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PubMed2002

Effects of EDTA and Sodium Citrate on hormone measurements by fluorometric (FIA) and immunofluorometric (IFMA) methods.

BACKGROUND: Measurements of hormonal concentrations by immunoassays using fluorescent tracer substance (Eu3+) are susceptible to the action of chemical agents that may cause alterations in its original structure. Our goal was to verify the effect of two types of anticoagulants in the hormone assays performed by fluorometric (FIA) or immunofluorometric (IFMA) methods. METHODS: Blood samples were obtained from 30 outpatients and were drawn in EDTA, sodium citrate, and serum separation Vacutaine&rcircledR;Blood Collection Tubes. Samples were analyzed in automatized equipment AutoDelfiatrade mark (Perkin Elmer Brazil, Wallac, Finland) for the following hormones: Luteinizing hormone (LH), Follicle stimulating homone (FSH), prolactin (PRL), growth hormone (GH), Sex hormone binding globulin (SHBG), thyroid stimulating hormone (TSH), insulin, C peptide, total T3, total T4, free T4, estradiol, progesterone, testosterone, and cortisol. Statistical analysis was carried out by Kruskal-Wallis method and Dunn's test. RESULTS: No significant differences were seen between samples for LH, FSH, PRL and free T4. Results from GH, TSH, insulin, C peptide, SHBG, total T3, total T4, estradiol, testosterone, cortisol, and progesterone were significant different between serum and EDTA-treated samples groups. Differences were also identified between serum and sodium citrate-treated samples in the analysis for TSH, insulin, total T3, estradiol, testosterone and progesterone. CONCLUSIONS: We conclude that the hormonal analysis carried through by FIA or IFMA are susceptible to the effects of anticoagulants in the biological material collected that vary depending on the type of assay.

Journal Article↗
PubMed1985

Analysis of computer-generated hydropathy profiles for human glycoprotein and lactogenic hormones.

Analysis of hydrophobic and hydrophilic regions of human lactogenic (PRL, GH, and placental lactogen) and glycoprotein hormones (FSH, LH, TSH, and hCG) by the method of Kyte and Doolittle has been performed. A BASIC program, developed for the IBM personal computer, produces graphical and tabular results. The net hydropathy value, a new parameter based on the Kyte and Doolittle analysis which may be useful for comparing various polypeptides, was developed. This value is correlated with physical properties, such as solubility of the glycoprotein hormones. New (more hydrophilic) indices were assigned for glycosylated asparagine, serine, and threonine residues, and slightly more hydrophilic indices were assigned for half-cystines found in disulfide bonds. The results indicate that the so-called determinant loop of Ward and Moore is hydrophilic (accessible) and suggest that regions on either side of this loop should also be considered as potential effectors of hormone specificity. It is suggested that binding (protein-protein interaction) sites tend to be modestly hydrophilic, but also contain residues that could interact through the hydrophobic effect.

Chemical Phenomena↗
PubMed1978

Detection of ovulation by a radioreceptor assay for human luteinizing hormone.

A specific, sensitive, and rapid radioreceptor assay (RRA), employing membranes from bovine testes as receptor and [125I]hLH as radioligand, has been developed for measurement of human LH in serum. This RRA was used to determine the time of ovulation in seven women. For comparison, four hourly values around midcycle were measured by RIA. The sensitivity of the RRA was 0.78 ng/ml serum and could be increased by prolonged incubation. The coefficient of within and between assay variation at the 50% inhibition level was 7% and 13%, respectively. The mean index of discrimination (RRA/RIA) was 1.02, expressed by the slope of the regression curve. The coefficient of correlation was 0.97. In all women, the LH surge was detected by RRA, and the subsequent ovulation was verified within 30 h by endoscopic examination of the ovaries, as well as serum progesterone concentrations of more than 5 ng/ml on the fifth day after ovulation. As shown, prospective ovulation timing can be done by this simple and accurate method. The RRA can be useful in infertility therapy such as artificial insemination.

Adult↗
PubMed1979

Plasma levels of adrenocorticotropin and cortisol in women receiving oral contraceptive steroid treatment.

The secretion rate and plasma concentration of the adrenocortical steroid cortisol is modified in subjects treated with estrogenic and/or progestational steroids. The effects of contraceptive steroids on the secretion of ACTH are poorly documented, however, In the current investigation, we found that concentrations of ACTH and cortisol in plasma obtained at 0800--0900 h from a group of women with normal cyclic menses (n = 4) ranged from 78--120 pg/ml and 77--137 ng/ml, respectively. Although significant cyclic changes in the plasma levels of LH, FSH, 17 beta-estradiol, and progesterone occurred during the ovarian cycle, no obvious cyclic fluctuations in plasma levels of ACTH or cortisol were observed. In women treated with Norinyl 1 + 80 (1.0 mg norethindrone plus 0.08 mg mestranol), plasma concentrations of LH, FSH, 17 beta-estradiol, and progesterone were significantly lower (P less than 0.001) than plasma levels of these hormones in normal women during the ovarian cycle. The mean daily plasma concentrations of ACTH were significantly lower (P less than 0.001), whereas plasma cortisol levels were significantly higher (P less than 0.001) in women treated with oral contraceptive steroids compared to the levels of these hormones in the untreated ovulatory women.

Adrenocorticotropic Hormone↗
PubMed1983

Male sexual development in the monkey. I. Cross-sectional analysis of pulsatile hypothalamic-pituitary-testicular function.

