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Biomedical subjects

E Baron

Publications and source records attributed to E Baron.

At least 19 recordsLinked to original sources

Amplification and expression of the epidermal growth factor receptor gene during the progression of neuroepithelial tumours in vivo.

The epidermal growth factor receptor (EGFR) gene is homologous to the oncogene c-erbB. The occurrence of amplification and rearrangements at the EGFR gene locus is associated with malignancy in neuroepithelial tumours. Sixteen neuroepithelial tumours from eight patients with recurrence of their neoplasms were analysed for changes at the EGFR gene locus and for expression of EGFR. Ten tumours from five patients lacked changes at the EGFR gene locus. Three of eight individuals showed EGFR gene amplifications in both tumours with a higher grade of amplification in the second tumour. In addition to amplification, a rearrangement was found in both tumours of the first patient. In the second case an amplification of chromosome-7-specific c-met sequences was found in the regrown tumour, suggesting that a polysomy 7 was at least partly responsible for the higher copy number of the EGFR sequences. In both tumours of the third patient with EGFR gene amplification different alleles were amplified. In contrast to the findings at the DNA level the EGFR expression, analysed by immunohistochemical techniques, showed a more heterogeneous pattern after tumour progression.

Adult

Clinical importance of Bilophila wadsworthia.

Bilophila wadsworthia is an anaerobic, gram-negative, asaccharolytic, urease-positive, bile-resistant, catalase-positive bacillus, originally recovered from infections in patients with gangrenous and perforated appendicitis. Additional isolations from clinical specimens, including pleural fluid, joint fluid, blood and pus from a scrotal abscess, mandibular osteomyelitis and axillary hidradenitis suppurativa are described here. Bilophila is found as normal flora in feces and, occasionally, in saliva and in the vagina. Isolates from humans are usually beta-lactamase positive and therefore resistant to certain beta-lactam antibiotics. Two percent of strains are also resistant to clindamycin.

Adult

Changes in stimulating and blocking TSH receptor antibodies in a patient undergoing three cycles of transition from hypo to hyper-thyroidism and back to hypothyroidism.

We report a patient who underwent, over a mere 3-year period, three successive cycles of oscillation from hypo to hyper-thyroidism and back to hypothyroidism. This unusual sequence of events originated in a rare passage of primary hypothyroidism to hyper-thyroidism. The hyperthyroidism seemed typical of the autoimmune subgroup of toxic multinodular goitre. Stimulating and blocking TSH receptor antibody activities were measured (by cAMP functional bioassays using cultured human thyrocytes) during the course of the fluctuating phases of hypo and hyper-thyroidism. Measurement of such antibody activities revealed the coexistence of both stimulatory and blocking types of antibody in several serum samples from the patient. Throughout the whole course of alterations in thyroid function, thyroid stimulating antibodies were present. This was not the case with thyrotrophin receptor antibodies exhibiting TSH antagonist activity which seemed to appear and disappear. Monitoring such activity indicated that the emergence of blocking antibody seems to herald the onset of hypothyroidism.

Autoantibodies

Divergent effects of thyroid HLA-antigens on T and natural killer (NK) cell cytotoxicity.

We have used non-autoimmune non-neoplastic human thyroid cells to explore the role of surface class I and DR antigens on these cells' sensitivity towards T and Natural Killer (NK) cell cytotoxicity. Non-treated thyrocytes expressed class I but no DR antigens. Following incubation with gamma-interferon (gamma-IFN) class I antigens were markedly elevated and DR expression was induced. Whereas non-treated thyrocytes were minimally lysed by sensitized T cells, they served as appropriate targets for NK cells. Following incubation with gamma-IFN, the thyroid cells became highly sensitive to T cell lysis, with no significant reduction in their vulnerability to NK cell killing. The addition of monoclonal anti class I or DR antigens, or brief acid treatment which specifically eliminates class I molecules, inhibited T cell cytotoxicity but enhanced the sensitivity to lysis by NK cells. Thus, the presence of HLA antigens on the same thyroid cells have an opposite effect on two major cytotoxi mechanisms. Our findings are relevant within the context of recent suggestions of intervening with target HLA antigens for the management of autoimmune and malignant diseases.

