The role of inflammatory markers in assessing disease severity and response to treatment in patients with psoriasis treated with etanercept.

BACKGROUND: Psoriasis is a chronic, systemic, inflammatory disease. Inflammatory markers are used in clinical practice to detect acute inflammation, and as markers of treatment response. Etanercept blocks tumour necrosis factor (TNF)-α, which plays a central role in the psoriatic inflammation process. AIM: To reveal any possible association between disease severity [measured by Psoriasis Area and Severity Index (PASI)] and the inflammatory burden (measured by a group of inflammatory markers), before and after etanercept treatment. METHODS: In total, 41 patients with psoriasis vulgaris, eligible for biological treatment with etanercept, were enrolled in the study. A set of inflammatory markers was measured, including levels of white blood cells and neutrophils, fibrinogen, ferritin, high-sensitivity C-reactive protein (hs-CRP), erythrocyte sedimentation rate (ESR), haptoglobin, ceruloplasmin and α1-antitrypsin, before and after 12 weeks of etanercept 50 mg twice weekly. RESULTS: All markers were reduced after treatment (P < 0.001). PASI correlated with fibrinogen and hs-CRP. Of the 41 patients, 19 (46.3%) achieved reduction of 75% in PASI (PASI75). An increase in hs-CRP and ESR difference (values before minus values after treatment) was related to higher likelihood of achieving PASI75. CONCLUSIONS: Inflammatory markers, particularly hs-CRP and to a lesser extent, fibrinogen and ESR, can be used to assist in assessing disease severity and response to treatment in patients with psoriasis. A combination of selected inflammatory factors (which we term the Index of Psoriasis Inflammation) in combination with PASI might reflect inflammatory status in psoriasis more accurately than each one separately.

Serum adiponectin and resistin in rats under three isocaloric diets: The effect of sibutramine.

The aim of the study was to investigate the effects of (1) macronutrients on food intake, body composition and serum resistin and adiponectin and (2) sibutramine(S) on the above parameters in rats fed with isocaloric diets. Three groups of male Wistar rats (n=63) were fed with high fat diet (HFD), high carbohydrate diet (HCD) or high protein diet (HPD) for 13weeks. In the last 3weeks each group was divided into three subgroups and received S 5mg/kg or 10mg/kg, or vehicle. Body weight was measured weekly, gastrocnemius muscle, perirenal, retroperitoneal and epididymal fat were isolated, fat/lean ratio was calculated and serum adiponectin and resistin were assayed. S did not affect lean body mass in any group. HFD was associated with elevated fat/lean ratio regardless of S administration. S at 10mg/Kg decreased fat/lean ratio in the HCD and HPD and adiponectin in the HFD group. S did not affect resistin in any group. Adiponectin was paradoxically elevated in the HFDS10 compared to the HCD or HPD S10 groups. Resistin was lower in the HCD compared to the HPD and HFD groups. Results suggest a preferential effect of S on body fat. The detrimental effect of S on adiponectin can be attributed to its sympathomimetic properties. Adiponectin was paradoxically elevated in the HFD and resistin in the HPD group, results that require further investigation.

Effect of isocaloric diets and sibutramine on food intake, body mass variation and serum TNF-alpha levels in rats.

BACKGROUND/AIM: The effect of isocaloric diets and sibutramine on dietary behaviour and TNF-alpha is poorly understood. The aim of the study was to investigate the effects of isocaloric diets and sibutramine on food intake, body mass variation and serum TNF-alpha in free-feeding rats. METHODS: Three groups of male Wistar rats (n = 63) were fed a high-fat diet, high-carbohydrate diet or high-protein diet for 13 weeks. In the last 3 weeks, each group was divided into 3 subgroups. Each subgroup received sibutramine 5 mg/kg, sibutramine 10 mg/kg or vehicle. Food intake was measured daily during the last week of the experiment; serum TNF-alpha was assayed and the body weight increasing rate was calculated. RESULTS: The high-fat diet was associated with increased food intake, a greater weight gain ratio and increased TNF-alpha levels. Sibutramine treatment did not affect the dietary behaviour of high-protein- or high-carbohydrate-fed rats, while it significantly attenuated the daily food intake and body weight gain rate in the high-fat group, at the dose of 10 mg/kg. TNF-alpha levels were not affected by sibutramine. CONCLUSIONS: High-fat feeding was associated with an increase in daily food intake, TNF-alpha levels and body weight gain rate, as well as with enhanced responsiveness to the anorectic effects of sibutramine. However, sibutramine did not affect TNF-alpha.

