more besides. In similar experiments with four chicken Tg-MoAbs, the binding of one chicken Tg-aAb was unaffected, and three other patterns of inhibitions were defined. The binding to chicken Tg of a fifth chicken Tg-MoAb was enhanced rather than inhibited by chicken Tg-aAb. Some but not all chicken Tg-aAb preparations could differentiate between Tgs containing different amounts of thyroxine. We conclude that the autoantibodies to Tg in OS chickens are directed in the main against determinants unique to the species. Not all the species-specific determinants are involved in the autoimmune response but the number of epitopes involved is at least four. In these respects the immune response to Tg in OS chickens resembles that in autoimmune thyroid disease in humans. The conformation of chicken Tg may be affected by combination with antibody or by the content of thyroid hormone.
Two female patients with functioning adrenocortical carcinomas had plasma ACTH detectable by RIA at presentation. In both patients there was evidence for biological activity of ACTH. There was no evidence for an ectopic source of ACTH, nor for a pituitary tumour. Urinary steroid analysis showed patterns of multiple hormone secretion characteristic of adrenocortical carcinomas. The finding of detectable ACTH concentrations in a patient with Cushing's syndrome does not exclude the presence of an adrenocortical tumour.
The T4-binding proteins of a euthyroid subject with persistent hyperthyroxinemia (T4, greater than 20 micrograms/dl) were present in normal concentrations. Abnormal transport of both T4 and rT3 was demonstrated by reverse flow paper electrophoresis; excess T4 was bound to albumin and prealbumin, while increased binding of rT3 was confined to prealbumin. The three T4-binding proteins in the serum of the subject were isolated by affinity chromatography and characterized. Equilibrium dialysis experiments demonstrated a 20-fold increase in affinity of the albumin for T4 (Ka, 5.1 X 10(6) M-1) and a 4-fold increase in affinity of prealbumin for T4 (Ka, 3.0 X 10(8) M-1); T4-binding globulin affinity was normal. Nine other members of the family were also studied. Two sisters of the propositus have both the abnormal albumin and the variant prealbumin, while a brother has normal T4-binding proteins. The mother has the abnormal albumin alone. The father, his sister, and one of his three brothers have the variant prealbumin only. Despite the presence of the variant prealbumin in some of the paternal relatives of the propositus, their total iodothyronine concentrations were within the normal ranges; the condition may, therefore, often go undetected. The characteristics of the albumin found in the affected members of this kindred are those we have defined for familial dysalbuminemic hyperthyroxinemia type I, which is inherited as an autosomal dominant trait. The pattern of inheritance of the variant prealbumin is also consistent with a dominant mode with strong penetrance. The presence of two separately inherited abnormal T4 transport proteins in the same family suggests that both conditions may be more common than has been thought.
Significant binding of thyroglobulin (Tg) autoantibodies was restricted to primate Tg, although some sera showed weak cross-reactivities with other species at high concentrations. In contrast rabbit anti-human Tg bound to all other animals' Tg in addition to human and this was not due to crossreactions with the thyroxyl residues. Mouse antihuman Tg monoclonal antibodies (MoAb) showed at least three different patterns of cross-reactivities. Group 1 MoAb, like Tg autoantibodies, were primate specific; two out of six MoAb bound only human Tg. Group 2 MoAb also bound a few other animals' Tg in addition to human. Of the three MoAb in group 3, two reacted with most or all of eight animal Tg tested while the other bound to all eight. However the binding of thirteen out of the fourteen MoAb against human Tg including all of the primate Tg specific group 1 MoAb was not inhibited by any of the nine Tg autoantibodies. Reactions of MoAb with heat-treated human Tg varied. Examples of decreased, increased or no change in reactivity could be demonstrated. On this basis at least five or possibly six separate human-specific epitopes could be defined by group 1 MoAb. Monoclonal antibody 1D6 (group 2) was inhibited by six out of nine Tg autoantibodies at high concentrations. This MoAb, similarly to the eight Tg autosera tested, had reduced reactivity with heat treated Tg. These studies demonstrated that human Tg has immunogenic structures conserved in the Tg of many species. The protein also has, however, primate and human-specific antigenic sites but at least five or possibly six of these are not related to autoantibody binding.
Explore the source record for details and available documents.
Thyroxine remains attached to its synthetic site in thyroglobulin until it is released by proteolysis. Strong homology in the primary sequence surrounding thyroxine-forming residues in thyroglobulins from various species suggests a unique three-dimensional structure at hormonogenic sites. To examine this, two thyroxine-binding mouse anti-(chicken thyroglobulin) monoclonal antibodies, 1A10 and 5F6, were used as probes for this region in an enzyme-linked immunosorbent inhibition assay. The thyroxine content of thyroglobulins had a marked positive influence on the monoclonal antibody binding: when the thyroxine content of human thyroglobulin rose by 6.6-fold, cross-reactivities rose 25-fold for the 1A10 monoclonal antibody and 17.6-fold for the 5F6 monoclonal antibody. However, interspecies comparison of thyroglobulin preparations with similar thyroxine content showed lower than expected cross-reactivities for human, pig and sheep thyroglobulins when compared with chicken thyroglobulin. Only when the thyroxine content of heterologous thyroglobulin preparations was two or three times higher did the cross-reactivities equal or surpass that of chicken thyroglobulin. It is concluded that in thyroglobulin there are structural differences in the different animal species near the thyroxine-forming sites bound by these monoclonal antibodies. The known primary sequence similarity does not seem to result, therefore, in identical three-dimensional structures about this site. These differences may reflect species-specific variations in distant regions brought close as a result of chain folding to form the hormonogenic site, such as those around the donor diiodotyrosine residue or in polysaccharide structures. These monoclonal antibodies provide information about the structure of thyroglobulin, which cannot be obtained from knowledge of the amino acid sequence alone.
