Differences in bisphenol A and estrogen levels in the plasma and seminal plasma of men with different degrees of infertility.

The general population is potentially exposed to many chemicals that can affect the endocrine system. These substances are called endocrine disruptors (EDs), and among them bisphenol A (BPA) is one of the most widely used and well studied. Nonetheless, there are still no data on simultaneous measurements of various EDs along with steroids directly in the seminal fluid, where deleterious effects of EDs on spermatogenesis and steroidogenesis are assumed. We determined levels of BPA and 3 estrogens using LC-MS/MS in the plasma and seminal plasma of 174 men with different degrees of infertility. These men were divided according their spermiogram values into 4 groups: (1) healthy men, and (2) slightly, (3) moderate, and (4) severely infertile men. Estradiol levels differed across the groups and body fluids. Slightly infertile men have significantly higher BPA plasma and seminal plasma levels in comparison with healthy men (p<0.05 and p<0.01, respectively). Furthermore, seminal BPA, but not plasma BPA, was negatively associated with sperm concentration and total sperm count (-0.27; p<0.001 and -0.24; p<0.01, respectively). These findings point to the importance of seminal plasma in BPA research. Overall, a disruption of estrogen metabolism was observed together with a weak but significant impact of BPA on sperm count and concentration.

Determination of homocysteine in cerebrospinal fluid as an indicator for surgery treatment in patients with hydrocefalus.

Increased homocysteine levels in serum are typical features of neurodegenerative brain diseases including hydrocephalus. The most frequent therapeutic approach consists of the insertion of a shunt, connecting the brain ventricles to an alternative drainage site. To decide whether the patient should undergo this, the lumbar drainage test is usually carried out to distinguish patients who can benefit from the shunt insertion. In searching for other potential biochemical markers for shunt indication we determined homocysteine levels in CSF during the lumbar drainage test. Homocysteine in CSF was measured during the 5-day lumbar drainage test in 27 patients with normal-pressure hydrocephalus (NPH) and in 25 patients with excluded hydrocephalus. A novelized gas chromatography method with flame ionization detection (GC-FID) was developed and evaluated. During the first two days of lumbar drainage, the levels of CSF homocysteine in NPH patients were significantly higher compared to the controls, while on the fifth day, the homocysteine levels in patients with hydrocephalus reached the level of controls. Determination of CSF homocysteine in patients with confirmed or suspected hydrocephalus may serve as an independent marker for deciding on their further treatment strategy.

[Phytoestrogenes in menopause: working mechanisms and clinical results in 28 patients]. / Fytoestrogeny v menopauze: mechanismus úcinku a výsledky lécby v sestave 28 pacientek.

OBJECTIVE: Comparison of phytoestrogen treatment efficacy in menopausal women with and without ability to metabolise phytoestrogens. DESIGN: Clinical trial. SETTING: Department of Obstetrics and Gynaecology, Regional Hospital, Mlada Boleslav. METHODS: 28 menopausal women were treated with phytoestrogens in dose 80 mg daily. Before start and after finishing of treatment urinary concentrations of active metabolite S-equol were measured using ELISA method. Similarly before and after treatment Kupperman's index was measured. Patients with urinary concentrations of S-equol above 1 ng/ml were considered as S-equol producers, remaining patients formed control group. RESULTS: 16 out of 28 women were considered as S-equol producers, remainig 12 as a non-producers. Initial urinary concentrations of S-equol were 0.34 +/- 0.37 ng/ml in producers group and 0.29 +/- 0.30 ng/ml in non-producers. After finishing of therapy urinary concentration of S-equol increased to 10.67 +/- 11.57 ng/ml (p = 0.002) in producers group and 0.34 +/- 0.30 ng/ml (p = 0.701) in non-producers. Kupperman's index values were 23.44 +/- 11.57 in producers group and 17.25 +/- 7.78 in non-producers. After therapy value of Kupperman's index decreased to 14.44 +/- 9.97 (p = 0.003) in producers and to 12.00 +/- 7.18 (p = 0.100) in non-producers. No correlation between improvement in Kupperman's index and urinary concentration of S-equol after therapy was found similarly as between urinary concentration of S-equol before and after therapy in producents group. CONCLUSION: Significant phytoestrogen treatment effect in menopausal women producing S-equol was proven. Testing method for S-equol production introduced by our team togehter with suggested threshold urine concentration level of 1 ng/ml allows precise distinction of producers and non-producers of S-equol and subsequently to predict better treatment effect of phytoestrogens.

