Unlike thyrotropin, thyroid-stimulating antibodies do not activate phospholipase C in human thyroid slices.

The effects of thyroid-stimulating antibodies (TSAb) and of thyrotropin (TSH) were compared, on the generation of cyclic AMP and inositol phosphates (InsP), in human thyroid slices incubated in vitro, and on the Rapoport cyclic AMP bioassay. The TSAb positive sera were obtained from 19 patients with Graves' disease. In 14 experiments with the slices system, TSH significantly increased cyclic AMP accumulation (TSH, 0.03-10 mU/ml) as well as the cyclic AMP-independent inositol trisphosphate (InsP3) generation (TSH, 1-10 mU/ml). In the same 14 experiments, TSAb (0.10-28 mg/ml) enhanced cyclic AMP intracellular levels as expected while they did not induce any InsP accumulation. Even when TSAb increased cyclic AMP levels to the same or higher values as those obtained with TSH concentrations allowing InsP3 generation. TSAb were still unable to activate the phosphatidylinositol-Ca2+ cascade. The patterns of the response curves of TSAb and TSH on cyclic AMP accumulation were different, suggesting that different mechanisms may be involved. In addition, unlike TSH, TSAb were not able to stimulate H2O2 generation, which in human tissue mainly depends on the activation of the phosphatidylinositol-Ca2+ cascade. Immunoglobulins from six additional Graves' patients lacking measurable cyclic AMP-stimulating activity in both slices and cells systems did not activate phospholipase C either. In conclusion, our results show that TSAb do not share all the metabolic actions of TSH on human thyroid tissue. The data provide support for the concept that the pathogenesis of Graves' disease can be fully accounted for by the ability of TSAb to stimulate adenylate cyclase. This work also confirms that TSH activates the cyclic AMP and the phosphatidylinositol cascade by independent pathways in the human thyroid.

A randomized study comparing a high and a standard dose of cisplatin in combination with etoposide in the treatment of advanced non-small-cell lung carcinoma.

We conducted a randomized trial comparing a high (120 mg/m2 day 1) v a standard (60 mg/m2 day 1) dose of cisplatin in combination with etoposide (120 mg/m2 days 3, 5, and 7) in advanced non-small-cell lung carcinoma (NSCLC). Two hundred forty-one patients were evaluable for survival and 207 for response. We obtained a 25% objective response rate in the standard-dose arm and 29% in the high-dose arm; this difference was not statistically significant. There was no significant improvement in the overall survival or survival of responders with the high-dose regimen. However, toxicity (mainly myelosuppression) was significantly increased in the patients receiving the higher dose of cisplatin. An analysis of prognostic factors showed that disease progression, loss of body weight, performance status, and prior therapy were predictive parameters of survival.

Hormonal secretion by hyperactive thyroid cells is not secondary to apical phagocytosis.

Hyperactive dog thyroids were prepared by repeated TSH stimulation in vivo. Butanol-extractable 125I (BE125I) release in vitro from slices and hormonal secretion in vivo in the thyroid vein was enhanced. Apical pseudopods and colloid droplets were very infrequent in such hyperactive thyroids. Moreover, BE125I release was insensitive to cytochalasin B, to inhibitors of microtubules, and to metabolic inhibitors, and decreased only with temperature. Our data demonstrate that hormonal secretion by hyperactive thyroids is not secondary to apical phagocytosis (i.e. macropinocytosis). Micropinocytosis or intraluminal hydrolysis of thyroglobulin are suggested as first step of the secretory process.

Regulation and metabolic role of phospholipase D activity in human thyroid and cultured dog thyrocytes.

