Mechanism for dexamethasone inhibition of neutrophil migration upon exposure to lipopolysaccharide in vitro: role of neutrophil interleukin-8 release.

The mechanism by which dexamethasone (DEX) inhibits neutrophil (PMN) recruitment to a site of inflammation, such as the newborn lung with bronchopulmonary dysplasia, is not completely understood. The aim of our study was to determine whether DEX inhibits neutrophil-induced neutrophil recruitment by inhibition of interleukin- (IL) 8 release from PMNs, and if there are developmental differences. PMNs isolated from cord blood (CB) and adults (A) were studied. We first measured the effect of DEX (10(-10) to 10(-4) M) on PMN migration to an exogenous IL-8 standard (10(-8) M) using PMNs of CB (n = 3) and A (n = 3), over 1 h in a chemotaxis chamber. Second, we determined the effect of DEX (0 and 10(-10) to 10(-6) M) on IL-8 release (immunoassay) from PMNs of CB (n = 7) or A (n = 7) after incubation with lipopolysaccharide (LPS, 1 ng/mL) for 6 and 18 h. Third, the chemoattractant activity of culture media from the second experiment was studied with and without IL-8 antibody. DEX at concentrations of 10(-10) to 10(-4) M had no direct effect on PMN migration in vitro to an exogenous IL-8 standard. After LPS exposure, IL-8 release was greatly increased for PMNs from CB compared with A. DEX (10(-10) to 10(-4) M) resulted in a dose-dependent inhibition of IL-8 release from PMNs exposed to LPS for 6 and 18 h incubation. Increased PMN migration activity was only found with media of PMNs of CB with no DEX. At 18 h, media-induced migration activity was decreased if DEX (10(-7) M), IL-8 antibody, or DEX (10(-7) M) with IL-8 antibody were present during the incubation with LPS: there was an 88, 86, and 101% reduction in migration activity, respectively. We conclude that DEX inhibits PMN-induced PMN migration, predominantly via inhibition of IL-8 release for PMNs of the newborn. We suggest that a 10-fold lowering of the standard DEX dose may effectively reduce lung inflammation in bronchopulmonary dysplasia.

Elevated urinary leukotriene E4 in chronic lung disease of extreme prematurity.

We determined if pulmonary peptidoleukotrienes contribute to the pathogenesis of chronic lung disease of extreme prematurity (CLD) by measuring urinary leukotriene E4 (uLTE4). Study patients had a birth weight < 1000 g and were about 28 d old when they were classified as normal control subjects (n = 8) or as having CLD (n = 26, abnormal chest X-ray, supplemental O2 requirement +/- ventilator). Urinary LTE4 levels were significantly elevated in CLD compared with the control group (288 +/- 92 versus 35 +/- 10 pg/mg creatinine, mean +/- SE, p < 0.05). Ventilator-dependent CLD patients, who required dexamethasone and had demonstrated uLTE4 levels above the normal range, needed significantly higher peak inspiratory pressures (20 +/- 1 cm H2O versus 15 +/- 1 cm H2O) than similar patients with uLTE4 in the normal range, and the former group had a significant reduction in uLTE4 in the first 5 d of dexamethasone therapy (626 +/- 198 to 451 +/- 176 pg/mg Cr) as ventilatory support was reduced. We conclude that peptidoleukotriene production is activated in patients with CLD (and no other detectable organ dysfunction) to pathophysiologic levels described in adults with acute asthma. Prospective studies focused on infants dependent on high levels of ventilatory support may provide insights into the role of leukotriene synthesis inhibitors or receptor antagonists in the treatment of CLD.

Prolonged pulmonary hypertension caused by platelet-activating factor and leukotriene C4 in the rat lung.

Platelet-activating factor (PAF) and leukotrienes (LTs) are potent pulmonary hypertensive and inflammatory mediators produced by the lung. Previously we showed that a rapid injection of PAF into the pulmonary artery of an isolated rat lung produced an extended elevation in mean pulmonary arterial pressure (PAP). The objective of the present study was to determine whether the extended pressor response induced by PAF was caused by prolonged activation of the 5-lipoxygenase pathway or slow clearance of LTs from the lung parenchyma. Rat lungs were perfused with a nonrecirculating physiological salt solution that contained indomethacin and albumin. Five minutes after a rapid injection of PAF into the pulmonary artery catheter, the following elevations (mean % above baseline) were observed: PAP (83%), LTB4 (3,260%), LTC4 (1,490%), LTD4 (970%), and LTE4 (1,500%). At 20 min these levels declined but were still significantly elevated above baseline. The 5-lipoxygenase inhibitor diethylcarbamazine (DEC), administered before the PAF injection, inhibited the elevations of PAP and all LTs. DEC administration that began 5 min after PAF reduced PAP and only LTC4 levels at 20 min in comparison to lungs with no DEC. The 5-lipoxygenase-activating protein inhibitor MK886, administered orally 2-6 h before perfusion, also inhibited the pressor response to PAF as well as LT production, as did DEC. We conclude that 1) the extended pulmonary hypertension induced by PAF was caused mainly by prolonged activation of 5-lipoxygenase with LTC4 production, 2) the relative overall lung clearance of LTB4, LTD4, and LTE4 was slower than that of LTC4, and 3) LTB4, LTD4, and LTE4 had no appreciable pressor effect.

