Induction of intercellular adhesion molecule 1 (ICAM-1) expression in normal human eosinophils by inflammatory cytokines.

Intercellular adhesion molecule-1 (ICAM-1) functions as a ligand for lymphocyte function-associated antigen-1 (LFA-1), and thereby plays a crucial role in mediating cell-cell interactions in inflammatory reactions. Human eosinophils represent important effector cells in allergic skin diseases. To gain more insight into the capacity of eosinophils to physically interact with LFA-1-positive inflammatory leukocytes, in the present study ICAM-1 expression in eosinophils was investigated. Using fluorescence-activated cell sorter analysis, it could be shown that highly purified (> or = 95%) eosinophils from peripheral blood of non-atopic individuals do not constitutively express ICAM-1 molecules. However, stimulation of eosinophils with interferon gamma (IFN gamma), tumor-necrosis factor alpha (TNF alpha), or interleukin 3 (IL-3) markedly upregulated ICAM-1 surface expression in a time- and dose-dependent manner. Cytokine-induced ICAM-1 expression in human eosinophils was corroborated by Northern blot analysis. Accordingly, unstimulated eosinophils did not express significant amounts of ICAM-1 mRNA, but ICAM-1 mRNA expression could be markedly induced in these cells upon stimulation with IFN gamma plus TNF alpha. The combination of TNF alpha with either IFN gamma, IL-3, IL-5, or granulocyte/macrophage colony-stimulating factor (GM-CSF) increased ICAM-1 expression in a synergistic fashion, whereas IL-5 or GM-CSF by itself did not induce ICAM-1 expression. Cytokine-induced ICAM-1 expression was specific, because IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-7, IL-8, C5a, and platelet-activating factor did not significantly affect eosinophil ICAM-1 surface expression. In summary, these studies indicate that eosinophils may be activated to express the adhesion molecule ICAM-1 upon stimulation with selected inflammatory cytokines, which may allow adhesion-mediated cross-talk between eosinophils and LFA-1-positive cells. In addition, these data demonstrate for the first time a role for IL-3, IL-5, and GM-CSF in regulation of ICAM-1 expression in human cells.

Evidence of the involvement of phosphatidylinositol 3-kinase in the migration, actin stress fiber formation, and alpha v beta 3-integrin-mediated adherence of human melanoma cells.

Tumor invasion and formation of metastases are major obstacles for a successful therapy of melanomas. Metastasis is thought to require multiple steps such as alpha v beta 3-integrin-mediated adhesion, proteolytic digestion of extracellular matrix by metalloproteinase-2, and reorganization of the actin cytoskeleton. To analyze the functional role of phosphatidylinositol 3-kinase in these processes, melanoma cells were treated with the fungal metabolite wortmannin. Wortmannin inhibited phosphatidylinositol 3-kinase activity in melanoma cells and migration in an equally concentration-dependent fashion. Flow cytometric analysis of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phallacidin-stained actin network indicated reduction of actin filaments by wortmannin. Fluorescence laser confocal microscopy experiments revealed breakdown of actin stress fibers. In addition, wortmannin inhibited alpha v beta 3-integrin-mediated adhesion of melanoma cells to vitronectin. Since flow cytometric measurements did not show altered expression of the alpha v beta 3-integrin at the cell surface, avidity changes of the alpha v beta 3-integrin by wortmannin are suggested. In contrast to the actin analysis and adhesion assays, wortmannin had no influence on mRNA expression or on protein secretion of metalloproteinase-2. These data provide evidence that phosphatidylinositol 3-kinase is an essential signal transduction protein required for migration of melanoma cells, regulating formation of the actin stress fiber as well as alpha v beta 3-integrin-mediated adhesion.

The chemokine RANTES is more than a chemoattractant: characterization of its effect on human eosinophil oxidative metabolism and morphology in comparison with IL-5 and GM-CSF.