The actions of TSH, ATP, the ionophore A23187, the endoplasmic reticulum Ca(2+)-ATPase inhibitor thapsigargin, and phorbol dibutyrate (PDBu) on 3H-cytidine-monophosphate phosphatidic acid (3H-CMP-PA) accumulation were studied in human thyroid slices to evaluate PA generation and inositol recycling towards phosphatidyl-inositol synthesis. The effects of the same agonists also were measured on phosphatidylbutanol (PtdBut) generation in 3H-palmitate or 3H-myristate prelabeled slices to assess the activity of phospholipase D (PLD). The phospholipid target of this PLD was determined on 3H-choline prelabeled human thyroid slices by measuring 3H-choline release in incubation medium and slices and 3H-choline incorporation in phospholipids. TSH (10 U/L) stimulated 3H-CMP-PA accumulation in an LiCl-and propranolol-insensitive way, as well as 2H-fatty acids incorporation into PA, diacylglycerol, and phosphatidylcholine (PtdCho) with on evidence of dose-dependent effects and had no detectable action on PLD activity. The effects of TSH were not reproduced by Bu2cAMP or forskolin. Thapsigargin and A23187 both increased CMP-PA accumulation and PtdBut generation, whereas ATP only stimulated PLD activity. The phorbol ester PDBu (5 x 10(-7) mol/L) increased PtdBut formation and 3-H-fatty acid incorporation into PtdCho, but had no effect on CMP-PA generation. Staurosporine (STSP) (5 x 10(-6) mol/L), a nonspecific inhibitor of protein kinase C, unexpectedly reproduced the effects of PDBu. The increase of 3H-choline in slices' supernatant and the decrease of 3H-choline-labeled PtdCho induced by PDBu, ATP, thapsigargin, and STSP indicate that the activated PLD hydrolyzed PtdCho. We suggest that the PA generation induced by PLD stimulation could contribute to the stimulated H2O2 formation and iodide organification observed with the agonists inducing PtdBut accumulation. Indeed, Bu2cAMP and forskolin, known to decrease iodide organification in human thyroid, inhibited the PLD stimulation induced by ATP and PDBu. In cultured dog thyrocytes, phorbol esters, and STSP induced DNA synthesis and dedifferentiation, whereas thapsigargin inhibited TSH-induced growth and killed phorbol esters stimulated cells, suggesting a positive role of PLD stimulation towards dedifferentiated growth and of simultaneously raised [Ca2+)i and stimulated protein kinase C-PLD towards growth arrest and cellular death.

Pathogenesis of autonomous thyroid nodules: in vitro study of iodine and adenosine 3',5'-monophosphate metabolism.

The in vitro characteristics of iodide and cAMP metabolism have been compared in tissues from autonomously functioning thyroid nodules and their quiescent counterpart to test the hypothesis that autonomy may result from constitutive activation of the tissue's TSH, cAMP, and protein phosphorylation regulatory axis, as in vivo nodular tissue took up more iodide. This effect was entirely due to increased transport capacity, the affinity of iodide transport, and the fractional binding of iodide to protein remaining unchanged. However, at high concentrations total iodide binding to protein was similar in quiescent and nodular tissue. In both tissues, this metabolic step was enhanced by phorbol esters and the ionophore A23187. As evaluated by autoradiography of two-dimensional gel protein electrophoregrams, no differences in the patterns of protein synthesis or phosphorylation between quiescent and nodular tissue were found. Basal cAMP levels were similar in quiescent and nodular tissue. The cAMP response to TSH was lower in nodular tissue, with no change in sensitivity or kinetics; both tissues responded to forskolin. No systematic suppression of iodide inhibition or abnormal responses to other hormones or neurotransmitters were found. Three proteins (24K-1, 24K-2, and 26K) were phosphorylated only in the presence of TSH or forskolin in both quiescent and nodular tissue. One protein substrate (20K) was phosphorylated in the presence of TSH in the quiescent, but not in the nodular, tissue. In conclusion, 1) slices from autonomous thyroid nodules reproduce the in vivo characteristics of the lesion and are, therefore, a suitable in vitro experimental model for biochemical studies; 2) taken together with data from transplantation experiments, the reproduction in vitro or its in vivo characteristics suggest an inherent defect in the nodule; 3) the homogeneity of biochemical findings within each nodule is compatible with the clonality of the lesion; 4) the autonomous nodule is a minimal deviation tumor; and 5) the characteristics of the TSH, cAMP, protein phosphorylation cascade are qualitatively normal, and autonomy does not result from constitutive activation of this system; and 6) a 20K protein, not phosphorylated in response to TSH in the nodule, could represent an absent negative controlling element.

Diversity and prevalence of somatic mutations in the thyrotropin receptor and Gs alpha genes as a cause of toxic thyroid adenomas.

