Biomedical subjects
Publications and source records attributed to R J Thompson.
An antiserum specific for the brain-type aldolase C subunit has been used to investigate the cellular localization of this protein in human brain. Immunoperoxidase labeling at the light microscope level showed heavy staining of Purkinje cells in the cerebellum and of astrocytes in the cerebrum. Faint staining of occasional large neurons in the gray matter of the cerebral cortex was also seen.
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Clinicians have suggested that in some cases normal children have been inappropriately labeled as deviant by their parents and taken to psychological clinics for treatment. Reasons given for such inappropriate labeling have included factors such as the parents' marital distress and intolerance of normal child behavior. This study provided an empirical examination of the appropriateness or inappropriateness of parent labeling of 5--12-year-old children referred by their parents for treatment of conduct problems. The definition of inappropriate labeling derived by the present investigators required that the relationship between the child's actual behavior and the parent's negative label be so imperfect as to raise questions about the justification for the label, and that some identifiable factors other than the child's behavior account for the negative label. Three parent factors evaluated for their contribution to the parent negative label were martial distress, parent negative behavior toward the child, and parent distress about child deviant behavior. The statistical model of hierarchical multiple regression permitted analysis of this definition. The association found between observed child deviant behavior and the parent negative label suggested that parents of this sample perceived their children reasonably accurately. The three parent factors were found to be negligibly related to the parent label. Therefore, the conditions for inappropriate labeling of the children by the parents of this sample were not met.
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The background factors, problems, findings, and recommendations emerging from the interdisciplinary evaluation of 301 children referred to the Duke Developmental Evaluation Center for assessment of suspected development disabilities are analyzed. The perspective on developmental disabilities gained from utilization of a problem-oriented clinical record and a functional rather than a categorical approach is discussed. Particular attention is directed toward the frequency of findings in various dimensions of functioning and the implications arising from this comprehensive analysis for the roles of pediatricians, parents, and teachers in case management.
A protein has been purified from human brain that appears to be the human equivalent of bovine 14-3-3 protein. On polyacrylamide gel electrophoresis the protein migrates as a faster major component, termed 14-3-3-2 protein, and a slower minor component, termed 14-3-3-1 protein, which consists of approximately 12% of the total protein. Both 14-3-3-1 and 14-3-3-2 have a native molecular weight of approximately 67,000. 14-3-3-2 appears to have the subunit composition alpha beta; 14-3-3-1 has the composition beta'beta'. Peptide mapping with Staphylococcus aureus V8 proteinase shows that alpha and beta subunits are unrelated but the beta and beta' subunits show some common peptides. Immunoperoxidase labelling shows that 14-3-3 is localised in neurones in the human cerebral cortex. 14-3-3 shows no enolase, creatine kinase, triose phosphate isomerase, ATPase, cyclic nucleotide-dependent protein kinase, or purine nucleoside phosphorylase activity. 14-3-3 does not bind calcium and does not appear to be related to calmodulin, calcineurin, tubulin, neurofilament proteins, clathrin-associated proteins, or tropomyosin. The functional significance of this neuronal protein remains obscure.
An antiserum to human 14-3-3 protein has been produced in rabbits. The protein was a poor antigen and attempts to improve immunogenicity were unsuccessful. A radioimmunoassay was developed using the antiserum, 125I-14-3-3-2, and unlabelled 14-3-3-2 as standards. The assay had a sensitivity limit of 2.5 ng.ml-1. The minor component of human 14-3-3 protein (14-3-3-1 protein) cross-reacted to approximately 10% in the assay. Human tissues were surveyed for 14-3- protein by two-dimensional electrophoresis and by radioimmunoassay. Two-dimensional electrophoresis showed a 14-3-3 protein complex in brain, intestine, and testis, but not in other tissues. Radioimmunoassay showed that although brain had the highest concentration of 14-3-3 (13.3 microgram.mg-1 soluble protein), immunoreactivity was present in all tissues, with the concentration in intestine and testis approaching 50% of the brain level. Lower levels (less than 1.0 microgram.mg-1 soluble protein) were seen in liver, kidney, skeletal muscle, and erythrocytes. The immunoreactivity present in tissues other than brain showed the same molecular weight and charge characteristics and authentic 14-3-3 protein. The radioimmunoassay also detected 14-3-3 protein in serum (50 ng.ml-1) and in CSF (5-130 ng.ml). The immunoreactivity present in CSF appeared to be intact 14-3-3 protein. CSF 14-3-3 levels were measured in 82 patients with various neurological disorders. Measurements of this protein did not appear sufficiently discriminating to be of diagnostic value.
We used the BASODALT system (Cell 12: 1133--1142, 1977) of high-resolution two-dimensional gel electrophoresis to resolve proteins in extracts of human brain and, for comparison, those in nine other human organs. At least 180 protein-staining spots are visible on a BASODALT electrophoretogram of water-soluble human brain proteins. By electrophoresing extracts of different human organs, separately and mixed with brain extract, we could detect 14 basic spots that appear to be present in brain in concentrations at least 20-fold those in any other organ. Measurement of these new basic brain-predominant proteins may provide useful diagnostic information in neurological disorders.