Cytotoxicity, Immunologic

Telenzepine inhibits electrically-stimulated, acetylcholine plus histamine-mediated acid secretion in the mouse isolated stomach by blockade of M1 muscarine receptors.

1. The muscarine receptor mediating electrically-stimulated acid secretion in the mouse isolated stomach was characterized using a variety of muscarine receptor antagonists confirming the M1 nature of the antagonist effect of telenzepine. 2. Field stimulation (7 V, 10 Hz, 0.5 ms) resulted in a plateau acid secretion over at least 90 min which was completely blocked by either 1 mumol/l TTX or H2 receptor antagonists (100 mumol/l cimetidine or 10 mumol/l lupitidine). Ranitidine, which is known to potently inhibit mucosal acetylcholine esterase, was ineffective. Compound 48/80 at 100 mumol/l, which depletes mucosal histamine stores, initially mimicked electrical stimulation but subsequently prevented it from inducing acid secretion. 3. 10 muscarine receptor antagonists with differing relative affinities for M1, M2 and M3 receptors were introduced at 1 mumol/l to inhibit electrically-stimulated acid secretion. The percentages inhibition were plotted against binding affinities of the antagonists at either M1, M2 or M3 binding sites. A statistically significant correlation between functional and binding data was detected only when based on M1 affinities. 4. It is concluded that field stimulation, which probably mimicks vagal drive, results in muscarinic M1 receptor activation on paracrine cells to release histamine. Histamine then stimulates parietal cells to secrete acid. Hence, according to the present and our previous data, telenzepine inhibits acid secretion under these conditions by blocking M1 receptors at least partially located on histamine-releasing paracrine cells.

Acetylcholine

Early and late lesion-to-non-lesion ratio of thallium-201-chloride uptake in the evaluation of "cold" thyroid nodules.

Forty-nine consecutive patients with "cold" thyroid nodules were studied using early and late visual and semiquantitative measurements of 201Tl uptake in the nodule to differentiate benign from malignant nodules. The visual method compared 201Tl uptake in the nodule to the normal thyroid tissue. The semiquantitative method used a lesion-to-non-lesion (L/N) ratio of the same areas. Both measurements were carried out early (15 min) and late (3 hr) following 201Tl injection. The reproducibility of the method for the early and late measurements was tested for intraobserver and interobserver variability as well as for repeatability coefficients. The visual method resulted in 43% sensitivity and 79% specificity for the detection of malignant nodules. The L/N method showed that an early threshold of 1.55 chosen by receiver characteristic analysis had a sensitivity of 57% and a specificity of 86%, while the late ratio of 0.99 had a sensitivity of 100% and a specificity of 62%. It is concluded that a L/N 201Tl uptake method performed 3 hr following 201Tl injection is superior to a visual scoring method as well as to the early L/N 201Tl uptake in detecting malignant thyroid nodules.

Adenoma

Long-term pergolide treatment of hyperprolactinemic patients previously unsuccessfully treated with dopaminergic drugs.

Nine hyperprolactinemic patients (seven women aged 25-57 and two men aged 23-51 years) were treated with the dopamine agonist drug, pergolide mesylate, for a period ranging from 4 months to 5 years. All patients had previously received different dopamine agonist drugs; additionally two of the group had undergone surgery and radiation therapy. Group 1 comprised five patients who were given pergolide in an attempt to avoid the marked side effects of bromocriptine and lisuride that had been given earlier. Group 2 comprised four patients treated with pergolide following failure to respond to other dopamine agonist drugs (bromocriptine, lisuride). In group 1, pergolide, at a dose of 25-50 micrograms/day, caused a decrease in the prolactin level to the normal value, restored menstruation, and very few side effects were noted. In group 2, pergolide was administered in a gradually increasing dose, the patients receiving a maximum of 200 micrograms/day. At this dose, there was a significant decrease in the prolactin level in two patients compared with only a slight decrease in the remaining two patients. This study illustrates the efficacy of pergolide in treating hyperprolactinemic patients unable to tolerate other dopaminergic agents (bromocriptine, lisuride).

Adult

Characterization of the muscarine receptor type on paracrine cells activated by McN-A-343 in the mouse isolated stomach.

