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Biomedical subjects

J Chayen

Publications and source records attributed to J Chayen.

At least 73 records · Page 4Linked to original sources

Synthesis of arachidonate cyclo-oxygenase products by rheumatoid and nonrheumatoid synovial lining in nonproliferative organ culture.

Specimens of human rheumatoid and nonrheumatoid synovial lining were maintained in nonproliferative organ culture for 20 hours. The culture fluids were then assayed for prostaglandin E(2) (PGE(2)), thromboxane B(2) (TXB(2)), and 6-keto-prostaglandin F(1alpha) (6-keto-PGF(1alpha)) by specific radioimmunoassay. The presence of each of these substances was confirmed by gas chromatography and mass spectrometry. Rheumatoid tissue produced significantly more of each cyclo-oxygenase product than nonrheumatoid tissue.

6-Ketoprostaglandin F1 alpha↗

A modified tetrazolium reaction for identifying malignant cells from gastric and colonic cancer.

When non-malignant cells were reacted for glucose-6-phosphate dehydrogenase activity, with neotetrazolium chloride as the indicator of the activity, oxygen competed with the neotetrazolium and nullified the reaction. In contrast, about 30% of the activity was retained in malignant cells, in sections and in smears, from cancer of the stomach or colon. This could provide the basis of a qualitative (black-or-white) functional test for distinguishing malignant cells in these conditions.

Colonic Neoplasms↗

Changes in crystal size and orientation of acidic glycosaminoglycans at the fracture site in fractured necks of femur.

The aim of this study was to try to elucidate the increased susceptibility of the neck of femur to fracture. Quantitative polarised light microscopy has been applied to fresh, undecalcified sections of samples of bone taken from the site of fracture, in specimens taken at operation from patients with fractures of the femoral neck or osteoarthritic femoral heads or from the equivalent site from otherwise normal subjects at necropsy. In all 21 specimens of fractured necks of femur, but in none of the other specimens, relatively large crystals (up to 2.5 X 0.5 micrometres) were found close to the site of fracture; the properties of these crystals were compatible with their being apatite. Measurement of the natural birefringence of the collagen showed no difference in the orientation of the collagen in all three types of specimen. However, the orientation of acidic glycosaminoglycans, measured by the birefringence of alcian blue bound to these moieties, was 45 per cent lower in the specimens from fractured necks of femur than in the other specimens, even though the total content of acidic glycosaminoglycans was unchanged. Although the decreased orientation was most marked close to the site of fracture, it was still apparent 15 millimetres from that site. These changes were unlikely to be simply the sequelae of fracture since they were not found in traumatic fractures of other bones. Thus it is conceivable that changes in the orientation of the ground substance allow formation of relatively large crystals of apatite and that such crystals, in the microcrystalline mass of apatite, are the cause of the increased fragility of such bones.

Aged↗

A quantitative cytochemical method for the measurement of beta-hydroxyacyl CoA dehydrogenase activity in rat heart muscle.

Although cytochemical methods exist for measuring dehydrogenases that act on substrates involved in the production of chemical energy from sugars, virtually no methods exist for measuring the dehydrogenases that act on fatty acids. Yet the oxidation of fatty acids accounts for over 60% of the oxidative activity of cardiac muscle. Consequently a new quantitative cytochemical method, based on a new substrate (DL-S-beta-hydroxybutyryl-N-acetyl cysteamine), has been developed for measuring the activity of hydroxy-acyl coenzyme A dehydrogenase, which is the penultimate step of the beta-oxidation of fatty acids to acetyl-coenzyme A that is used in the Krebs' cycle. Menadione or phenazine methosulphate is used as the intermediate hydrogen-acceptor, with neotetrazolium chloride as the final acceptor. The medium contains nitroprusside, ostensibly to react with any cysteamine liberated by hydrolysis of the substrate. As a control, cysteamine is substituted for the substrate. The concentrations of reactants have been optimized for cardiac muscle; the reaction is linear with thickness of the sections and with time of reaction from 15 to 60 min.

3-Hydroxyacyl CoA Dehydrogenases↗

The involvement of the pentose shunt in thyroid metabolism after stimulation with TSH or with immunoglobulins from patients with thyroid disease. 1. The generation of NADPH in relation to stimulation of thyroid growth.

