Histological changes induced by tibolone and estrogen/glucocorticoid on aging skin.

OBJECTIVE: The aim of this study was to investigate the oral effects of tibolone and mestranol plus paramethasone on the skin of postmenopausal women. A second purpose was to determine endometrial thickness with transvaginal ultrasound. MATERIALS AND METHODS: This randomized study was carried out in 39 healthy postmenopausal women. Skin biopsies were obtained from the thigh area by a single punch, before and after treatment, and the sections were evaluated. Current characteristics of both groups were measured at follow-up. RESULTS: No gross differences were observed in size, distribution or imaging of collagen, elastic or reticular fibers. Statistically significant changes were found in the papillar dermis thickness. There were no statistically significant differences in the sonographic measurements. CONCLUSION: The estrogen/glucocorticoid combination provides a way to evaluate in parallel the cellular metabolism effects on the irreversible aging process. The current results encourage widening these observations of the possible advantage of this combination, in order to alleviate the cellular degenerative process.

Inverted skin changes induced by estrogen and estrogen/glucocorticoid on aging dermis.

To evaluate hormone replacement therapy effects at the peripheral level, the present trial explored the oral effects of mestranol (80 micrograms/day for 10 days) and mestranol plus paramethasone (80 micrograms + 6 mg/day for 10 days) on the skin of postmenopausal women. This double-blind study included 13 patients. Skin biopsies were obtained from the thigh area by a single punch, 5 mm in diameter, before and after treatment, and the sections, six per sample, were micrometrically evaluated. Various features of the epidermis and dermis layers were stained using the hematoxylin and eosin, Masson, Verhoeff's and Gomori techniques. Statistically significant changes were found in the papillar dermis thickness; mestranol reduced it and mestranol plus paramethasone increased it. The current results encourage widening these observations of the possible advantage of this estrogen/glucocorticoid combination, in order to alleviate the cellular degenerative process.

Paramethasone acetate (PA): corticosteroid potency vs hypothalamic pituitary-gonadal axis.

Because paramethasone acetate (PA) suppresses basal and midcycle LH surge and blocks estrogen synthesis in the female, its possible effect upon testicular physiology was evaluated in 13 healthy men by measuring the circulating levels of FSH, LH, prolactin (PRL), testosterone (T), dihydrotestosterone (DHT), androstenedione (A), estradiol (E2) and cortisol (C) every 4 h throughout the day, before (control) and after PA (6 mg/d/7 d). The total concentrations of each hormone, as well as the PA-induced suppressibility (measured as percent decrease in the mean 24 h plasma level) were analyzed. PA suppressed neither the basal nor circadian rhythm of T and had no effect on LH, FSH or PRL output. DHT, A, E2 were significantly reduced and the basal concentrations and circadian variations of C were abolished. PA showed a dual control on the pituitary gonadal axis and while causing a maximal suppressed adrenocortical activity it had no interference in testosterone synthesis.

Effect of paramethasone acetate upon estrogen action.

To test the antiestrogenic action of paramethasone acetate (PA), a group of five postmenopausal women were treated in two ways. For phase I, PA + mestranol, 6 mg + 80 micrograms per day for 10 days, was administered. Ten days were allowed for withdrawal of medication. For phase II, mestranol, 80 micrograms per day for 10 days, free of the glucocorticoid was administered. Daily samples of cervical mucus, vaginal cells, and peripheral blood were obtained to analyze fernlike crystallization (%), cornified pyknotic nuclei cells (%), and concentrations of ethynyl-estradiol (EE-2, pg/ml) during both phases. The fernlike crystallization pattern had a mean value of 8 +/- 0.9% for phase I, while for phase II it was 85 +/- 8% (P less than 0.05). The percentage mean value of cornified pyknotic nuclei cells of phase I was 28 +/- 5% in comparison with 53 +/- 7% observed in phase II (P less than 0.05). EE-2 mean value concentrations for phase I were 85 +/- 90 pg/ml, while for phase II they were 724 +/- 418 pg/ml (P less than 0.05). These results demonstrate that 6 mg of PA is able to compete against the estrogenic action in the parameters of the study selected.

Demonstration of immunity against Isospora suis in swine.