Coagulating intermittent cutting. Improved high-frequency surgery in transurethral prostatectomy.

OBJECTIVES: Transurethral prostatectomy (TURP) is the gold standard in surgical therapy of symptomatic bladder outlet obstruction. TURP is characterised by immediate treatment success, due to the removal of obstruction combined with a long-lasting improvement of symptoms and voiding parameters. In spite of good long-term results of TURP, intraoperative blood loss produces morbidity. We investigated a blood-sparing cut using a new high-frequency technology. METHODS: (1) A standard high frequency generator was extended in its function by additional electronics. (2) The possibility of a blood-sparing cut using 'coagulating intermittent cutting' (CIC cocut BMP) was quantified ex vivo using a blood-perfused porcine kidney. Four cuts next to each other were performed through the parenchyma using a standard resectoscope with a standard loop. This was done with a commercially available generator and CIC cocut BMP. The blood loss was determined semiqantitatively. RESULTS: (1) In a first step 'coagulating cutting' with coagulating and cutting periods (10/94-- 08/96) was developed. During each cut, phases with predominant cutting effect alternate with coagulating phases of defined duration. As a disadvantage, operation time increased due to lower cutting speed. In a second step cutting combined with coagulation effect with high voltage pulses -- 'coagulating intermittent cutting' (08/96--06/97) -- was developed. In this technique, the output signal consists of a pulse-modulated sinusoidal voltage with high amplitudes. But gas bubbles impaired vision. This finally resulted in the 'coagulating intermittent cutting' with constant voltage pulses and control of pulse intervals (CIC cocut BMP, since 07/97 up to now). (2) Comparing the function of the high-frequency generators in vitro, the Wilcoxon test for paired samples revealed a significant reduction of the observed bleeding with the CIC cocut BMP (p = 0.002). CONCLUSIONS: 'Coagulating intermittent cutting' improves the gold standard of TURP with reduced blood loss. The procedure is feasible with a standard resection equipment. The already trained surgeon has no further learning cure, and teaching of classical TURP is maintained.

Lethal dose curve: should its determination be made in healthy or in animal models of disease?

The lethal dose curve of phenylbutazone was assayed in control animals (A) and in animals in which an experimental arthritis had been produced by injection of Feund's complete adjuvant (B). Thirty male Wistar rats 3 months old were divided into two groups of 15 rats each: group A consisted of the control group and group B the adjuvant treated group. Arthritis was produced by a single injection of 0.15 Freund's complete adjuvant on the right footpad. The control group was injected with the adjuvant vehicle at the same site. Injections of 200 mg/kg of phenylbutazone once daily for ten days started fifteen days after administration of Freund's adjuvant. After the first injection of phenylbutazone the LD6.6 in group A corresponded to LD53 in group B while after the second injection the LD46.6 for the control group corresponded to LD100 in group B. We assume that the therapeutic index would be more representative if lethal dose curves were evaluated in animals suffering from a disease that the drug in question has been designated for.

Determination of glutamate by high-performance liquid chromatography with fluorescence detection in superfusates of rat brain preparations.