It has been shown previously that both thyrotrophin (TSH), and also immunoglobulins (Ig) derived from patients with goitrous Graves' disease, stimulate DNA-synthesis in guinea-pig thyroid tissue maintained in vitro. Here we describe the use of the same in vitro system and methods of quantitative cytochemistry to test the effect of these substances on the generation of NADPH, which is another indicator of the potential for growth. As could be predicted by its trophic action, TSH stimulated the generation of NADPH by glucose 6-phosphate dehydrogenase. The Ig-fraction from normal subjects depressed this activity. The Ig-fraction from Graves' disease patients with goitres stimulated the generation of NADPH, whereas the Ig from patients with Graves' disease but with minimal enlargement of the thyroid gland behaved like normal Ig. A similar lack of stimulation was found with Ig from patients with Pendred's syndrome, other dyshormonogenetic goitres, and autonomous single adenomas. In all specimens tested, there was good correlation between the amount of DNA-synthesis, measured by Feulgen cytophotometry, and the activity of glucose 6-phosphate dehydrogenase activity that generated NADPH. These results support the concept that there is a distinct type of autoantibody that influences thyroid growth.

Animals↗

The involvement of the pentose shunt in thyroid metabolism after stimulation with TSH or with immunoglobulins from patients with thyroid disease. II. The reoxidation of NADPH and stimulation of hormone synthesis.

Reducing equivalents derived from the tissue re-oxidation of NADPH (NADPH-diaphorase) have been implicated in the peroxidation that is involved in the organification of iodine in the production of thyroid hormones. Immunoglobulin (Ig) fractions from patients with thyroid diseases and from normal controls, in a standard dose of 125 micrograms/ml and 0.3 microunits/ml thyrotrophin (TSH) were incubated with segments of guinea-pig thyroid gland maintained in vitro. A quantitative cytochemical study was made on how these fractions influenced the enzyme activity. A good correlation was found between the ability of such Ig fractions to stimulate the NADPH-diaphorase activity and (1) the degree of hyperthyroidism in the patients and (2) the amount of T3 secreted by the thyroid segments in vitro.

Animals↗

Cell division in the synovial lining in experimental allergic arthritis: proliferation of cells during the development of chronic arthritis.

The synovial tissue in experimentally induced immune arthritis induced in the rabbit has been used as a model of rheumatoid arthritis to determine which cells may contribute to the growth of this tissue. Tissue from the challenged and from the unchallenged knee joints was taken, after the intra-articular injection of a small amount of tritiated thymidine, from rabbits up to 3 months after the arthritis was induced. DNA synthesis, as a measure of cell proliferative activity, was assessed firstly by measuring the labelling index in autoradiographs of sections of such tissue, and secondly by the DNA synthetic index obtained by Feulgen cytophotometry. These measurements were made separately on synoviocytes, on the structural cells of the stroma, on the cells lining the small blood vessels, and on the infiltrating inflammatory cells. The DNA synthetic activity of the synoviocytes, and of the stromal noninflammatory cells, was maximal between 3 and 7 days after challenge. The activity in the synoviocytes, in particular, remained raised for up to 84 days after the challenge. Thus these cells appear to be capable of contributing to the hyperplasia, but the contribution of other cells, deeper in the stroma, cannot be excluded.

Animals↗

Circulating levels of biologically active parathyroid hormone in rheumatic diseases.

There has been doubt as to whether elevated levels of parathyroid hormone, reported previously by radioimmunoassay, reflect increased concentrations of the biologically active hormone. The application of a recently developed, highly sensitive bioassay has shown considerable disparity between bioactivity and immunoreactivity in 5 rheumatic conditions and in normal subjects. Six patients with chondrocalcinosis had elevated levels; 3 of these did not have hypercalcaemia or any obvious cause other than possible subclinical hyperparathyroidism. One patient, assayed during an acute episode, had an elevated concentration of the hormone which reverted to normal when she was asymptomatic. Most patients with osteoarthrosis (13 our of 15) had low normal levels; 2 showed unexplained slightly elevated concentrations. Of 6 patients with haemochromatosis 3 had elevated levels, though this may have been related to the associated presence of diabetes mellitus. A third of patients with ankylosing spondylitis (10 out of 30) showed elevated parathyroid hormone levels but without hypercalcaemia. A number of spondylitic patients also showed anomalous results in this assay, possibly due to the presence of an antagonist. This would be consistent with the absence of clinical or biochemical evidence of hyperparathyroidism.

Adult↗

Feulgen-hydrolysis profiles in cells exfoliated from the cervix uteri: a potential aid in the diagnosis of malignancy.

By varying the time of hydrolysis for the Feulgen reaction, done under conditions that protect the backbone of the DNA, it is possible to distinguish three species of DNA that are characterised by their lability to acid hydrolysis. The most labile DNA was found, in greatest proportions, in malignant cells; this may be helpful in diagnostic cytology. The fact that the cytologically normal cells, in grade V smears, also show this labile DNA may well facilitate cytological screening even in those smears that contain very few neoplastic cells.

Cervix Uteri↗

Inhibition of cytochemical bioactivity of parathyroid hormone by plasma in pseudohypoparathyroidism type I.