Hyperandrogenic states in pregnancy.

Hyperandrogenic states in pregnancy are almost always the result of a condition that arises during pregnancy. The onset of virilization symptoms is often very fast. The mother is protected against hyperandrogenism by a high level of SHBG, by placental aromatase and a high level of progesterone. The fetus is protected from the mother's hyperandrogenism partly by the placental aromatase, that transforms the androgens into estrogens, and partly by SHGB. Nevertheless there is a significant risk of virilization of the female fetus if the mother's hyperandrogenic state is serious. The most frequent cause of hyperandrogenic states during pregnancy are pregnancy luteoma and hyperreactio luteinalis. Hormonal production is evident in a third of all luteomas, which corresponds to virilization in 25-35 % of mothers with luteoma. The female fetus is afflicted with virilization with two thirds of virilized mothers. Hyperreactio luteinalis is created in connection with a high level of hCG, e.g. during multi-fetus pregnancies. This condition most frequently arises in the third trimester, virilization of the mother occurs in a third of cases. Virilization of the fetus has not yet been described. The most serious cause of hyperandrogenism is represented by ovarian tumors, which are fortunately rare.

TRH-like peptides.

TRH-like peptides are characterized by substitution of basic amino acid histidine (related to authentic TRH) with neutral or acidic amino acid, like glutamic acid, phenylalanine, glutamine, tyrosine, leucin, valin, aspartic acid and asparagine. The presence of extrahypothalamic TRH-like peptides was reported in peripheral tissues including gastrointestinal tract, placenta, neural tissues, male reproductive system and certain endocrine tissues. Work deals with the biological function of TRH-like peptides in different parts of organisms where various mechanisms may serve for realisation of biological function of TRH-like peptides as negative feedback to the pituitary exerted by the TRH-like peptides, the role of pEEPam such as fertilization-promoting peptide, the mechanism influencing the proliferative ability of prostatic tissues, the neuroprotective and antidepressant function of TRH-like peptides in brain and the regulation of thyroid status by TRH-like peptides.

Roles of steroid sulfatase in brain and other tissues.

Steroid sulfatase (EC 3.1.6.2) is an important enzyme involved in steroid hormone metabolism. It catalyzes the hydrolysis of steroid sulfates into their unconjugated forms. This action rapidly changes their physiological and biochemical properties, especially in brain and neural tissue. As a result, any imbalance in steroid sulfatase activity may remarkably influence physiological levels of active steroid hormones with serious consequences. Despite that the structure of the enzyme has been completely resolved there is still not enough information about the regulation of its expression and action in various tissues. In the past few years research into the enzyme properties and regulations has been strongly driven by the discovery of its putative role in the indirect stimulation of the growth of hormone-dependent tumors of the breast and prostate.

Homocysteine during the menstrual cycle in depressive women.

BACKGROUND: Two possible factors that may have a causal relation with both depressive disorder and cardiovascular disease are elevated homocysteine and steroid hormones. Our previous study found significant changes in the plasma homocysteine concentration during the menstrual cycle in healthy women. The purpose of this study therefore was to test homocysteine in depressive women treated with fluoxetine during the menstrual cycle. MATERIALS AND METHODS: Thirteen premenopausal women suffering from mixed anxiety-depressive disorder and a control group of 15 healthy women were enrolled in this study. The homocysteine concentration was determined by high-performance liquid chromatography with fluorescence detection, and estradiol, progesterone and cortisol by RIA methods. RESULTS: We found significantly higher plasma homocysteine concentrations in the follicular phase than in the luteal phase of the menstrual cycle in both the depressive group (P < 0.003) and the controls (P < 0.0009). Moreover, the patient values of total homocysteine were significantly higher in the follicular phase (P < 0.03) and also in the luteal phase (P < 0.007) than the values of the controls. Estradiol and cortisol were significantly higher in the follicular phase of the patients compared with the control group. CONCLUSION: According to our results, women suffering from mixed anxiety-depressive disorder have not only significantly different concentrations of homocysteine in the follicular and luteal phase of the menstrual cycle but also higher plasma homocysteine compared with healthy women. More elevated homocysteine in the depressive than in the healthy premenopausal women points to the notion that psychological factors might be important when considering the homocysteine concentration.