A method is described for the determination of glutamate in superfusates of cortical or hippocampal rat brain slices. This method is based on the precolumn derivatization of amino acids with orthopthaldialdehyde and mercaptopropionic acid, separation by high-performance liquid chromatography, and detection by fluorescence. By adjusting the concentrations of reagents and with the minimum dilution of the sample, it was possible to reproducibly measure the Ca2+-dependent K+-evoked release of neurotransmitter glutamate in superfusates of brain slices.

Effects of [D-Ala1] peptide T-NH2 and HIV envelope glycoprotein gp120 on cyclic AMP dependent protein kinases in normal and psoriatic human fibroblasts.

In addition to acquired immunodeficiency syndrome (AIDS), persons infected with human immunodeficiency virus often develop cutaneous manifestations, including severe psoriasis. In previous studies, we have established that psoriatic fibroblasts and erythrocytes obtained from psoriatic patients exhibit decreased levels of cyclic adenosine monophosphate (cAMP) dependent protein kinase (PKA) activity and of 8-azido-[32P]cAMP binding to the RI and RII regulatory subunits of PKA. Because treatment of patients with peptide T (an octapeptide sequence found in the human immunodeficiency virus envelope glycoprotein gp120) has been observed to result in an improvement in the psoriatic condition, studies were initiated to determine if peptide T and gp120 protein treatment of normal and psoriatic human fibroblasts resulted in any changes in PKA. Exposure of psoriatic fibroblasts to peptide T resulted in a time (4 h to 6 d) and dose [10(-14)-10(-8) M] dependent increase in the levels of 8-azido-[32P]cAMP binding to the RI and RII regulatory subunits of PKA, along with a corresponding increase in PKA activity. Peptide T exhibited a biphasic dose dependent response, with maximal effects on PKA noted at 10(-12)M peptide T. Treatment of normal human fibroblasts with peptide T did not result in any change in PKA levels. Conversely, treatment of normal human fibroblasts for 18 h with gp120 protein [10(-13) M] resulted in a significant decrease in the levels of 8-azido-[32P]cAMP binding to RI and RII and in PKA activity. The presence of peptide T blocked this effect of the gp120 protein. These results indicate that peptide T and gp120 protein may inversely alter the intracellular levels of 8-azido-[32P]cAMP binding to RI and RII, and of PKA activity in susceptible cells. These observed changes in the cyclic AMP-PKA signaling pathway, a biochemical marker for psoriasis, may offer some mechanistic insight into the noted beneficial effects of peptide T treatment, including an improvement in psoriatic lesions.

Oxidative modulation of cyclic AMP-dependent protein kinase in human fibroblasts: possible role in psoriasis.

Previous studies have established that cyclic AMP-dependent protein kinase (PKA) activity, as well as 8-azido-[32P]-cAMP binding to the RI and RII regulatory subunits, are decreased in cells from psoriatic patients compared to cells from normal patients. Here we show that the exposure of normal human dermal fibroblasts in culture to hydrogen peroxide and to oxygen free-radical generating systems decreased PKA activity, as well as cyclic AMP binding to the RI and RII regulatory subunits, to levels similar to those observed with psoriatic fibroblasts. Likewise, treatment of normal cytosolic preparations of PKA, as well as purified bovine PKA II, in vitro with free radical generating systems also resulted in decreased PKA activity and 8-azido [32P]-cAMP binding to the RI and RII regulatory subunits. Further, treatment of psoriatic fibroblasts with free radical scavenging agents such as vitamins E and C, and mannitol, and also with superoxide dismutase, restored the ability of RI and RII to bind 8-azido-[32P]-cAMP toward normal levels. Western blot analysis showed that the protein levels of the RI and RII subunits are similar in normal and psoriatic fibroblasts, and that the amounts of RI and RII are not altered by treatment of the cells with free radical-generating systems. These results suggest that oxidative modification may serve as a mechanism to alter PKA activity in human cells, and that an altered oxidative state may be involved in mediating the decrease in PKA activity and cyclic AMP binding noted in cells from psoriatic patients.