Despite the high circulating levels of immunoreactive PTH in patients with pseudohypoparathyroidism type I (PSPI) the levels of bioactive PTH (bioPTH) have been found to be close to the normal range. To elucidate this dissociation, we have studied the recovery of the biological activity of bovine PTH added to the plasma of patients with either PSPI, or with hypoparathyroidism (PTX), primary hyperparathyroidism (HPT) or of normal subjects. In PSPI (n = 10) the recovery of biological activity was 5.6% +/- 3.6 (mean +/- SEM) whereas in PTX (n = 7), in HPT (n = 4) and in normal subjects (n = 8) it was 79% +/- 5, 75% +/- 9 and 68% +/- 4, respectively. In another PSPI patient, who had undergone total parathyroidectomy, bioPTH was undetectable but the recovery from the plasma of added PTH was 84%. Thus we have found inhibition of PTH bioactivity by plasma of PSPI patients which was absent after parathyroidectomy.

Animals↗

Experimental allergic arthritis in the rabbit: alterations in the cellularity and the rate of cellular proliferation in the synovial linings of the challenged joints of rabbits immunized with antigen in Freund's incomplete adjuvant.

Rabbits have been immunized with ovalbumin in Freund's incomplete adjuvant (FIA) followed by the intra-articular injection of ovalbumin, in order to follow the development of inflammation in the synovial lining. The kinetics of cell proliferation have been investigated using tritiated thymidine (3HTdR) autoradiography and Feulgen cytophotometry. Unexpectedly, marked histological changes were found in the synovium, with hyperplasia of synoviocytes, and of the connective-tissue cells of the subintima, being seen as early as 3 days after challenge. Large numbers of inflammatory cells, including many plasma cells, were found in the synovium at between 5 and 11 days. Labelling of synoviocytes and connective-tissue cells reached a maximum at 3 days and declined thereafter, reaching normal levels at 14 days. Three weeks after challenge the synovium was normal in appearance.

Animals↗

Thyroid growth-blocking antibodies in primary myxoedema.

Adult primary myxoedema is usually due to an autoimmune thryoiditis characterized by progressive shrinking of the thyroid gland, loss of epithelium, dense infiltration by sensitized lymphocytes and plasma cells with final replacement of the gland by a fibrous scar. Antibodies directed against thyroglobulin (TgHA) and microsomal antibodies (McHA), detectable years before the onset of hormonal failure, and cell-mediated immune mechanisms contribute to the pathogenesis. When 90% of the gland is destroyed, the secretion of thyroid hormones (T3/T4) falls below normal needs and pituitary thyrotrophs react by producing over 100 times the normal amount of thyroid-stimulating hormone (TSH) which should normally lead to regrowth of the gland as happens in goitrous Hashimoto thyroiditis. The inability of the thyroid in primary myxoedema to respond to the trophic action of TSH suggested that blocking antibodies exist in this disease which compete with TSH for its receptors. The trophic effect of TSH can be assayed in vitro by measuring DNA-synthesis by Feulgen cytophotometry; elevated pentose-shunt oxidative activity correlates with DNA synthesis. Here we report that immunoglobulins from patients with adult, primary myxoedema block the trophic effect of TSH, as assessed by this in vitro system. This is in marked contrast to the growth stimulation induced by immunoglobulins from patients with thyrotoxic Graves' disease with prominent goitres.

Autoantibodies↗

The assessment of DNA-synthetic activity.

A method is described by which a numerical value can be assigned to the amount of DNA-synthesis shown graphically by population-histograms obtained by Feulgen cytophotometry. The index appeared to give a reasonable measure of DNA-synthetic activity in populations having very low, moderate and high mitotic indices and closely followed labelling indices obtained by autoradiography. Thus the advantage of a numerical value is that the DNA-synthetic activities in different populations can be compared for statistical analysis.

Animals↗

The use of a hidden metal-capture reagent for the measurement of Na+--K+-ATPase activity: a new concept in cytochemistry.

The original lead-trapping method for demonstrating Na+--K+-ATPase activity was discredited because of the effect that lead ions can have on the substrate and on the enzyme. Current methods, that measure this activity by the related K+-dependent phosphatase activity, do not appear to measure activity that is known, from microchemistry, to occur in proximal convoluted tubules. The disadvantages of using lead appear to have been overcome by the use of a new reagent in which the lead is complexed with ammonium citrate ions; phosphate, liberated enzymatically, successfully competes with these ions. The activities of total ATPase and of the ouabain sensitive Na+--K+-ATPase have been measured in three regions of the nephron in the guinea-pig and in the rat. The relative activities found, by this method, in the different regions of the latter, appear to be comparable with results found by others, using microchemical methods applied to isolated regions of the nephron.

Animals↗