A total of 33 different autonomous hot nodules from 31 patients, originating mainly from Belgium, were investigated for the presence of somatic mutations in the TSH receptor and Gs alpha genes. This constitutes an extension of our previous study, including the first 11 nodules of the series. The complete coding sequence of the TSH receptor gene and the segments of Gs alpha known to harbor mutations impairing guanosinetriphosphotase activity were studied by direct sequencing of genomic DNA extracted from the nodules. DNA from the juxtanodular tissue or peripheral white blood cells was analyzed in all patients to confirm that the mutations identified were somatic. Twenty-seven mutations (82%) were found in the TSH receptor gene, affecting a total of 12 different residues or locations. All these mutations but 2 (see below) have been identified previously as activating mutations. Only 2 mutations were found in Gs alpha (6%). In 4 nodules, no mutation was detected. Five residues (Ser281, Ile486, Ile568, Phe631, and Asp633) were found mutated in 3 or 4 different nodules, making them hot spots for activating mutations. Phe631 and Asp633 belong to a cluster of 5 consecutive residues (629-633) in the N-terminal half of transmembrane segment VI; which harbor together 44% of the mutations identified in this cohort. Two novel mutations were identified: a point mutation causing substitution of Phe for Leu at position 629 (L629F); and a deletion of 12 bases removing residues 658-661 at the C-terminal portion of exoloop 3 (del658-661). When tested by transfection in COS-7 cells, both mutant receptors display increase in constitutive stimulation of basal cAMP accumulation. Although it is still capable of binding TSH, the del658-661 mutant has completely lost the ability to respond to the stimulation by the hormone. Our results demonstrate that, in a cohort of patients from a moderately iodine deficient area, somatic mutations increasing the constitutive activity of the TSH receptor are the major cause of autonomous hot nodules.

Postoperative radiotherapy after pneumonectomy: impact of modern treatment facilities.

PURPOSE: The present study was undertaken to see how modern treatment facilities, computed tomography (CT)-based treatment planning and linear accelerator, have modified the results of postoperative irradiation after a pneumonectomy for lung cancer. METHODS AND MATERIALS: Between 1970-1985, 103 patients were treated in our department after a pneumonectomy: 50 patients with a T1T2N0 tumor and 53 patients with a T3, N1 or N2 tumor. Three groups were considered: 27 patients had only surgical resection, 51 patients were irradiated postoperatively with a Co60 source, and 25 patients were treated using those modern facilities. RESULTS: The 5-year survival varies from 4% to 31% according to the tumor extent but also to the radiation technique. Patients treated with a Co60 source had a dismal 5-year survival rate (8%) whereas patients treated with the modern facilities had a 5-year survival rate of 30% similar to the 31% of the control surgical group including less advanced tumors. CONCLUSION: Linear accelerator and computed tomography-based treatment planning improved the accuracy of postoperative thoracic irradiation and allow to deliver high doses to the mediastinum even after a pneumonectomy.

Ploidy level and proliferative activity measurements in a series of 407 thyroid tumors or other pathologic conditions.

This study describes the ploidy level and proliferation rate in a series of 74 multinodular goiters (MNGs), 17 cases of Hashimoto's disease, 33 cases of Basedow's disease, 113 adenomas, 139 primary carcinomas, and 31 cervical lymph node metastases from 376 patients. Both ploidy level and proliferation rate were assessed by digital cell image analyses of Feulgen-stained nuclei from formalin-fixed, paraffin-embedded tissues. The ploidy level of each sample was assessed using both its DNA index and its DNA histogram type. The proliferation index assessments corresponded to the determination of the proportion of cells in the S-phase fraction. The data reveal that the proportion of aneuploid cases increases according to the following sequence: simple MNGs and normomacrovesicular adenomas-->MNGs with adenomatous hyperplasia and microvesicular adenomas and Hürthle cell adenomas-->papillary and Hürthle cell carcinomas-->follicular and medullary carcinomas-->anaplastic carcinomas. This suggests the preneoplastic nature of the microvesicular adenomas and even of MNGs with adenomatous hyperplasia. The ploidy levels of 99% of the 407 cases of the thyroid tumor series could be described using six DNA histogram types: diploid, hyperdiploid, triploid, hypertriploid, tetraploid, and polymorphic. It was possible to assess the proliferation rate of 279 samples. The results show that a significantly higher proportion of malignant compared with benign thyroid tumors (35.5% v 10.5%, respectively) exhibited a proliferation index higher than 5%, and that, whether benign or malignant, the hypertriploid thyroid tumors proliferated significantly less than the nonhypertriploid thyroid tumors.