Effect of low-dose methylprednisolone on calcium balance and bone composition in rats.

Glucocorticoid induced osteoporosis has been associated with high doses and it has been partially attributed to decreased absorption and to increased calcium excretion. The present study examined the effect of low but effective doses of methylprednisolone (MP) on calcium balance and on skeleton in rats. Total duration of the experiment 29 days. Thirty-one male Wistar rats (MP group) were injected with 4mg/kg body weight MP s.c. at the 1st, 11th and 20th day of the experiment and 28 rats (C group) were used as matched controls. The 1st, 11th and 20th day of the experiment rats were placed in individual metabolic cages for three days. Food and water consumption were measured at the 2nd and 3rd days after each injection; urine and faeces were collected at the same days for calcium estimation. Calcium intake and excretion after each injection was significantly lower in the MP group as compared to controls. A statistically significant positive correlation between calcium consumption and calcium excretion was found in both groups resulting in a negative final balance. Rats were killed the 29th day. Adrenal weight was statistically significant lower in MP group (p<0.001). Morphometric properties were evaluated for the right femur. No significant difference was found between the two groups. Mineral and calcium content was slightly increased in the MP group. According to these results, it seems that methylprednisolone while effective on HPA axis did not have any effect on calcium absorption and on bone calcium deposition in rats.

Corticotropin-releasing-hormone levels in pregnancy-induced hypertension.

UNLABELLED: High levels of corticotropin-releasing hormone (CRH) circulate in the plasma of pregnant women especially during the third trimester and even higher levels have been reported in abnormal pregnancies of various etiologies. One of these etiologies is pregnancy-induced hypertension (PIH). OBJECTIVE: To measure CRH blood levels with a sensitive method in a large number of pregnant women with PIH, starting from very early stages of gestation, and to compare them with those in normal controls. STUDY DESIGN: Venous blood was withdrawn from, (a) 10 healthy women aged 20-35 years, (b) 62 pregnant women with PIH (109 samples), mean age 29.1 years and (c) 75 healthy pregnant women (81 samples), mean age 28.5 years, used as matched controls. In pregnant women, blood collection started at the 10th week of gestation. In 14 women from group b and in 22 from group c blood was withdrawn during labor as well. CRH was assayed by RIA. RESULTS: Levels in non-pregnant women were between 19.0-40.6 pg/ml (28.37 +/- 2.53 pg/ml, mean +/- S.E.M.). In both groups of pregnant women there was a progressive increase in plasma CRH levels becoming quite sharp towards the end of gestation. Between 10 and 20 weeks, CRH (mean +/- S.E.M.) in PIH group was 69.3 +/- 3.2 pg/ml versus 41.6 +/- 2.4 pg/ml in matched controls, at 21-25 weeks 168.0 +/- 12.8 pg/ml versus 58.5 +/- 3.8 pg/ml, at 32-35 weeks 1378.5 +/- 61.4 pg/ml versus 298.3 +/- 16.9 pg/ml and at 38 weeks 2800.0 +/- 114.1 pg/ml versus 825.0 +/- 59.8 pg/ml. At term, CRH levels were 3784.0 +/- 197.3 pg/ml in PIH, versus 1386.0 +/- 101.8 pg/ml in normal pregnancy. Statistically, at every stage of gestation, CRH levels were highly significantly different in the PIH group (P < 0.0005). One hour postpartum there was a c. 60% decrease in plasma CRH levels in both b and c groups. In three women with pre-eclampsia who underwent premature labor due to a dead fetus around the 30th week, very high levels were noticed in sequential blood samples for 4-5 weeks prior to labor. CONCLUSIONS: (a) CRH levels in women with PIH are significantly higher compared to healthy pregnant women at any stage of gestation starting from week 10; (b) very high levels during pregnancy might be predictive of premature labor or fetal loss; and (c) CRH measurement might prove to be a helpful diagnostic tool in women with pregnancy-induced hypertension.