An attempt was made to characterize the muscarine receptor type(s) involved in acid secretion in the mouse isolated stomach when stimulated by the muscarinic agonist McN-A-343. A series of 8 muscarinic antagonists was used with preference for M1 receptors (telenzepine and pirenzepine), M1 and M2 receptors (secoverine), M2 receptors (AF-DX 116 and himbacine) and M1 and M3 receptors (p-F-HHSiD and HHSiD). BTM-1086 was used as a high affinity antagonist at the M1 receptor however with little selectivity. Receptor type preferences were determined in binding experiments with [3H]telenzepine in cortical membranes (M1) and [3H]N-methylscopolamine in atrial (M2) or salivary gland (M3) membranes, derived from guinea pigs. No antagonist with M3 preference could be identified in the binding studies. A fixed antagonist concentration of 1 mumol/l was used to antagonize acid secretion stimulated by 10 mumol/l McN-A-343. By plotting the percentage inhibition obtained in the functional test against the Ki values determined in binding experiments for each antagonist at M1, M2 and M3 binding sites, an affinity-inhibition curve could only be constructed when based on the antagonist affinities to the M1 receptor. No statistically significant fit was found using antagonist affinities to the M2 or M3 receptor. Thus, in accordance with the presumed M1 selectivity of the agonist McN-A-343, the rank order of potencies of different antagonists point to the M1 nature of the muscarine receptor which stimulates acid secretion in the mouse isolated stomach upon activation by McN-A-343.(ABSTRACT TRUNCATED AT 250 WORDS)

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy

From cloning to a commercial realization: human alpha interferon.

Recent applications of recombinant DNA techniques have enabled the cloning of several interesting human genes, leading to the production of rare biologicals in abundant quantities. We review here the discovery, early characterization, cloning, and expression of Interferon Alfa-2B (IFN alpha-2b or Intron A) as a therapeutic at Schering-Plough Research. IFN alpha-2a is marketed by Hoffman LaRoche under the trade name Roferon. The studies on the expression, purification, biology, and clinical aspects of this interferon offer a plethora of information on one of the earliest recombinant DNA based drugs to reach the market place.

Amino Acid Sequence

Class II HLA-DR antigens on non-autoimmune human thyroid cells stimulate autologous T cells with high suppressor activity.

Human autoimmune thyroid cells "spontaneously" express MHC-class II antigens. These antigens have been assumed to trigger thyroid-specific helper T cell clones, leading in turn to the expansion of thyroid autoantibody-secreting B cells. Thyroid cells derived from non-autoimmune subjects do not express MHC-class antigens, but these can be readily induced with gamma-interferon. We have addressed the issue of whether it is sufficient for normal thyroid cells to bear class II antigens in order to trigger autologous T cells. We found that non-autoimmune thyrocytes expressing DR antigens fail to stimulate autologous resting T cells. However, proliferative activity and interleukin-2 secretion were observed when fresh T cells were first triggered by autologous non-T cells and then incubated with thyrocytes. More CD8 than CD4 cells proliferated in the T:thyrocyte cultures, but CD4 cells were necessary for the proliferation and interleukin-2 secretion. Addition of antibodies to thyroglobulin or to DR antigens inhibited T cell proliferation and interleukin-2 secretion, thus pointing to T cell recognition of both thyroid-specific and DR antigens. Evaluation of the function of the thyroid stimulated T cells revealed very potent suppressor but negligible helper and cytotoxic activities. It would seem, therefore, that DR-restricted T cell activation by autologous antigen on non-autoimmune thyroid cells does occur, but since it results in enhanced suppression, its nature seems protective, thus leading to maintenance of immunological self-tolerance.

Antigen-Presenting Cells

Stimulation by McN-A-343 and blockade by telenzepine of acid secretion in the mouse isolated stomach at histamine-liberating cells.