Piglets naturally exposed or experimentally infected with Isospora suis oocysts were given challenge doses of oocysts to determine the extent of development of immune resistance. Piglets in both studies shed low numbers of, or no detectable oocysts, following challenge. Administration of methylprednisolone acetate failed to induce oocyst shedding in previously infected piglets. Piglets rechallenged with I. suis following steroid injections also failed to shed significant numbers of oocysts suggesting development of immunity to reinfection.

Release of chondrocyte-stimulating factor by rabbit peritoneal macrophages.

Cultured rabbit peritoneal macrophages, after stimulation with lipopolysaccharides (LPS), produce a factor that induces normal rabbit articular cartilage cells (chondrocytes) to release collagenase and other neutral proteases in their culture medium. The release of the factor as well as the activation of chondrocytes can be significantly inhibited by paramethasone (10(-6) M). Rabbit peripheral blood monocytes produce this factor in smaller quantities. Activation with LPS does not enhance the release of factor any further by these cells. Lymphocytes have no direct effect on the chondrocytic protease synthesis. Furthermore, conditioned medium of activated lymphocytes failed to stimulate monocytes or macrophages in the absence of LPS. The macrophage medium exhibits mitogenic and phytohaemagglutinin-enhancing activity towards thymocytes of C3H/HeJ mice, but not against species-specific rabbit lymphocytes. The lymphocyte-activating factor, derived from a mouse macrophage cell line, P388D1 cells, or from other sources, was unable to stimulate chondrocytic protease secretion. Such specific induction of chondrocytic proteases by a macrophage-derived factor may have an important role in cartilage destruction in arthritic conditions, where synovium is only marginally involved.

Glucocorticosteroid inhibition of nonphagocytic discharge of lysosomal enzymes from human neutrophils.

Glucocorticosteroids and mineralocorticosteroids were tested for their capacity to inhibit the nonphagocytic discharge of two lysosomal enzymes--a cartilage matrix-degrading neutral protease and beta-glucuronidase--from highly purified human neutrophils. Lysosomal enzyme discharge from neutrophils adherent to nonphagocytizable, immobilized, heat-aggregated IgG was inhibited by the four glucocorticosteroids--methylprednisolone sodium succinate, triamcinolone acetonide hemisuccinate, para methasone acetate, and hydrocortisone sodium succinate. These glucocorticoids also inhibited zymosan-induced release of beta-glucuronidase from neutrophils that had been pretreated with cytochalasin B in order to completely prevent the onset of phagocytosis. Inhibition by the glucocorticoids of lysosomal enzyme discharge provoked by a soluble divalent cation ionophore was also observed. Neither desoxycorticosterone acetate nor aldosterone hemisuccinate, two mineralocorticosteroids, inhibited lysosomal enzyme release. Similarly, the salt moieties of some of the steroids tested, such as sodium succinate and sodium acetate, failed to elicit an effect on enzyme release. Therefore interference with lysosomal enzyme discharge was restricted to the glucocorticosteroid ring structure. Because interference either with the adherence of neutrophils to immune reactants or with the activities of the discharged lysosomal enzymes by the glucocorticoids could be interpreted as inhibition of lysosomal enzyme release, steroidal effects on these parameters were examined. None of the glucocorticoids tested elicited any significant effects on neutrophil adherence or lysosomal enzyme activity. Thus it appears that glucocorticosteroids are capable of inhibiting directly the nonphagocytic discharge of lysosomal enzymes from human neutrophils.

Effect of paramethasone acetate on ovarian steroids and gonadotropins. I. Normal menstrual cycle.

In an attempt to elucidate the effect of paramethasone acetate on the hormonal profile of the normal menstrual cycle, 5 clinically healthy women, aged 24-36 yr, with ovulatory cycles were studied during a control menstrual cycle and during treatment with paramethasone acetate (6 mg/day). The length of the untreated cycle was 29.2 +/- 2.3 days as compared to 29.0 +/- 3.1 days during the paramethasone-treated cycle. Peak plasma E-2 level was 188.2 +/- 42.1 pg/ml in the control cycle and 74.4 +/- 15.3 pg/ml during paramethasone treatment (P is less than 0.05). Peak plasma 17 OH-P was 4.0 +/- 0.4 ng/ml in the control cycle and 1.6 +/- 0.2 ng/ml during the paramethasone treated cycle (P is less than 0.005). No significant differences in plasma progesterone were observed during the luteal phase of both cycles. Minimal and scattered differences were observed in plasma FSH. However, plasma LH levels were lower in the paramethasone acetate than in the control cycle almost throughout the entire period of study. Furthermore, midcycle LH peak in the control cycle was 958 +/- 104 ng/ml as compared to 283 +/- 24 ng/ml during the paramethasone treatment (P is less than 0.005). Despite these differences, ovulation occurred during paramethasone treatment based upon the observed rise in BBT, the plasma progesterone levels above 6 ng/ml and the secretory changes in the endometrium. These results suggest that: 1) Paramethasone may block E-2 synthesis at the ovarian level and , 2) Ovulation may still occur even in the presence of E-2 and LH plasma concentrations lower than those occurring in the normal menstrual cycle.