Contribution to the problem of hyperprolactinaemia: experience with 4,199 prolactin assays and 117 prolactinomas.

OBJECTIVE: To help define the normal and pathological serum prolactin concentration ranges in adult females. PATIENTS AND METHODS: Serum prolactin was assayed in a mixed population of 4,199 women divided into five groups: (A) 753 with normal menses consulting for infertility (control group); (B) 2,523 with menstrual disorders; (C) 519 with hirsutism and normal cycles; (D) 201 with galactorrhoea and normal cycles; (E) 203 with galactorrhoea and menstrual disorders. RESULTS: In each group, a separate subset (2%, 3%, 1%, 15%, and 42.3%, respectively) was distinguished, with scattered prolactin values above 30 ng/mL, in which 117 prolactinomas were found, with an incidence ranging from 11 to 57% in cases with prolactin of 31-49 ng/mL and from 65 to 87.5% in those with prolactin > or = 50 ng/mL. The frequency distribution of values from 1 to 30 ng/mL was strikingly similar in the 753 subjects of group A and the 2,523 patients of group B: 91% and 92.6%, respectively, between 1 and 15 ng/mL, and 5.5% and 4.7% from 16 to 20 ng/mL. Values > 20 ng/mL were found in 4.4% of the subjects in group A, 5.5% in group B, 5.1% in group C, 21.8% in group D, and 18.7% in group E. CONCLUSIONS: (a) Normal prolactin values can be considered to include 1-15 ng/mL and 16-20 ng/mL, the former corresponding to the vast majority of normal subjects or patients, and the latter being the extreme limit of normal, for which from a clinical point of view repetition of the assay and a follow-up is advocated. (b) Values between 21 and 30 are rare and could be considered as hyperprolactinemia necessitating further investigation. (c) Values between 31 and 49 are suspect, and values > or = 50 ng/mL are suggestive of the presence of a prolactinoma, warranting thorough investigation.

Incidence and characteristics of microprolactinomas (3-5 mm) in 4199 women assayed for prolactin.

Difficulties and controversies still exist in the diagnosis of small (3-5 mm) prolactinomas (micro-PRL-omas). In the present study serum prolactin (PRL) was assayed in 4199 women aged 14-43 years belonging to 4 groups: A: 753 women with normal cycles (NC) and infertility (control group), B: 2523 with menstrual disorders, C: 519 with NC and hirsutism, D: 404 with galactorrhoea. The distribution of PRL values from 1 to 30 ng/ml was almost similar in the subjects of group A, B and C. Within this range the vast majority of subjects (91%, 92.2% and 88% respectively in these 3 groups and 83% in group D) had PRL levels from 1 to 15 ng/ml and together with the proportion of subjects with PRL values 16 to 20 ng/ml they included 96.7% of the entire mixed population. A proportion of scattered outlying PRL values above 30 ng/ml was found in each group (A = 2%, B = 3%, C = 1% and D = 28.7%) and in this subset 117 prolactinomas (PRL-omas) were found, 19 (23%) in the 83 subjects with PRL levels 31-49 ng/ml and 98 (75.3%) in the 130 subjects with PRL values greater than or equal to 50 ng/ml. Of the 117 PRL-omas 9 were bigger than 10 mm and 4 had a size from 6 to 9 mm. In the remaining 104 the size was presumed from direct or indirect radiological evidence to be 3-5 mm.(ABSTRACT TRUNCATED AT 250 WORDS)

Retinoylation of the cAMP-binding regulatory subunits of type I and type II cAMP-dependent protein kinases in HL60 cells.