Amikacin concentrations in uninfected postthoracotomy pleural fluid and in serum after intravenous and intrapleural injection.

The pharmacokinetics of amikacin after intravenous (IV) and intrapleural injection was compared in 25 patients with pleural drainage after lung resection. In ten patients 7.5 mg/kg of the drug was injected IV; the mean peak concentrations were 31.2 +/- 2.3 micrograms/ml in the serum and 13.3 +/- 3.8 micrograms/ml in the pleural fluid. The penetration of amikacin in the pleural space was 80 percent. After the intrapleural injection of the same dose of amikacin in 15 patients, the pleural fluid concentrations of the drug were extremely high and well sustained during eight hours; however, serum concentrations reached maximal values of 14.1 +/- 4.7 micrograms/ml, indicating a substantial diffusion of amikacin from the pleural space to the blood. In the case of treatment of pleural infections by local injection of aminoglycosides, the serum concentrations must be kept in mind to avoid systemic intoxication from these drugs.

Increased expression of high but not low molecular weight heat shock proteins in resectable lung carcinoma.

Strong expression of high-molecular-weight (HMW) heat-shock proteins (HSP) by lung carcinoma has been documented using immunohistochemistry. Far less is known about the expression of low-molecular-weight (LMW) HSP in lung cancer. We compared the quantitative expression of HMW (HSP-60, HSP-70) and LMW (HSP-27, ubiquitin) HSP in tumor and non-tumor lung tissue obtained from 47 patients undergoing surgical resection of lung carcinoma. HSP levels were determined in cell lysates from tissue samples by ELISA using streptavidin-biotin technology. Results were normalized to total protein content measured by spectrophotometry. Compared to disease-free lung tissue, tumor tissue samples showed higher levels of both HSP-60 (median value: 227 pg versus 96 pg per mg protein (P<0.001 by Wilcoxon Rank test for paired data) and HSP-70 (median value: 525 ng versus 401 ng per mg protein (P=0.01 by Wilcoxon Rank test for paired data). Tumor and tumor-free tissues show similar levels of ubiquitin and HSP-27. Neither the survival rate nor the histologic type and extent of cancer are correlated with the observed differences in HSP-60 and HSP-70 expression (P>0.1 by one way analysis of variance for repeated measures with one between subject factor). Our data confirm, on a quantitative basis, the increased expression of HSP-60 and HSP-70 in non-small-cell lung carcinoma. However, no prognostic value was found to be associated with this over-expression. In contrast, LMW stress proteins such as ubiquitin and HSP-27, although implicated in cellular processes potentially related to malignant transformation, show no increased expression in lung carcinoma.

Induction treatment before surgery for non-small cell lung cancer.

Surgery alone is currently still accepted "standard of care" for patients with operable NSCLC, this includes stages IA and IIB, as well as selected early subsets of IIIA disease. In more advanced and inoperable stage III disease, combinations of chemotherapy and radiotherapy remain the standard treatment approach for patients with good performance status. The role of surgery following induction therapy in these advanced stage III patients is at the moment not conclusively defined. More evidence from randomized trials is clearly needed to tailor treatment for the large number of patients that present in these locally advanced stages. Enrollment of patients into ongoing prospective clinical trials should be encouraged, whenever possible, to further define prognostic factors and improve multimodality strategies in this clinical setting.

The influence of L-triiodothyronine, L-thyroxine, estradiol-17beta, the luteinizing-hormone-releasing hormone, the epidermal growth factor and gastrin on cell proliferation in organ cultures of 35 benign and 13 malignant human thyroid tumors.