Eosinophils were shown to play a major role in the allergic inflammatory process leading to the clinical symptoms of atopic dermatitis. Only selected cytokines are capable of inducing a chemotactic response in eosinophils. In particular, the chemokine RANTES was recently shown to be a potent eosinophil chemotaxin. To examine the role of RANTES in eosinophil activation, we investigated the effect of RANTES and other chemokines on morphology and oxidative metabolism of highly purified eosinophils of normal nonatopic blood donors by assessment of functional as well as morphologic criteria. RANTES, and, to a lesser extent, MIP-1 alpha significantly induced the production of reactive oxygen species by human eosinophils, whereas MCP-1, MIP-1 beta, and interleukin (IL)-8/NAP-1 had no significant effects. RANTES stimulated only a subpopulation of the normal eosinophils. With the exception of IL-8, none of the cytokines tested had any significant effect on polymorphonuclear neutrophilic granulocytes. By scanning electron microscopy, RANTES induced characteristic changes that were completely abrogated in the presence of cytochalasin B. Based on functional and ultrastructural assays significant extracellular but not intracellular H2O2 production was detected and completely inhibited by cytochalasin B. Separation of eosinophils by discontinuous density gradients revealed the existence of two hypodense eosinophil populations, one which showed significantly reduced responses upon stimulation with RANTES. RANTES-induced production of reactive oxygen species was almost completely inhibited by staurosporine, wortmannin, or pertussis toxin. Based on these data it is evident that RANTES represents a potent eosinophil-specific activator of oxidative metabolism. Besides its chemotactic activity on T cells and eosinophils, therefore, RANTES may be involved in the functional activation of eosinophils in the skin of patients with atopic dermatitis.

Ultraviolet radiation effects on human keratinocyte ICAM-1 expression: UV-induced inhibition of cytokine-induced ICAM-1 mRNA expression is transient, differentially restored for IFN gamma versus TNF alpha, and followed by ICAM-1 induction via a TNF alpha-like pathway.

Human keratinocytes (KC) during the course of inflammatory dermatoses strongly express the surface molecule ICAM-1, which plays an important role in the generation of the epidermal inflammatory infiltrate by mediating leukocyte-keratinocyte interactions. Accordingly, KC ICAM-1 expression is known to be induced in vivo and in vitro by cytokines either via the TNF alpha/TNF beta or via the IFN gamma-mediated pathway. In contrast, ultraviolet (UV) radiation previously has been found to potently inhibit cytokine-induced KC ICAM-1 surface expression by a sublethal mechanism. In order to further define this novel immunosuppressive effect of UV light, the effects of in vitro UV radiation on ICAM-1 mRNA expression in transformed human KC (KB cells) were examined. Accordingly, UV light (0-100 J/m2) inhibited IFN gamma- as well as TNF alpha-induced ICAM-1 mRNA expression, if KC were cytokine stimulated immediately after irradiation. After a 12-h incubation period, however, IFN gamma responsiveness was found to be restored in irradiated cells, whereas restoration of responsiveness to TNF alpha required at least a 24-h recovery phase. Moreover, UV light alone did not alter ICAM-1 mRNA levels after 4, 12, or 24 h. After 48 h, however, a significant increase in ICAM-1 mRNA and surface expression in UV-irradiated KC could be observed. In addition, this increase could be superinduced by stimulation of irradiated KC with IFN gamma, but not with TNF alpha. UV-induced upregulation of ICAM-1 expression could be mimicked by stimulating unirradiated cells with supernatants derived from UV-irradiated cells. Addition of biologically active anti-TNF alpha antibodies to UV-irradiated cells or to supernatants derived from UV-irradiated KC, however, did not even partially abolish this ICAM-1-inducing activity. UV light thus seems to affect KC ICAM-1 mRNA expression in a biphasic manner: an early period of inhibition of cytokine-induced ICAM-1 expression is transient and followed by restoration of responsiveness to ICAM-1-inducing cytokines. Moreover, UV itself is able to induce ICAM-1 mRNA expression at this later time point via a TNF alpha-like pathway. These studies identify UV irradiation as a potent modulator of cytokine regulated ICAM-1 gene transcription with the capacity to induce both inhibitory as well as enhancing effects.

Analysis of the cytokine pattern expressed in situ in inhalant allergen patch test reactions of atopic dermatitis patients.