Homocysteine and steroids levels in metformin treated women with polycystic ovary syndrome.

UNLABELLED: Risk for atherosclerosis is increased in women with polycystic ovary syndrome (PCOS). Homocysteine (Hcy) is one of the independent risk factors for ischemic heart disease. We examined the effect of metformin (M) treatment on Hcy levels, steroids and glycide tolerance in PCOS. MATERIAL AND METHODS: 9 women with PCOS (defined as hyperandrogenemia and chronic anovulation); age 20 +/- 3.8 yrs, BMI 28.1 +/- 6.5 kg/m(2); examined in the follicular phase of spontaneous menstrual cycle before and after 27 +/- 4 weeks of treatment with M 1000 mg/day. The plasma concentrations of Hcy, DHEA, DHEA-S, cortisol (F), allopregnanolone (HPO), 17OHpregnenolone (17OHPl), insulin (I) and blood glucose (G) before and after the course of M were measured. RESULTS: After the course of M, Hcy significantly increased (10.1 +/- 2.6 to 13.4 +/- 5.1 micromol/l, p < 0.05.). There was no significant change in levels of I, HPO, F, DHEA-S and 17OHPl. DHEA levels increased significantly (from 26.9 +/- 15.7 to 44.4 +/- 24.6 nmol/l, p < 0.05). A borderline significant trend towards reduction in waist-hip ratio was seen (from 0.986 +/- 0.042 to 0.951 +/- 0.085; p < 0.06). CONCLUSIONS: Treatment with metformin in women with PCOS can lead to the increase in homocysteine levels--a risk factor for atherosclerosis.

Effect of treatment of hypothyroidism on the plasma concentrations of neuroactive steroids and homocysteine.

Autoimmune thyroiditis with hypothyroidism is frequently accompanied by symptoms of psychiatric disorders and atherogenic changes in lipid metabolism. Recent studies suggest that some neuroactive steroids and homocysteine are involved in the pathophysiology of both disorders. Homocysteine metabolism may be affected by some steroids. We were interested if the treatment of hypothyroidism would affect the above factors. We studied plasma concentrations of allopregnanolone, pregnenolone sulfate, dehydroepiandosterone and its sulfate, progesterone, estradiol and homocysteine in 14 patients (12 women, 2 men) during the 3-month treatment with levothyroxine. Steroids and thyroid function were monitored by measuring thyrotropin, free triiodothyronine, free thyroxine and levels of thyroid antimicrosomal antibodies and antibodies to thyroglobulin. We have found that with the restoration of the thyrotropin level, free triiodothyronine, free thyroxine and homocysteine levels decreased, but the levels of steroids were not significantly altered. Steroid concentrations correlated negatively with the level of thyroid antimicrosomal antibodies.

Homocysteine in breast cyst fluid.

BACKGROUND: Elevated plasma homocysteine concentrations have been reported in a variety of carcinoma, including those of the breast. The risk of breast cancer is higher in patients suffering from gross cystic disease. The breast cyst fluid contains unusual amounts of low- and high- molecular substances, including steroid hormones and their conjugates. The present study was undertaken to find out the presence of homocysteine in the fluid filling the cysts and have its concentration compared with other thiols, levels of Na+/K+ ratio and steroid hormones. Materials and methods Fourteen women suffering from gross cystic disease were enrolled in this study. Cystic concentrations of homocysteine (Hcy), cysteine (Cys), cysteinylglycine (Cys-Gly) and glutathione (GSH) were determined by high performance liquid chromatography, with fluorescence detection; estradiol (E2), progesterone, allopregnanolone and pregnenolone sulfate (PregS) by RIA methods. RESULTS: Mean levels of Hcy, Cys, Cys-Gly, Na+/K+, E2 and PregS in the fluid filling the breast cysts were significantly higher than the corresponding plasma concentrations. In addition, a negative correlation was found between cystic Hcy and the Na+/K+ ratio (Rs = -0.72, P = 0.003) and positive correlations between cyst Hcy and estradiol (Rs = 0.64, P = 0.018) and Hcy and PregS (Rs = 0.60, P = 0.025). Conclusion The study provides the first evidence of thiol concentrations in the breast cyst fluid. The finding of a negative correlation between homocysteine and the Na+/K+ ratio support the idea that the homocysteine concentration in breast cysts might be used clinically as a marker for the development of breast cancer disease.