Nonadherent, low density cells of dendritic morphology from the blood of patients with Hashimoto's thyroiditis were treated with human thyroglobulin (Tg) in vitro and cultured under serum-free conditions with autologous patient B cells and irradiated T cells. The patients were selected for high serum levels of IgG antithyroglobulin antibody (anti-Tg IgG). In 2 out of 12 patients the Tg-treatment induced production of anti-Tg IgG in excess of that secreted spontaneously. The amount of antibody produced in vitro (whether increased by Tg or not) correlated with the serum levels of antibody. In 5 patients (including the 2 who responded to Tg) the ratio of supernatant IgG anti-Tg antibody to total IgG was reduced when polyclonal stimulation was done with BCGF (10%). Antibody production was absent in cultures of cells from 2 patients with Graves disease and 4 normal individuals. Thus, in some patients with Hashimoto's thyroiditis, an extrinsically added autoantigen (Tg) on blood-derived dendritic cells can induce IgG anti-Tg antibody in vitro. These data suggest that "professional" antigen-presenting cells may play a role in autoimmune thyroid disease.
Serum concentrations of total and free T4 (TT4 and FT4), total and free T3 (TT3 and FT3), rT3, T4 binding globulin (TBG), T3 uptake (T3U) and TSH were measured in 12 patients with severe hypothyroidism before and during the introduction of replacement therapy with oral T4. The dose of T4 was increased by increments of 50 micrograms at intervals of 4 weeks to a total of 200 micrograms daily. There was a linear correlation between the concentrations of FT3 and FT4 (FT3 = 1.35 + 0.23FT4, r = 0.916, P less than 0.001). The correlation between TT3 and TT4 was more complex: the data were best fitted by the expression TT3 = 0.195 square root TT4, (r = 0.936, P less than 0.001). The relatively greater rise in TT3 initially may reflect a greater binding of T3 by TBG when the concentration of T4 is low. TBG concentration fell after 50 and 100 micrograms of T4 but did not change at the higher doses. There was a simple linear relation between TT4 and rT3 (rT3 = -0.022 + 0.0027TT4, r = 0.921, P less than 0.001). The expected inverse relation between TSH concentration and the thyroid hormones was seen, the three closest correlations being between the logarithm of the TSH concentration and FT3, the ratio T4/TBG and FT4 (r = 0.927, -0.917 and -0.900 respectively). These correlations were significantly better (P less than 0.05) than the correlations with untransformed TSH values. Suppression of TSH occurred while FT3 tended to remain within normal limits, but FT4 was often raised.(ABSTRACT TRUNCATED AT 250 WORDS)
Amiodarone was given by daily intraperitoneal injection (10 mg/kg body weight) to a group of 11 New Zealand white rabbits. Over 6 weeks there were significant increases (P less than 0.01) in plasma total T4 concentration (43 +/- 8 to 60 +/- 13 nmol/l; mean +/- 1 SD) and rT3 concentration (0.12 +/- 0.06 to 0.31 +/- 0.16 nmol/l) and a significant fall in total plasma T3 (2.3 +/- 0.3 to 1.7 +/- 0.2 nmol/l). The plasma clearance of T4 in rabbits treated with amiodarone for 6 weeks was significantly reduced relative to controls (64 +/- 25 vs 109 +/- 19 ml/kg per day, P less than 0.01) but the production rate was unchanged (3.8 +/- 1.8 vs 3.7 +/- 1.4 nmol/kg per day). In the amiodarone treated animals the increased plasma T4 concentration was entirely accounted for by the reduced clearance. It is inferred that the normal output of T4 by the thyroid is pituitary dependent and that TSH secretion is sustained in the presence of hyperthyroxinaemia because of amiodarone induced partial inhibition of T4 utilization by the thyrotroph.