In the present study, we examined the cytokine pattern expressed in situ during the development of eczematous reactions that had been provoked in atopic dermatitis patients by patch testing with house dust mite allergen. In 24-h house dust mite allergen patch test reactions, expression of interleukin (IL)-4 mRNA and IL-2 mRNA increased, but interferon (IFN)-gamma mRNA did not, as compared with control skin. In 48-h inhalant allergen patch test reactions, however, IFN-gamma mRNA and IL-2 mRNA expression were increased above levels observed in control skin, whereas IL-4 mRNA expression was decreased below background levels. These data demonstrate that a switch from a Th2-like to a Th1-like cytokine response occurs in inhalant allergen patch tests of atopic dermatitis patients. This biphasic pattern was specific to inhalant allergen patch test reactions, as it was not observed in irritant reactions in the same patient. IFN-gamma production by T cells may be induced by the cytokine IL-12. In the present study, up-regulation of IFN-gamma mRNA expression in inhalant allergen patch test reactions was preceded by an increased expression of the p35 subunit of IL-12 mRNA. These observations suggest that increased IL-12 expression may contribute to the observed switch of the in situ cytokine secretion pattern. Further studies are necessary to determine whether a similar biphasic pattern of cytokine expression is also operative in the initiation phase of atopic eczema.

Chemotactic 5-oxo-eicosatetraenoic acids induce oxygen radical production, Ca2+-mobilization, and actin reorganization in human eosinophils via a pertussis toxin-sensitive G-protein.

The arachidonic acid metabolites 5-oxo-[6E,8Z,11Z,14Z]-eicosatetraen oic acid (5oETE) and 5-oxo-15-hydroxy-[6E,8Z,11Z,13E]-eicosatetrae noi c acid (5oHETE) are potent eosinophil chemotaxins. Here, the activation profile of 5-oxo-eicosanoids in eosinophils was further characterized and compared to other eosinophil activators such as complement fragment C5a (C5a), platelet-activating factor (PAF), interleukin-5 (IL-5), and phorbol ester (PMA). Flow cytometric studies revealed a rapid and transient actin polymerization upon stimulation by both 5-oxo-eicosanoids. Desensitization studies using actin polymerization as the parameter indicated cross-desensitization between the two 5-oxo-eicosanoids but revealed no interference with the response to other chemotaxins. Fluorescence measurements with Fura-2-labeled eosinophils in the presence of EGTA indicated Ca2+-mobilization from intracellular stores by 5oETE and 5oHETE. Both 5-oxo-eicosanoids stimulated the production of reactive oxygen metabolites as demonstrated by lucigenin-dependent chemiluminescence, superoxide dismutase-inhibitable cytochrome C reduction, and flow cytometric dihydrorhodamine-123 analysis. At optimal concentrations the changes induced by 5-oxo-eicosanoids were comparable to those obtained by C5a and PAF, whereas IL-5 and PMA induced only a restricted pattern of cell responses. Cell responses elicited by 5-oxo-eicosanoids were inhibited by pertussis toxin, indicating coupling of the putative 5-oxo-eicosanoid-receptor to G-proteins. These results indicate that 5-oxo-eicosanoids are stong activators of eosinophils with comparable biologic activity to the eosinophil chemotaxins C5a and PAF. These findings point to a role of 5-oxo-eicosanoids in the pathogenesis of eosinophilic inflammation as chemotaxins as well as activators of pro-inflammatory activities.

High prevalence of thyroid autoimmunity in idiopathic Addison's disease.

We have determined the incidence of autoimmune thyroid disorders in patients with Addison's disease. The material comprised 212 patients, 128 women and 84 men, aged 9-74 years. In 58 patients tuberculosis and in six patients other adrenal disorders were diagnosed. In the remaining 148 patients the auto-immune mechanism was the most probable cause of adrenocrotical insufficiency. In order to evaluate the thyroid abnormalities seen in patients with Addison's disease the T3, T4 (RIA), TSH (ELISA) and anti-thyroid autoantibodies were determined apart from routine clinical examination. Antimicrosomal, anti-thyroglobulin and anti-thyroperoxidase antibodies were measured in 91, 188 and 81 cases respectively. Thyrotoxicosis was diagnosed in 17 and primary hypothyroidism in 18 cases. Moreover, eight patients had evidence of subclinical hypothyroidism. The anti-thyroglobulin antibodies with titer ranging from 1:80 to 1:10,000 were detected in 66 patients, whereas antimicrosomal antibodies were found in 51 patients at a titer ranging from 1:80 to 1:9720. Autoantibodies against thyroid peroxidase were found in sera of 67 patients, with titer ranging from 1:2000 to 1:25,6000.