Allopregnanolone, pregnenolone sulfate, and epitestosterone in breast cyst fluid.

The risk of breast cancer is 2 to 5 times higher in patients suffering from gross cystic disease. Breast cysts are categorized into two groups (type I and type II) according to the concentration of electrolytes in the cyst fluid. The two types also differ with respect to accumulation of steroids and steroidogenic enzyme activity. In type I cysts a higher risk of breast carcinoma could be expected. Here, we studied a possible relationship between the type of cyst and levels of epitestosterone (an endogenous antiandrogen), allopregnanolone (a product of 5alpha-reductase activity), and pregnenolone-sulfate (an activator of N-methyl-D-asparate receptors). We have found five times higher levels of epitestosterone in BCF in comparison with the circulation. Allopregnanolone levels were similar to those in plasma of women in the luteal phase of the menstrual cycle. Pregnenolone-sulfate levels in BCF were about two orders of magnitude higher when compared with the circulation. No differences were found in concentrations of the steroids studied between the types of cysts.

The effect of epitestosterone on estrogen biosynthesis in vitro.

OBJECTIVE: The concentrations of epitestosterone in human serum correlates negatively with that of estradiol. The possible explanation of this relation was addressed, and the influence of epitestosterone on kinetics of estradiol formation in vitro was evaluated. METHODS: The concentration of epitestosterone was measured in serum of 54 men participating in a screening program for prostate disease. Epitestosterone inhibition of aromatase and 17beta-hydroxysteroid dehydrogenase activities was tested in vitro in the system consisting of human placental microsomes, NADPH or NAD and NADP respectively, and epitestosterone in increasing concentrations. Testosterone, androstenedione, estrone and 17beta-estradiol were utilized as substrates. RESULTS: A significant negative correlation between epitestosterone and estradiol levels in human male serum was found. No inhibition of aromatase activity was observed; however, inhibition of 17beta-hydroxysteroid dehydrogenase was found preferentially in the direction leading to oxidation of the C-17 hydroxy group. The inhibitory effect of epitestosterone was more pronounced with androgens as substrates. CONCLUSION: Epitestosterone could influence the formation of estradiol in vitro rather by inhibition of 17beta-hydroxysteroid dehydrogenase than by blocking aromatase activity.

Changes of plasma total homocysteine levels during the menstrual cycle.

BACKGROUND: It is known that plasma total homocysteine (tHcy) levels are lower in premenopausal and pregnant women compared with postmenopausal women. To confirm the suggestion that sex steroid hormones are nongenetic factors affecting homocysteine metabolism, we investigated the effect of natural steroid hormone levels on the fasting plasma tHcy in healthy women during the menstrual cycle. MATERIALS AND METHODS: Fifteen premenopausal women were enrolled in this study. Plasma tHcy, estradiol, progesterone and cortisol concentrations were measured in the luteal and follicular phase. The plasma tHcy concentration was determined by high performance liquid chromatography with fluorescence detection, and the steroid hormones by RIA methods. RESULTS: Mean homocysteine values increased from 7.8 micromol L-1 in the luteal phase to 8.9 micromol L-1 in the follicular phase (P < 0.000005, Student's paired t-test). We also found slight negative but insignificant correlations of homocysteine levels with estradiol in both phases of the menstrual cycle. In the case of cortisol and progesterone, no significant correlations with plasma homocysteine were found. CONCLUSION: The study provides the first evidence of significant differences in plasma homocysteine concentration during the menstrual cycle. From our observed findings it is necessary to account for the phase of the menstrual cycle when determining homocysteine in premenopausal women.

Allopregnanolone in women with premenstrual syndrome.