PURPOSE: To characterize the influence of six factors on human thyroid tissues at the cell-proliferation level. These six factors were the epidermal growth factor (EGF), the luteinizing-hormone-releasing hormone (LHRH), triiodothyronine, thyroxine, estradiol and gastrin. METHODS: Forty-eight human thyroid specimens were obtained from surgical resection and maintained alive for 48 h ex vivo (in vitro) under organotypic culture conditions. These specimens comprised 35 benign cases (17 multinodular goiters and 18 adenomas) and 13 cancers. Cell proliferation in the control and treated conditions (at a 5 nM dose) was assessed by means of the thymidine labeling index, which enables the percentage of cells in the S phase of the cell cycle to be determined in accordance with autoradiographic procedures. RESULTS: The results show that, of the six factors tested here, EGF significantly (P < 0.05 to P < 0.001) increased cell proliferation in the greatest number of cancers as compared to what happened with the remaining five. Each of these six factors significantly increased or decreased proliferative cell activity in some 10%-30% of the cases under study. CONCLUSIONS: Triiodothyronine, thyroxine, LHRH and gastrin may increase or decrease cell proliferation in human thyroid tissues, whether benign or malignant, to the same extent as other hormones and/or growth factors such as thyrotropin, EGF, insulin-like growth factor 1, transforming growth factor beta1 and estradiol the effects of which on thyroid cell proliferation are already well documented in the literature.

Characterization of ligands for galectins, natural galactoside-binding immunoglobulin G subfractions and sarcolectin and also of the expression of calcyclin in thyroid lesions.

The purpose of this study was to characterize ligands for galectins, natural galactoside-binding immunoglobulin G subfractions and sarcolectin and also the expression of calcyclin in various benign and malignant thyroid lesions. The extent of the binding of eight glycochemical probes was quantitatively assessed using computer-assisted microscopy on 76 thyroid lesions including 10 not-otherwise-specified multinodular goiters (S_MNG), 11 multinodular goiters with adenomatous hyperplasia (AH_MNG), 8 normomacrovesicular (NM_ADE) and 12 microvesicular (MIC_ADE) adenomas, and 9 papillary (P_CAR), 10 follicular variants of papillary (FvarP_CAR), 7 follicular (F_CAR) and 9 anaplastic (A_CAR) carcinomas. The 8 histochemical probes included 5 animal lectins (including galectins and sarcolectin), 1 polyclonal antibody (raised against calcyclin) and 2 immunoglobulin G subfractions from human serum with selectivity to alpha- and beta-galactosyl residues. The results show that multinodular goiters with adenomatous hyperplasia exhibited histochemical characteristics intermediate to those of normal multinodular goiters and microvesicular adenomas. Normomacrovesicular adenomas behaved very distinctly from microvesicular ones. Microvesicular adenomas were more closely related to differentiated thyroid carcinomas than any other type of benign thyroid lesions of epithelial origin. Papillary and follicular carcinomas seemed to represent the two extremes of the same biological entity with the follicular variant of the papillary carcinoma serving as a biological link between these two extremes. Anaplastic carcinomas behaved in a significantly different manner when compared to the differentiated forms of thyroid carcinomas. The results suggest that the patterns of expression of the glycoconjugates investigated in the present study may constitute useful tools for characterizing lesions in the human thyroid.

Synergistic necrotizing cellulitis after pneumonectomy.

Synergistic necrotizing cellulitis is a rapidly progressive infection of subcutaneous tissues and muscles. We report a rare case of synergistic necrotizing cellulitis of the chest wall occurring after a pneumonectomy in a patient without any predisposing factors. Despite rapid and aggressive treatment, the patient died in acute respiratory failure 28 h after the first signs of sepsis.

Randomized trial of surgery versus radiotherapy in patients with stage IIIA (N2) non small-cell lung cancer after a response to induction chemotherapy. EORTC 08941.

Combined modality treatment of patients with stage III non small-cell lung cancer (NSCLC) has recently become widely accepted. Standard combinations are neoadjuvant chemotherapy followed by radiotherapy or concurrent chemotherapy and radiotherapy. The effect of combined modality treatment on survival is dependent on both the efficacy of chemotherapy to eradicate micrometastases and optimal local control. The European Organization for Research and Treatment of Cancer (EORTC) Lung Cancer Cooperative Group has chosen to investigate in a comparative way the side effects and the effect on survival of radiotherapy versus surgery in stage IIIA (N2) NSCLC.