[The radiological and scintigraphic findings in psoriatic arthropathy (author's transl)]. / Röntgen- und szintigraphische Befunde bei Psoriasis arthropatica

The radiological and scintigraphic findings in the skeletons of patients with psoriasis and psoriatic arthropathy are described. The results indicate that is it possible to detect inflammatory disease in joints by scintigraphy in patients who have not yet developed symptoms. The effect of treatment can be satisfactorily followed by serial scans. Further investigations and observations will be required to show whether the paravertebral calcification and syndesmophyte formation which has been described is really pathognomonic for psoriatic arthropathy.

[Clinical evaluation of the antioestrogenic agent tamoxifen in the treatment of metastatic breast cancer (author's transl)]. / Klinische Beurteilung der Antiöstrogensubstanz Tamoxifen in der Behandlung des metastasierenden Mammakarzinoms.

The endocrine management of advanced breast cancer is of great clinical importance. Apart from the known therapeutic approach with oestrogens, androgens, gestagens and steroids, the oestrogen antagonist, tamoxifen appears to be a promising therapeutic agent. In this study from October 1st, 1975, to October 1st, 1976, the value of tamoxifen was examined in a controlled clinical trial. An objective remission after 3 months' treatment was achieved in 41% of the cases. The criteria of Karnofsky were used to evaluate clinical success. The median time of remission was 5,5 months. The determination of oestrogen receptors could provide a better selection of patients and, hence, help to achieve an increase in the remission rate. An interesting observation was a probably immunostimulative effect, but further investigations are necessary. The side-effects of treatment are detailed and the indications for the use of tamoxifen are discussed. More astonishing than the objective remission was the subjective improvement with relief of pain due to generalized bone metastases. On the basis of our observations it is concluded, that the use of the oestrogen antagonist, tamoxifen presents a potent alternative in the treatment of advanced and incurable breast cancer, especially in postmenopausal patients.

[Studies in the diagnosis of breast disease (author's transl)]. / Vergleichende Untersuchungen zur Mammadiagnostik

The relative diagnostic accuracy of clinical examination, mammography, thermography and ultrasound was investigated in a comparative study. Mammography proved to be the most accurate diagnostic method, followed by clinical examination, thermography and ultrasound. Whereas in cases of advanced cancer (T2 to T4 tumour diameter greater than 2 cm) the correct diagnosis was made most reliably by clinical examination, mammography was superior to all other procedures in T1 tumours and in impalpable tumours. In our opinion thermography and ultrasound should be excluded for routine use or as a screening test because of the high false negative results. It is, however, noteworthy, that small intraparenchymal cysts can be detected and localized by ultrasound in a very high percentage of cases.

[Modern methods in the diagnosis of breast disease (author's transl)]. / Moderne Methoden der Mammadiagnostik

240 patients attending a surgical clinic for breast diseases were examined clinically and subjected to mammography, thermography and ultrasonic investigation. Histological reports are available in 98 cases, enabling an evaluation of the accuracy of the used diagnostic methods. The most efficient diagnostic method is X-ray mammography, followed by thermography and ultrasound diagnosis, but these procedures are still fraught with methodological and technical problems.

[Clinical aspects, radiology and therapy of angiosarcoma of the breast gland]. / Klinik, Radiologie und Therapie des Angiosarkoms der Brustdrüse.

A case of angiosarcoma of the breast is described. The problems of clinical appearance, roentgenology and therapy are discussed. The case described is similar to those seen in the literature, of which 44 cases are known in the literature. Problems of diagnosis even during surgery are evident.

[Effect of the oral contraceptive preparation Femigen on the erythrocyte and resin uptake of 131-I-triiodothyronine]. / Wplyw preparatu femigen stosowanego dla celów antykoncepcyjnych na wartosci wychwyu erytrocytowego i zywicowego 131-J-trójjodotyroniny.