Retinoylation (retinoic acid acylation) is a post-translational modification of proteins occurring in a variety of eukaryotic cell lines. There are at least 20 retinoylated proteins in the human myeloid leukemia cell line HL60 (N. Takahashi and T.R. Breitman (1990) J. Biol. Chem. 265, 19, 158-19, 162). Here we found that some retinoylated proteins may be cAMP-binding proteins. Five proteins, covalently labeled by 8-azido-[32P]cAMP which specifically reacts with the regulatory subunits of cAMP-dependent protein kinase, comigrated on two-dimensional polyacrylamide gel electrophoresis with retinoylated proteins of Mr 37,000 (p37RA), 47,000 (p47RA), and 51,000 (p51RA) labeled by [3H]retinoic acid treatment of intact cells. Furthermore, p47RA coeluted on Mono Q anion exchange chromatography with the type I cAMP-dependent protein kinase holoenzyme and p51RA coeluted on Mono Q anion exchange chromatography with the type II cAMP-dependent protein kinase holoenzyme. An antiserum specific to RI, the cAMP-binding regulatory subunit of type I cAMP-dependent protein kinase, immunoprecipitated p47RA. An antiserum specific to RII, the cAMP-binding regulatory subunit of type II cAMP-dependent protein kinase, immunoprecipitated p51RA. These results indicate that both the RI and the RII regulatory subunits of cAMP-dependent protein kinase are retinoylated. Thus, an early event in RA-induced differentiation of HL60 cells may be the retinoylation of subpopulations of both RI and RII.

Hypothalamic and suprahypothalamic effects of prolonged treatment with dexamethasone in the rat.

Corticosteroid type I and II receptors mediate the negative feedback effects of these hormones at various central nervous system sites involved in the regulation of the hypothalamic-pituitary-adrenal (HPA) axis. To examine the effects of chronic treatment with dexamethasone (DEX), a type 2 receptor agonist, on the regulation of this axis, male Sprague-Dawley rats weighing 200-250 g were given daily injections of DEX for 1,2,3, and 4 weeks or were treated with a subcutaneously implanted DEX-releasing minipump for one week. At the end of treatment, the animals were weighed and brains and truncal blood were collected. Daily intermittent DEX treatment reduced the body weight of the rats in a time-dependent fashion, but had little or no effect on their wet brain weight. Plasma ACTH and corticosterone, measured by RIA, were fully suppressed after one week of intermittent treatment and did not show any further reduction in rats treated for longer periods. In these animals, the content of immunoreactive corticotropin-releasing hormone (iCRH), arginine vasopressin (AVP), ACTH and beta-endorphin (beta-EP) in the hypothalamus, hippocampus, cerebral cortex and cerebellum and pituitary ACTH content did not show any difference compared to vehicle-treated rats. In contrast, continuous DEX treatment increased iCRH content in the cortex, reduced AVP content in the cerebellum, increased ACTH content in the hippocampus, decreased ACTH and beta-EP content in the hypothalamus, and reduced pituitary ACTH content. Hypothalami explanted from rats treated with DEX for one week released lower basal amounts of iCRH in vitro and did not respond to a maximally stimulatory concentration of serotonin (5-HT), a known CRH secretagogue. Continuous DEX administration suppressed also potassium chloride-induced iCRH release. Interestingly, hypothalami explanted from rats receiving daily injection of vehicle, but not from unhandled, untreated controls, did not respond to 5-HT with an increase of iCRH release in vitro. In conclusion, prolonged and continuous, but not intermittent, administration of DEX had a strong effect on brain neuropeptide content. Both regimens of DEX reduced the hypothalamic iCRH responsiveness to stimuli in vitro. Chronic handling also decreased the responsiveness of the hypothalamus to a stimulatory neurotransmitter and may confound the interpretation of data pertinent to inhibitory mechanisms.

High chemotactic response to platelet-derived growth factor of a teratocarcinoma differentiated mesodermal cell line.