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A two-site immunoradiometric assay for myocardial creatine kinase MB isoenzyme is described. The method utilizes immobilized anti-human creatine kinase BB antibodies and 125I-labelled anti-human creatine kinase MM antibodies and can specifically detect creatine kinase MB in the presence of approximately 1000-fold excess of creatine kinase MM or BB. Native creatine kinase MB prepared from human heart and creatine kinase MB prepared by hybridisation of purified human creatine kinase MM and creatine kinase BB appeared to react identically in the assay. Serum estimations on patients with suspected myocardial infarction correlated with the presence of an MB band on electrophoresis but preliminary results suggest that the two-site immunoradiometric assay may be more sensitive.
High-resolution multiple two-dimensional polyacrylamide gel electrophoresis (ISODALT) has been used to analyse soluble protein extracts from human brain and 12 other human organs. Approximately 200 protein gene products can be visualised on an electrophoretogram of soluble human brain proteins. By electrophoresing extracts of different human organs separately and mixed with brain extract, 8 proteins have been found which appear to be present in brain in concentrations at least 20 times greater than in any other organ. Four of these brain-specific proteins have been identified by co-electrophoresis with purified proteins as 14-3-2 protein, creatine kinase-BB isoenzyme, aldolase C4 isoenzyme, and 14-3-3 protein. The identities of the remaining 4 proteins are unknown.
Nine children who had enteroviral meningitis during the first three months of life and nine matched control children were evaluated for possible sequelae of their infection. The mean age of the children with meningitis was 46.67 months at the time of testing and that of the control subjects was 47.33 months. The mean IQ level of 89.67 obtained in the control children. Receptive vocabulary testing suggested that the receptive language functioning of the group with meningitis was significantly less than that of the control group. There was no significant difference in head circumference, no detectable sensorineural hearing loss, and no detected difference in intellectual functioning between the meningitis group and matched control subjects.
Indirect immunoperoxidase labelling at the light microscope level using a specific antisera to brain type creatine kinase-BB isoenzyme has localized the protein to astrocytes in the white matter of human cerebrum. No specific staining of neurones or of other glial elements was detected.
Brain type creatine kinase-BB (CPK-BB) was measured by radioimmunoassay in the serum of 113 women with breast cancer and 354 female controls. 80% of women with metastatic breast cancer had levels above 3 ng/ml (control range 0.5--3.7 ng/ml); the highest level was 23 ng/ml. 60% of women with local disease but no evidence of distant metastases showed levels above 3 ng/ml, the highest being 9.0 ng/ml. Of women who had presented with stage I, II, or III disease and postoperatively had no evidence of persistent disease 30% had levels above 3 ng/ml. Serial measurements in 31 patients indicated that the serum CPK-BB correlated with clinical response to treatment.
Brain-type creatine kinase isoenzymes (CK-BB) was measured by radioimmunoassay in the serum of 54 patients with head injuries. CK-BB was not detectable in 476 out of 1006 controls, the remaining 530 normal samples containing a mean of 1.5 +/- SD0.75 microgram/l. The mean CK-BB concentrations in patients with mild, moderate, and fatal head injuries were all significantly higher than the control value (p < 0.01 in each instance). Patients with serious head injury had serum concentrations many times the normal value, in two cases within 30 minutes after impact. Fatally injured patients continued to have high serum concentrations several days after injury. In less serious cases values approached normal within two or three days. Every patient with evidence of cerebral laceration, bruising, or swelling had a serum CK-BB concentration above normal. Raised concentrations were found in 14 out of 22 patients with concussion only. The serum CK-BB concentration appears to be a sensitive index of brain damage and may prove useful in the management and follow-up of head-injured patients.
Nervous system-specific 14-3-2 protein is an acidic enolase (EC 4.2.1.11) isoenzyme which is found in high concentrations in nervous tissue and which is also considered to be a molecular marker for amine precursor uptake and decarboxylation (APUD) cells. A radioimmunoassay for this protein shows levels of immunoreactivity of 10 ng/ml and 5 ng/ml in human cerebrospinal fluid and serum respectively. The immunoreactivity in concentrated cerebrospinal fluid dilutes out in parallel with the standard curve in the assay, has a similar molecular weight to the immunoreactivity in soluble protein extracts of whole human brain, and on polyacrylamide gel electrophoresis migrates as two forms representing the homodimer and probably the heterodimer species of the enzyme. The protein also occurs at relatively high levels in erythrocytes and an approximately 1% haemolysis would be sufficient to account for the normal serum level of immunoreactivity. Clinical measurements of 14-3-2 protein in neurological disorders or APUD-system disease could be potentially useful in cerebrospinal fluid but are unlikely to be of value in serum.