Resection of lung metastases from sarcomas.

Between 1977 and 1987, 19 patients were candidates for resection of lung metastases from pretreated extrathoracic primary tumours. Primary tumours comprised 10 osteosarcomas, one Ewing sarcoma and eight soft tissue sarcomas. All 19 patients presented with metachronous metastases. Twenty-eight thoracotomies were performed in these 19 patients. Nine patients underwent multiple surgical explorations. All the metastases were removed by wedge resection. Seven out of 10 patients treated for osteosarcoma received pre- and postoperative chemotherapy, and three out of 10 postoperative chemotherapy only. The projected survival rate at 3 years is 33%. Seven out of 19 patients survived more than 2 years; four of them were free of disease at 33, 54, 56 and 137 months. Good prognosis appears to be long metastases-doubling time, metachronous metastases, small number of lung metastases, pathological evidence of tumour necrosis and/or fibrosis after chemotherapy and, of course, complete control of the primary tumour and no extrathoracic metastases.

A pilot study for identifying at risk thyroid lesions by means of a decision tree run on clinicocytological variables.

Fine-needle aspiration biopsy (FNAB) is safe, inexpensive, minimally invasive, and highly accurate in the diagnosis of nodular diseases of the thyroid. However, FNAB does not provide a reliable benign versus malignant diagnosis for 100% of the cases analysed. It is possible to increase the accuracy of the cytological diagnosis by means of information contributed by different clinical variables. In the present study we evaluate the diagnostic value of 10 variables in addition to FNAB on a series of 218 specimens for which we obtained histological diagnoses including 37 cancers (17%). The diagnostic information contributed by these variables was analyzed by means of the Decision Tree technique, an artificial intelligence-related method which forms part of the Supervised Learning algorithms. The results show that Decision Trees enable some subpopulations of patients with uncertain FNAB results to be characterized.

[NMR evaluation of neoplastic invasion of the pulmonary artery]. / NMR-Beurteilung neoplastischer Pulmonalarterien-Invasionen.

Bronchial tumors that invade the mediastinum are not necessarily inoperable. Whether surgery is possible depends, among other things, on the extent of pulmonary artery invasion. The authors have studied the value of cardiac-gated MRI and compared it with CT and venous DSA for staging tumor invasion. CT demonstrated the areas of contact between tumor and mediastinum. The MRI planes were transverse and also in the main axis of the pulmonary arteries. Twenty-one patients were studied and in 16 the findings could be checked during surgery. In all cases the findings on MRI were confirmed. In eight patients MRI provided more information than CT and DSA combined and thereby showed its superiority for evaluating arterial invasion.

[Cardio-respiratory assistance with the extracorporeal membrane oxygenator for massive pulmonary embolism]. / Assistance cardio-respiratoire par oxygenateur a membrane extracorporel pour embolie pulmonaire massive

A 39 year old pneumectomized patient presents a massive pulmonary embolism, dies within 3 hours and is supported inefficiently by cardiac massage with recurrent mydriasis during 2 hours. At that time, under extracorporeal cardiopulmonary bypass with a membrane oxygenator, the cardiac activity recovers immediatly due to right decompression and coronary perfusion. The patient is conscious within 5 hours. The cardiopulmonary bypass with a membrane oxygenator appears to be the best therapy when the cardiac massage fails to restitute a normal myocardial function. No embolectomy was performed. The patient died when the bypass was stopped after 48 hours. We conclude that the prolonged peripheral extracorporeal bypass followed by embolectomy is the best therapy of pulmonary embolism.

[Limitations and perspectives of postoperative radiotherapy in bronchial cancer]. / Limites et perspectives de la radiothérapie postopératoire dans le cancer bronchique.

The role of postoperative irradiation for lung cancer remains a controversial issue. The available data suggest a reduction in local relapse in cases of positive mediastinal lymph node, but how this benefit translates into survival is not known. The current indications include tumors with positive mediastinal lymph node and incomplete resection with micro- or macroscopical residue. Nevertheless, postoperative irradiation requires a meticulous technique to avoid inducing life-threatening complications to vital organs such as the heart or the lung.