Effects of corticosteroid treatment and inflammation on the cellular content of blood and exudate in mice.

Inflammatory exudates have been produced in mice by intraperitoneal injection of thioglycollate broth 24 hr and again 3 hr before collecting of the exudate. The first injection of broth exerts a "priming" effect leading to an enhanced response to the second injection. By this procedure more than 30 x 106 leucocytes of which 78 per cent. were polymorphs were obtained from each mouse. Administration of paramethasone reduced the number of cells in the exudate when given at the same time as broth but not when given 3 hr before collection of the exudate. In contrast paramethasone was equally effective when given either 24 hr or 3 hr before harvest in suppressing the appearance of intravenously injected pontamine sky blue in the exudate. It was striking that treatment with paramethasone which had reduced the number of polymorphs in the exudate had actually increased the number in blood.

[Effect of high dosis of D-penicillamine and paramethasone on mechanical properties of rat connective tissue (author's transl)]. / Einfluss hoher dosen von D-Penicillamin und Paramethason auf die mechanischen Eigenschaften des Bindegewebes der Ratte

Male albino rats are treated with high doses of D-penicillamine (2000 mg/kg/day orally), paramethasone (50 mu-g/kg/day orally), and with a combination of equal doses of D-penicillamine and paramethasone for a period of 4 weeks. At the end of treatment the mechanical properties of dorsal skin and aorta of the experimental animals are determined. 4 weeks of treatment with D-penicillamine or paramethasone, respectively, leads to a decrease in the skin thickness of the experimental animals. With simultaneous administration of both substances the decrease in skin thickness is about the same. Load resulting in rupture is considerably reduced by D-penicillamine, while only a slight decrease is seen with paramethasone. Simultaneous administration of both substances results in considerable decrease in load resulting in rupture, similar to the effect produced by D-penicillamine. Tensile strength which is not influenced by paramethasone treatment, decreases after treatment with D-penicillamine as well as after simultaneous administration of D-penicillamin and paramethasone. Load resulting in rupture of the aorta remains uninfluenced under the experimental conditions. Comparison of the groups shows that simultaneous administration of D-penicillamine and paramethasone leads to the same results as those which may be obtained with sole administration of D-penicillamine. This suggests that the effect of D-penicillamine on the mechanical properties of the rat skin is not influenced by an additional high dose of paramethasone under the experimental conditions chosen.

Lysosomal enzyme secretion from human neutrophils mediated by cyclic CMP: inhibition of cyclic GMP accumulation and neutrophil function by glucocorticosteroids.

The effects of several glucocorticosteroids on cyclic GMP accumulation, guanylate cyclase activity, calcium influx, lysosomal enzyme secretion, and phagocytosis were studied in human neutrophils. Contact between neutrophils and serum-treated zymosan particles, in the presence of calcium at pH 7.4, triggered these cellular events within five minutes. Each of these neutrophil functions was markedly inhibited by methylprednisolone sodium succinate, triamcinolone acetonide hemisuccinate and paramethasone acetate but was unaffected by two mineralo-corticosteroids. Human neutrophil soluble guanylate cyclase activity was not changed by the glucocorticoids. Inhibition of phagocytosis by, and lysosomal enzyme secretion from, neutrophils by glucocorticosteroids may be the result of a reduction in cyclic GMP accumulation within these cells. The data suggest that glucocorticosteroids inhibit cyclic GMP accumulation in neutrophils by reducing the influx of extracellular calcium into the cells, thereby limiting the availability of intracellular calcium for metabolic processes associated with the accumulation of cyclic GMP.