The effects of endocrine therapy on plasma steroids in prostatic carcinoma patients.

Plasma testosterone (T), 5 alpha-dihydrotestosterone (DHT), T/DHT ratio, estrone (E1), estradiol (E2), E2/T ratio and 17-hydroxyprogesterone (17-OH-P) were measured in 70 patients with prostatic carcinoma (PCA) (T34, N01, M01), after endocrine therapy including: steroidal estrogens (polyestradiol phosphate = Estradurin monthly in injections of 80 mg; estradiol - 17 beta: 5 mg/d), nonsteroidal synthetic estrogens (DES: 5 mg/d; chlorotrianisene = TACE: 24 mg/d; DES-diphosphate = Honvan: 360 mg/d), orchidectomy and their combinations. All these forms of treatment lowered T, DHT, T/DHT ratio and 17-OH-P, the maximal suppression being observed after castration. No further decrease of the mentioned parameters was achieved by estrogen treatment of castrated patients, as compared with castration alone. In the noncastrated patients, steroidal estrogens were less effective than nonsteroidal ones, in terms of lowering T, DHT and T/DHT ratio. Significant decreases in estrone (p less than 0.05) and estradiol (p less than 0.01) levels were observed after nonsteroidal estrogen treatment and castration respectively, and extremely high values of circulating estrogens were found after steroidal estrogen administration.

Estradiol and progesterone receptor levels in human breast cancer in relation to cytosol and plasma estrogen level.

Estradiol receptor (ER) and progesterone receptor (PR) content along with the cytosol and plasma estrone and estradiol levels in 15 premenopausal and 26 postmenopausal women with breast cancer in different clinical stages (T123, N01, M0) were measured. ER-positive tumor frequency and the ER content tended to be higher in postmenopausal than in premenopausal patients. There was no evidence for a relationship between high cytosol estrogen levels and low receptor measurements. The estrogen concentration was higher in ER-positive tumor cytosols than in those of ER-negative tumors; the differences were significant in postmenopausal women, only, with P less than 0.05 for estrone and P less than 0.01 for estradiol values. Twelve pairs of tumor and normal tissue from the same breast, were also studied: seven of which contained ER-positive and five ER-negative tumors. The ER-positive tumors showed a clear trend to higher estradiol content as compared to the corresponding normal tissues. The circulating level of estradiol in postmenopausal women, was higher (P less than 0.05) in ER-positive tumors than that in ER-negative tumors. Our results indicate that: (a) false negative ER assays are not likely to be due to the presence of endogenous estrogens. (b) higher amounts of estrone and estradiol are contained in ER-positive tumors than in negative ones.

Age-related changes of plasma steroids in normal adult males.

Plasma cortisol, 17-hydroxyprogesterone (17-OH-P), testosterone (T), 5 alpha-dihydrotestosterone (DHT, estrone (E1) and estradiol (E2), were measured in 94 normal adult men aged between 20-99, using RIA methods after chromatographic separation of steroids on Sephadex LH-20 columns. All plasma steroids except 17-OH-P, were age dependent: cortisol, testosterone and DHT decreased significantly with age, whereas estrone and estradiol were significantly increased in elderly men. Cortisol, testosterone, T/DHT ratio and estradiol levels were significantly correlated with age. The age related changes of plasma steroids in elderly men, were suggestive of decreased cortisol secretion, and decreased testicular function with increased peripheral conversion of androgens into estrogens. Testosterone was positively correlated with its precursor (17-OH-P) and respectively its peripheral metabolites (DHT and E2). The negative correlation between estrone and 17-OH-P found in elderly men, suggested that increased estrogen level in aging males may be considered able to inhibit the testicular androgen production.

Plasma steroids in benign prostatic hypertrophy and carcinoma of the prostate.