A series of human autoantibodies against thyroglobulin (Tg) which exhibit different specificities for iodothyronines were studied. The ability of a thyroxine (T4)-containing peptide (T4P) isolated from human thyroglobulin (Tg) to displace [125I]T3 from human T3-specific autoantisera was 11-50 times greater than that of T4 alone. These antisera therefore strongly recognize amino acids adjacent to T4 in the Tg structure. This was confirmed when a Tg preparation (Tg[0.05]) containing an average of only 0.05 of a T4 residue/molecule and much less T3 had good cross-reactivities with these antisera. Cross-reactivities of other Tg preparations with different T4 contents increased only slowly with increase of T4 content up to a mean of 6.6 residues/molecule and were not proportional to T3 content. In contrast, cross-reactivities with a human T4-specific autoantiserum were strongly dependent on T4 content. Tg[0.05] was 500 times less reactive than T4P and 615 times less than T4. Cross-reactivities rose rapidly as the T4 content of Tg preparations increased from a mean of 0.05 to approx. 1-2 residues/molecule. Thyroxine is therefore a dominant feature of the antigenic site for this antiserum. There was little further increase in cross-reactivities for those Tg preparations containing up to an average of 6.6 residues T4 per molecule, confirming previous conclusions that all T4-containing sites are not immunologically identical and that autoantibodies exhibit a preference for particular sites on Tg. Similar conclusions were reached for a non-specific iodothyronine-binding antiserum. These results indicate that iodothyronine specificity in human autoantisera is not necessarily determined by the iodothyronine present in the immunogenic area, but by the precise site selected by the immune response. T4- or non-specific antibodies have thyroxine as a dominant feature of the antigenic site. T3- specific antibodies have the thyroxine residue as a peripheral feature of the binding site, and it is not necessary to postulate that T3 was part of the immunogen or is required in the epitope. These antisera may have value in mapping the hormonogenic regions in Tg from human and other species.
Explore the source record for details and available documents.
A new variant albumin with increased affinities for iodothyronines has been identified. A euthyroid woman had raised total serum concentrations of T4 (155 nmol/l), T3 (3.0 nmol/l) and rT3 (700 pmol/l) but normal levels of all three iodothyronine-binding proteins. The affinity constant for T3 binding to the albumin was substantially raised (2.2 x 10(5) l/mole; normal immeasurable), that for rT3 (1.4 x 10(6) l/mole) was increased three-fold. This new albumin binds the analogues of T4 and T3 used in Amerlex free-hormone assays more strongly than does normal albumin, resulting in erroneously elevated estimates of serum free-T4 and free-T3 by this method. The new variant albumin was indistinguishable from normal albumin in molecular size and by electrophoretic and immunological techniques. Three distinct variant albumins exhibiting differential binding of iodothyronines have now been defined: Type I causes a raised total serum T4 only; Type II produces increased total T4 and rT3; Type III (the present example) results in elevated total T4, rT3 and T3. All three variants have normal free-T4 by dialysis but spuriously raised results by the Amerlex free-T4 method. Type III also causes an artificial increase in Amerlex free-T3. The pattern of thyroid function test results in Type III can readily be confused with both hyperthyroidism and with partial peripheral resistance to thyroid hormones.
A retrospective analysis of 594 unselected cases of hyperthyroidism from one district was made to determine whether the thyroid gland had been palpable on presentation. The thyroid had been diffusely palpable in 374 patients (63%), nodular in 110 (19%) and impalpable in 96 (16%); there was no clinical record in 14 (2%) cases. Under 40 years of age a diffuse gland was usual, being present in 86% of cases. The proportion of nodular and impalpable glands both increased progressively with age comprising 30% and 48% respectively in patients older than 70 years. Grave's ophthalmopathy was present in 13% of hyperthyroid patients with diffuse glands, in 12% of those with impalpable glands, but in only 1.5% of those with nodular thyroids. Thyroid autoantibodies were found with a similar frequency in all three groups. There was a very good correspondence between clinical assessment of shape and nodularity of thyroid and the results of isotope scanning; the majority of patients with impalpable glands (86%) had a diffuse uptake of isotope. In the elderly hyperthyroid patient an impalpable thyroid gland is common and is not a factor to weight against the diagnosis.
Turnover studies of thyroxine (T4), 3,5,3'-tri-iodothyronine (T3) and 3,3',5'-tri-iodothyronine (rT3) have been performed in the rabbit. A novel modification of a conventional radioimmunoassay has been used to measure specific 125I-labelled iodothyronines in small volumes of plasma in the presence of other 125I-labelled metabolites. Kinetic analysis of plasma disappearance of tracer was performed by a new theoretical approach. For T4 the mean (+/- S.D.) plasma concentration, clearance and production rates were 34 +/- 12 nmol/l, 109 +/- 19 ml/kg per day and 3.7 +/- 1.4 nmol/kg per day respectively (n = 9). For T3 the corresponding values were 2.04 +/- 0.42 nmol/l, 1.52 +/- 0.29 litres/kg per day and 3.07 +/- 0.76 nmol/kg per day (n = 8), and for rT3 0.12 +/- 0.04 nmol/l, 5.7 +/- 1.7 litres/kg per day and 0.69 +/- 0.23 nmol/kg per day (n = 8). The combination of these two new methodologies affords a simple and convenient means of studying iodothyronine metabolism under normal and abnormal conditions. The techniques employed may be generally applied to turnover studies of other compounds of physiological interest which can be measured by radioimmunoassay.
Explore the source record for details and available documents.