PIP: 150 women, 50 in the 1st and 3rd trimesters of normal pregnancy, 50 in the 6th month of curation administered by Femigen-forte, and the same 50 women 3 months after discontinuation of the use of the preparation, were given erythrocyte and resin up-take tests for labeled triiodothyronine. The values obtained from the women who had used the Femigen preparation were similar to the values obtained from those women in normal gestation. The results of the group of women who had discontinued the use of Femigen normalized, although there were some age-linked differences within this group; results in women aged 31-42 years were slightly lower than in those women aged 20-30 years.^ieng

[Changes in some metabolic pathways in women ingesting progestagens as contraceptives]. / Zmiany w niektórych metabolizmach ustrojowych kobiet stosujacych preparaty gestagenowe dla celów antykoncepcyjnych

PIP: Literature concerning the changes caused in lipids and protein metabolism in women using gestagen preparations for contraceptive purposes is surveyed. The level of metabolic changes was found to be related to the estrogen content of the preparation. Lipid metabolism changes are in the form of increases in the total lipids in the serum, insignificant increases in the beta-lipoproteins, and increases in the total cholesterol concentrations. The preparations Femigen-Polfa and Anacyclin were not found to increase the total cholesterol concentrations in the pre beta and beta lipoprotein fractions. There is a possible effect on the triglyceride concentrations. Gestagens are assumed to cause changes in the blood serum protein fractions characterized by an increase in the alpha-1, alpha-2, and beta globulins. Concentrational and transportational changes are caused by decrease of the erythrocytes and resin test values and increase in the thyroid hormone binding globulins with a collateral normal concentration of free thyroxine and normal index of free thyroxine. These changes are observed only with the use of estrogen preparations.^ieng

[Pregnancy in women with thyroid cancer treated with suppressive doses of L-thyroxine]. / Ciaza u kobiet z rakiem tarczycy leczonych supresyjna dawka L-tyroksyny.

UNLABELLED: The aim of the study was to estimate the dose of thyroxine required by pregnant women who had undergone total thyreoidectomy and radioiodine treatment for thyroid cancer. Material consisted of 4 pregnant women, aged mean 30.8 years. One of patients was studied during 2 consecutive pregnancies. The daily mean dose of thyroxine was 175 micrograms. The control group consisted of 7 women with primary hypothyroidism aged mean 33.5 years, who were treated with replacement doses of thyroxine. One of them was pregnant twice. The mean daily dose of thyroxine was 106.3 micrograms. The estimation of TSH, fT4 were repeated every 4 weeks. RESULTS: In all cases natural deliveries took place. All infants were alive and had no congenital malformations and no clinical or biochemical thyroid dysfunction was found. Pregnant women treated for thyroid cancer needed to have optimized their suppressive therapy by increasing the dose of thyroxine by 26% at the first trimester, 27% at the second and 38% at the last one. Statistically significant increase was found at the 1st trimester of pregnancy and it remained at the same level till the delivery. Pregnant hypothyroid women needed to have optimized their replacement thyroxine therapy by increasing of the dose by 53% at the first trimester, by 49% at the second and by 53% at the last one. Similarly to the 1st group of patients, we noticed statistically significant increase at the 1st trimester of pregnancy. CONCLUSION: In pregnant women who have been previously treated for thyroid cancer the suppressive dose of thyroxine needs to be increased by 26-38% which is slightly less than the increase of the replacement dose in hypothyroid pregnant women.

[Determining serum TBG level using the charcoal-cellulose I-125 thyroxine uptake test]. / Oznaczanie poziomu tbg w surowicy na podstawie weglowo-celulozowego wychwytu 125I-tyroksyny.

Radiocompetition method for the determination of blood serum TBG level has been presented. The method is based on binding of 125I-thyroxine by the mixture of cellulose and activated charcoal following the saturation of TBG with L-thyroxine at the concentration of 2.9 nmol/l. The method is similar to the previously described labeled thyroxine uptake in many respects including the use of the same reagents and equipment. The method is simple, easily accessible for any radiochemical laboratory, and yields satisfactory results. The values of TBG obtained with the described method cas serve, together with the values of total thyroxine concentration, for the calculation of free thyroxine index (as TT4/TBG). The above index differentiates well various functional states of the thyroid and serves as a good measure of free thyroid hormones in euthyreosis accompanied by variable TBG values.