Evidence is presented that a differentiated mesodermal line (MES-1) from P19 EC cells express a high chemotactic response to platelet-derived growth factor (PDGF) as assayed in a blind-well modified Boyden chamber. Compared to the NIH 3T3 fibroblasts the chemotactic response of MES-1 is increased by 10-fold at 0.3 ng/ml of PDGF, 4-fold at 1.25 ng/ml of PDGF, 2-fold at 2.5 ng/ml of PDGF. In contrast, PDGF induces the same increase in [3H]thymidine incorporation in both cell lines, made quiescent under reduced serum concentration. This high chemotactic response to PDGF seems specific for these mesodermal cells. Among the different teratocarcinoma cells tested, including stem cells (F9, PC 13, PCC4) and endodermal derivatives (PYS, F9 with retinoic acid, PSA 5E), only the visceral endodermlike cells (PSA5E) are slightly attracted by PDGF. This chemotactic response to PDGF is not related to the presence or characteristics of the type B PDGF receptors, which are less numerous in MES-1 cells (10(5) receptors/cell, KDa 1,2 mM) compared to NIH 3T3 cells (64 X 10(4) receptors per cell, KDa 1,8 nM). The MES-1 cell line might be of interest for studying the chemotactic effect of PDGF. These results also suggest a role for this soluble factor in cell migration during early embryogenesis.

Glucocorticoids inhibit estradiol-mediated uterine growth: possible role of the uterine estradiol receptor.

Stress-related activation of the hypothalamic-pituitary-adrenal axis (HPA) is associated with suppression of the reproductive axis. This effect has been explained by findings indicating that corticotropin-releasing hormone suppresses hypothalamic gonadotropin-releasing hormone (GnRH) secretion via an opioid peptide-mediated mechanism, and that glucocorticoids suppress both GnRH and gonadotropin secretion and inhibit testosterone and estradiol production by the testis and ovary, respectively. To evaluate whether glucocorticoids suppress the effects of estradiol on its target tissues, we examined the ability of dexamethasone to inhibit estradiol-stimulated uterine and thymic growth in ovariectomized rats. Estradiol alone, given daily for 5 days, caused dose-dependent uterine and thymic growth. Dexamethasone alone, given daily for 5 days, caused a dose-dependent decrease in body weight gain and in thymic growth. When estradiol and dexamethasone were administered simultaneously, however, body weight gain and thymic growth were also inhibited (p less than 0.05). Dexamethasone decreased estradiol-induced uterine cytosolic and nuclear estrogen receptor concentrations (E2 R0, p less than 0.05; E2nR0, respectively), but had no effect on estradiol-induced progesterone receptor concentrations (P4R0, p greater than 0.05). Levels of uterine glucocorticoid receptors were not affected by estrogen and/or dexamethasone treatment. These findings suggest that stress levels of glucocorticoids, administered over a 5-day interval, block the estradiol-stimulated growth of female sex hormone target tissues. This effect may be partially mediated by a glucocorticoid-induced decrease of the estradiol receptor concentration. Thus, another mechanism by which the HPA may influence reproductive function during stress is by a direct effect of glucocorticoids on the target tissues of sex steroids.

Effect of retinoic acid on cAMP dependent protein phosphorylation in psoriatic fibroblasts.

Retinoic acid treatment of psoriatic fibroblasts increases the activity of cyclic AMP dependent protein kinase. In this study we report that retinoic acid treatment of cultured psoriatic fibroblasts modifies their subsequent cAMP dependent protein phosphorylation. In the soluble fraction of normal fibroblasts cAMP clearly enhances the in vitro phosphorylation of proteins of MW 37,49,54,56,68,83 kD while retinoic acid treatment of the same cells results in a decrease of the cAMP dependent phosphorylation of the first five of the same proteins. In contrast, in psoriatic fibroblasts from psoriatic patients retinoic acid either has no effect or increases the cAMP dependent phosphorylation of some of these proteins. Moreover the phosphorylation of a protein of MW 54 kD, undetectable in untreated psoriatic cells, is more phosphorylated in the presence of cAMP after retinoic acid treatment. The appearance of this phosphorylated proteins is time dependent and dose dependent upon the addition of retinoic acid. These in vitro phosphorylation results suggest that retinoic acid treatment of psoriatic fibroblasts change the level of cAMP dependent phosphorylation of some cytosolic proteins. These specific phosphorylations could be implicated in a variation of cell functions.