5Alpha-reduced metabolites of progesterone, especially 3alpha-hydroxy-5alpha-pregnan-20-one (allopregnanolone) are now listed to neurosteroids. Their anesthetic properties can be explained by their allosteric interaction with GABA(A) receptors. A woman's organism is provided with an abundance of progesterone during the luteal phase of the menstrual cycle and its level falls down sharply with the first day of menses. The level of allopregnanolone follows that of progesterone. Many women suffer from so called premenstrual syndrome (PMS) during the luteal phase. In this study we have determined allopregnanolone and progesterone levels and their ratios in order to assess the over-all activity of C21-steroid 5alpha-reductase in these patients and in controls. Significantly lower levels of both progesterone and allopregnanolone than in controls have been found in PMS patients in the follicular phase only.

A novel radioimmunoassay of allopregnanolone.

A radioimmunoassay for determination of 3 alpha-hydroxy-5 alpha-pregnan-20-one (allopregnanolone) in serum or plasma has been developed and evaluated. The method employs rabbit antiserum to 3 alpha-hydroxy-5 alpha-pregnane-11,20-dione-11-O-carboxymethyloxime bovine serum-albumin conjugate and tritiated radioligand. The main cross-reactant interfering in the assay, progesterone, is eliminated by permanganate oxidation. Two assay variants were compared, with and without a micro-column chromatography. The simplified variant appeared to be reliable enough for determination of allopregnanolone in normally menstruating women at luteal phase, whereas the column-chromatography step is necessary when analyzing samples of expected low analyte concentration as in women in follicular phase, postmenopausal women, or in men. The levels of allopregnanolone in healthy women correlated excellently with progesterone in agreement with previous findings.

The effect of epitestosterone on gonadotrophin synthesis and secretion.

The effects of 3-week treatment with increasing doses of epitestosterone (ET) on gonadotrophin gene expression and secretion, on testosterone and 5 alpha-dihydrotestosterone (DHT) levels, and on the weight of testes and prostates, were studied in intact adult male rats. The hormones were delivered by means of silastic capsules of different lengths filled with the steroid. One group of rats received testosterone (T) instead of ET, to compare the results with previous studies concerning the testosterone effect. The controls were given capsules with glucose only. Treatment with ET, as well as with T, significantly reduced the weights of prostates. When the data from ET-treated rats and controls were combined, a significant negative correlation (P < 0.001) was found between the weight of prostates and serum ET. T, in contrast to ET, also decreased significantly the weights of testes, ET treatment caused a significant reduction of serum T levels but only an insignificant decline of DHT levels, independent of the dose. Serum and pituitary (p) luteinizing hormone (LH) levels in the ET-treated rats did not change. Pituitary mRNA contents for the beta LH subunit (beta LH-mRNA) showed a dose-dependent significant increase, up to 170% (P < 0.01), with ET treatment. pFSH decreased with the lowest ET (2 cm) dose (P < 0.05), but no change was observed with the other doses. The mRNA for the common alpha-subunit also increased with the ET load. In conclusion, ET acts at several sites in the regulation of gonadotrophin formation and release. It enhances the steady-state mRNA levels of both gonadotrophins in the pituitary.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of epitestosterone on the plasma levels of LH and FSH in ovariectomized immature rats.

Immature ovariectomized female rats primed with estradiol or without estrogen priming were treated with epitestosterone i.p. After 7 h blood was collected and LH and FSH levels were determined. The dose-response relationship was a biphasic one. LH and less markedly FSH levels decreased under epitestosterone treatment with doses up to 10 mg, whereas at higher doses an increase of gonadotrophins was observed.

Estrogen receptors in male gynecomastia.

Nineteen tissue specimens obtained at surgery of male mammary gland due to persisting gynecomastia were assessed for the presence of estrogen receptors. In 13 cases the receptors were detected with the use of charcoal-dextran technique and 3H-estradiol as a ligand, the detection limit of such method being 0.2 fmol/mg of total protein. The average number of estrogen binding sites (mean +/- S. D.) was 2.49 +/- 2.47 fmol/mg total protein with the apparent association constant 2.8 X 10(9) 1 mol-1. The specificity of ligand binding to the receptor was confirmed by gradient centrifugation.