(1) In the lumen-perfused mouse stomach in vitro, potential sites of gastric antisecretory action of the muscarine M1-receptor antagonist telenzepine were investigated. Acid secretion was stimulated by the muscarinic agonist McN-A-343 (1-1000 mumol/l). Neither basal nor McN-A-343-stimulated acid secretion was affected by 1 mumol/l TTX indicating that neuronal structures were probably not involved. (2) Acid secretion stimulated by 10 mumol/l McN-A-343 was inhibited by telenzepine (0.1-1.4 mumol/l) and cimetidine (10-140 mumol/l). Neither of the antagonists affected basal acid secretion. TTX had no inhibitory influence on the antagonist effect of telenzepine and cimetidine. (3) Compound 48/80 (100 mumol/l), which depletes histamine stores, initially mimicked but subsequently prevented the effect of McN-A-343. Prenylamine (50 mumol/l), which prevents histamine release, also abolished the secretagogue effect of subsequently administered McN-A-343. (4) Up to concentrations greater than 100 mumol/l, McN-A-343 did not stimulate acid production in rabbit isolated fundic glands and guinea-pig isolated parietal cells. Thus, parietal cells are not directly stimulated by McN-A-343. (5) Based on the site of action of the agonist McN-A-343 in the mouse isolated stomach and its failure to stimulate parietal cells from different species directly, it is concluded that telenzepine blocks, in the mouse isolated stomach, muscarine receptors located on paracrine cells to reduce endogenous histamine release.

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy

Thyrotrophin receptor blocking antibodies: incidence, characterization and in-vitro synthesis.

The prevalence and characteristics of TSH receptor blocking activity were examined in patients with different thyroid disorders. Studies were also performed on the in-vitro synthesis and immunoregulation of the blocking antibody. Blocking activity was tested by measuring the inhibition of TSH-stimulated cAMP production of cultured human thyroid cells by patient immunoglobulin (Ig) preparations. The following patients were investigated (number of cases in parentheses): Hashimoto's thyroiditis (33); primary myxoedema (17); euthyroid ophthalmopathy (8); active Graves' disease (19); cold nodule (5); non-toxic goitre (14); toxic adenoma (8); toxic multinodular goitre (9) and 22 normal controls. TSH receptor blocking activity was only detected in primary hypothyroidism with the following characteristics: (i) Such activity was present in only 16% of the patients (both goitrous, i.e. Hashimoto's thyroiditis, and non-goitrous, i.e. primary myxoedema), and in three patients with previously active Graves' hyperthyroidism who had become hypothyroid. (ii) Blocking activity seems to be associated with the Ig fraction of serum as indicated by protein A adsorption. (iii) The block-positive samples did not bind 125I-TSH, which seems to rule out an antibody directed against TSH. (iv) The specificity of the blocking activity seems to be directed towards the TSH-(thyroid stimulating immunoglobulin, TSI) receptor-mediated cAMP response since no inhibition of prostaglandin E1-stimulated cAMP production was found. Moreover, all cases in which an inhibitory effect was demonstrated towards TSH also exhibited blocking of TSI-stimulated cAMP, with a high correlation between the degree of inhibition of the TSH to that of the TSI response (r = 0.89, P less than 0.001, n = 11). The blocking activity may contribute to the pathogenesis of some cases of primary hypothyroid autoimmune thyroiditis, both goitrous and non-goitrous, as well as in the evolution of hyper- to hypothyroidism. By culturing peripheral blood lymphocytes, as well as B/T lymphocyte co-cultures isolated from three patients with blocking activity present in serum, the in-vitro synthesis of the blocking antibody was demonstrated for the first time. Moreover, in-vitro secretion of the antibody by patients B lymphocytes, as well as T cell regulation of autoantibody production, were also shown.

Adult

L-Thyroxine-induced leukopenia in a patient with Hashimoto's disease: involvement of suppressor-cytotoxic T cells.

A study of in vitro reactivity to L-T4, as assessed by peripheral blood lymphocyte transformation, was carried out in a patient with Hashimoto's disease who developed leukopenia during treatment with L-T4. L-T4 stimulated lymphocyte transformation in this patient. The proliferating cells were of T lineage and further typing demonstrated that they bore the OKT8 antigen. Cell-mediated immune reactions mediated by OKT8+T cells might have caused the leukopenia, although other mechanisms involving cytotoxic antibodies or antibody-dependent cell-mediated cytotoxicity cannot be excluded.

Female