Cortisol, 17-hydroxyprogesterone (17-OH-P), progesterone, testosterone, 5 alpha-dihydrotestosterone (DHT), estrone (E1) and estradiol (E2) were determined by RIA after chromatographic separation of steroids on Sephadex LH-20 columns, in 54 hospitalized patients with benign prostatic hyperplasia (BPH) and in 32 hospitalized patients with prostatic carcinoma (PCA) (T34, N01, M01). The patients' values were compared with those of 63 age-matched controls. Increased cortisol and DHT levels, subnormal estrogen and 17-OH-P values and normal progesterone level were found in both benign and malignant groups. Higher mean values for testosterone, and T/DHT ratio and lower mean values for E2/T ratio were found in PCA, as compared with those in BPH. An age invariance of cortisol, testosterone, T/DHT ratio and estradiol was found in both BPH and PCA, instead of the age dependence found in normal subjects. The normal relations between testosterone and its precursor (17-OH-P) and its peripheral metabolite (E2), respectively, and the normal relation between estrone and 17-OH-P were not evident in either BPH or PCA group. The normal direct relation between testosterone and DHT has been found in both patient groups.

[Importance of hormone dependence of malignant tumors of the breast in orientation of the surgical treatment]. / Importanta hormonodependentei tumorilor maligne mamare în orientarea tratamentului chirurgical.

The authors have investigated the presence of oestrogenic and progesteronic receptors in 38 patients with breast cancer. It was noted that these receptors were present in 20 patients, and that the hormone-dependent tumours are more frequent in patients which are in the post-menopausal stage. The level of circulating oestrogens was also investigated, as well as the amount of oestrogens in the cytosol of tumor cells, and in the normal cells from the diseased breast. In patients with oestrogen-positive tumors, and/or progesteron-positive tumors, a rise was noted in the amount of circulating oestradiol, an increased contents of cytosolic oestradiol, especially in the tumoral cells as compared with the normal ones in the same breast. On the basis of these data patients were selected, with hormone-dependent breast tumors in view of ovarectomy. The indications are discussed, as well as the advantages of surgical ovarectomy.

Adrenal function in early and metastatic breast cancer: dexamethasone suppression of plasma cortisol.

Plasma cortisol, 17-hydroxyprogesterone (17-OH-P), progesterone, FSH, LH and prolactin were determined by RIA, in 14 cancer patients without metastases aged between 40 and 74 years (6 cases of breast cancer: T123, N01, M0 and 8 with other forms of cancer). The cancer patients were investigated: (A) under basal conditions, (B) after three days' adrenal suppression by dexamethasone, 3 mg/day and (C) immediately after local radiation therapy (4500 rads). The basal mean hormonal values in these patients showed increased cortisol, decrease 17-OH-P and normal values of progesterone, FSH, LH and prolactin. Plasma cortisol was significantly reduced by adrenal suppression with a percentage reduction of 19.44 without differences between breast cancer patients and patients with other forms of cancer; adrenal suppression induced an increase of 17-OH-P only in male cancer patients. The only significant hormonal changes after local radiation therapy were increased plasma 17-OH-P values in both female and male cancer patients. A second group investigated in the present study consisted of 11 patients (6 castrated and 5 postmenopausal women) with metastatic breast cancer and presented increased plasma cortisol values, decreased 17-OH-P values, and a great scatter in the estrone values, some of them being very high.

Plasma steroid dynamics in Cushing's syndrome.

The dynamics of five plasma steroids were studied by CPB and RIA in twenty patients with Cushing's syndrome of differing etiologies: pituitary induced adrenal hyperplasia (10), recurrence after adrenal surgery or pituitary irradiation (4), adrenal carcinoma (4) and adrenal adenoma (2). Basal mean values of plasma steroids, showed increase of cortisol in hyperplasia and cancer groups, increase of 17-hydroxyprogesterone (17-OH-P) in recurrence and cancer groups and decrease of testosterone in male patients. In hyperplasia, the changes in 11-desoxycortisol and 17-OH-P (respectively by CPB assay and RIA after LH-20-Sephadex separation) were the reverse of those for cortisol 2 h and 24 h after metyrapone administration. Adrenalectomy determined in all but one patients, significant decrease of cortisol and in male patients, parallel gradual increase of testosterone.

Urinary, plasma and adrenal tissue steroids in a male patient with Cushing's syndrome due to adrenal carcinoma.

The dynamics of urinary and plasma steroids (CPB and RIA) were simultaneously studied in a 36-yr-man with Cushing's syndrome due to adrenal carcinoma. The markedly increased basal excretion of 5-pregnenetriol, PG, PGT, DHA, THS and corticosteroids with nonsignificant changes during the endocrine tests decreased to normal levels after tumour removal. The basal low level of plasma testosterone, with nonsignificant changes during the endocrine tests, rose progressively after surgery. The steroid content of adrenal carcinoma tissue determined by CPB and RIA after LH-20-Sephadex CC separation revealed important proportion of unconjugated 17-OH-P.