Altered hormonal responses: markers for embryonal and embryonic carcinoma stem cells and their differentiated derivatives.

Teratocarcinoma cells in culture offer an in vitro system for studying certain aspects of embryonic differentiation. To gain some insight into regulatory systems that might be operative during early development, we have characterized the alterations that occur in the hormonal responsiveness of the membrane adenylate cyclase of different embryonal carcinoma cell lines with differentiation. Each undifferentiated embryonal carcinoma stem cell studied (F9, PCC4, PC13, P19) has an adenylate cyclase system predominantly activated by calcitonin. Of great interest is the fact that cAMP production is also enhanced specifically by calcitonin in an embryo-derived stem cell line. Differentiation of the embryonal carcinoma stem cell population toward parietal endoderm results in a decrease in calcitonin activation with a concomitant appearance of sensitivity to parathyroid hormone. Differentiation toward visceral endoderm is characterized by a lack of response of the adenylate cyclase system to both calcitonin and parathyroid hormone. These results indicate that the changes noted in adenylate cyclase hormonal responsiveness might serve as useful markers during early stages of differentiation.

Diagnosis of ovarian follicular cysts from birth to puberty: a report of twenty cases.

Twenty girls aged 1 day to 17 years have been studied for ovarian follicular cysts. Clinical features leading to the discovery of the follicular cyst were different in prepubertal girls and in girls whose cyst was discovered during puberty. Before seven years of age, four girls presented a precocious pseudopuberty where breast development was in contrast with very low pituitary gonadotropin levels; two girls in this age group were diagnosed after complaining about abdominal pain. In two cases the diagnosis was prenatal during routine ultrasonography of the mother. After ten years of age, abnormal menses (5 cases) or acute abdominal pain (5 cases) were the main clinical features. In only one case the cyst presented as an abdominal mass. Follow-up of the 20 patients showed: spontaneous disappearance of the cyst within 3 to 32 weeks in 9 cases; ovariectomy in 8 cases, due to a torsion of a large cyst (over 55 mm) in 7 children and because of the fear of a possible tumor in one; limited resection of the cyst in 4 cases. By systematic ultrasonography, discovery of an ovarian cyst as defined by a non-echogenic area over 20 mm may occur relatively often in young girls. Spontaneous disappearance is frequent when the cyst is small (under 55 mm). Torsion of large cysts remains the major complication.

Comparison of growth hormone response to growth hormone-releasing factor 1-44 according to the combined study of sleep secretion with the responses to pharmacologic stimuli.

The growth hormone (GH) response to GH-releasing factor (GRF) was studied in 54 severely growth-retarded patients (-2.1 to -6.5 SD) aged from 5 to 20 years (32 males and 22 females), among whom 34 were prepubertal and 20 at early pubertal stages. The patients were also submitted to a standard evaluation of their GH secretion, consisting of at least two classical pharmacologic stimulation (CPS) tests, such as ornithine, arginine and/or insulin, and one study of the GH sleep secretion (SS). The results of the standard evaluation allowed to distinguish 5 groups: (I) endocrinologically normal (n = 26); (II) completely GH deficient (n = 5); (III) partially GH deficient (n = 8); (IV) dissociated GH secretions with normal SS (n = 9), and (V) dissociated GH secretions with low SS (n = 6). The GH responses to GRF were correlated with both responses to CPS and SS. There was a large overlap of the individual responses to GRF between the 5 groups, but the mean responses in groups II, IV and V were significantly lower than in group I. Furthermore, the mean responses of groups IV and V were in the lower range of the normal. It is concluded that the GRF test may be useful to ascertain the diagnosis of functional or partial GH deficiency when the responses to CPS and SS are dissociated.