Pulsatile secretion of serum gonadotropins and testosterone was studied in 46 monkeys of varying ages from 9 days of age through adult life. Although some of the hormonal analysis was longitudinal in nature, most comparisons were cross-sectional. On the basis of pulsatile secretory patterns, hCG and GnRH stimulation, skeletal age, testicular volume, and histology, we have arbitrarily defined four developmental age groups: postnatal (less than 7 months), prepubertal or juvenile (7-27 months), pubertal (28-59 months), and adult (greater than or equal to 60 months). In accomplishing the pulsatile studies, blood was withdrawn at 15-min intervals over 24 h without anesthesia using a mobile vest and tether assembly to support an indwelling cannula. GnRH and hCG challenge tests were done on one or more occasions on all animals. Plasma samples were analyzed for concentrations of FSH, LH, testosterone, dihydrotestosterone and delta 4-androstenedione by established RIAs and an in vitro bioassay for LH. During the frequent sampling period of 24-h duration for all except postnatal animals, testosterone pulses of large amplitude (up to 8-fold) occurred in postnatal, pubertal, and adult animals. Pulsatile gonadotropin secretion was seen at all ages; however, the highest pulses (up to 15-fold) occurred in prepubertal animals even though this was an infrequent occurrence. Time series analysis techniques were applied for objective statistical characterization of cyclic patterns. Basic rhythms corresponding to 50- to 90-min frequency cycles in gonadotropin secretion were identified. Substantive differences between LH concentrations by bioassay and RIA were seen infrequently. Our findings illustrate that: 1) circulating gonadotropin and testosterone pulses change in amplitude but not necessarily frequency during pubertal development, and 2) primate models are a useful paradym for the longitudinal study of human male sexual development. We conclude that where direct human investigation may be limited, much can be learned by study of these primate surrogates.

Age Determination by Skeleton↗
PubMed1980

Pharmacokinetic studies on low dose estradiol 17 beta administered orally to postmenopausal women.

Peripheral plasma from four postmenopausal women was analysed for estrone, estradiol, lutenizing hormone and follicle stimulating hormone during 24 hours following an oral intake of a single dose of 1.0 mg micronized estradiol, on the first day of therapy and after one month. A similar study was carried out with another four postmenopausal patients, who received 0.2 mg estradiol three times daily. The measurements were performed by radioimmunoassay (RIA). It is concluded that the plasma concentrations of estrone and estradiol are higher and those of FSH lower after one month of therapy than on the first treatment day, while plasma LH remains unchanged. Micronized estradiol is rapidly absorbed from the gastrointestinal tract and converted to estrone, and the plasma profiles of estrone during the day are more constant with a divided daily dose than with a single higher dose. The divided daily dose results in an equally good clinical effect even though the total administration of estradiol is lower.

Administration, Oral↗
PubMed2000

[Hormonal profile in men with premature androgenic alopecia].

Androgenetic alopecia especially that with premature onset can be a cause of serious psychic trauma. As far as the treatment with antiandrogens, inhibitors of 5 alpha-reductase or hair transplantation represents a heavy economic burden for the patient, we tried to exclude other hormonal causes or to find a criterion for the apt candidates for the treatment in 15 young men with premature androgenetic alopecia. Hormonal analysis discovered a significantly lower plasma level of sexual binding globulin (SHBG) and FSH and nearly significantly higher concentration of 17 alpha-hydroxyprogesterone. These data resemble those in the hormonal pattern of women suffering from the syndrome of polycystic ovaries (PCOS). The finding of premature balding as the possible male phenotype hormonal equivalent of polycystic ovaries is in concordance with several pedigree studies in kindreds with PCOS, which have suggested a genetic basis of this syndrome in some families in which balding and pilosity was frequent in male members.

17-alpha-Hydroxyprogesterone↗
PubMed1977

Evaluation of danazol as an oral contraceptive.

The effect of Danazol as an oral contraceptive in doses of 50, 100, and 200 mg daily for 6 months was studied in 3 groups of 10 women. Both 50 and 100 mg Danazol daily were well tolerated, but one pregnancy occurred among the women receiving 50 mg daily, and 2 pregnancies occurred in women receiving 100 mg daily. There were no pregnancies in women taking 200 mg Danazol daily; however, the side effects were frequent and 5 of the 10 patients withdrew from the study prior to 6 months of therapy. Six patients in this study were followed intensively with blood hormone analysis, vaginal cytology, and pathologic evaluation, and these findings are detailed.

Acne Vulgaris↗
PubMedDate not supplied

[Comparative characterization of radioimmunologic and non-isotopic methods of hormonal analysis].

Measurements of 8 hormones (prolactin, FSH, LH, thyrotropic hormone--TTH, estradiol, hydrocortisone, thyroxine, and triiodothyronine--T3) were carried out by the Amerleit system and radioimmunoassay. The hormones were measured in the same blood plasma samples of patients with various endocrine conditions, admitted for examinations to the Endocrinology Center of the Russian Academy of Medical Sciences. Employment of the Amerleit system for hormonal analysis helps obtain the results compatible with the results of RIA for the majority of the measured hormones. The methods for measuring LH, estradiol and T3 are to be improved. The Amerleit system is highly productive and permits getting the results within 3 hrs. A trained laboratory assistant can measure the hormonal levels in 120-160 blood samples within a day. High efficacy of the technology is indispensable in realization of the screening programs. The absence of radioactivity, safety, high productivity, reliability, accuracy and good storage of the kits (up to 6-8 months) are the principal advantages of the Amerleit system over the existent radioimmunologic systems.

Estradiol↗
PubMedDate not supplied

[Therapeutic results with tamoxifen in oligospermia. II. Hormonal analysis and semen parameters].

During a five months lasting treatment with tamoxifen (2 X 10 resp. 2 X 20 mg daily) a significant increase of testosterone, LH, FSH, estradiol, free testosterone and SHBG was found. The prolactin levels diminished. In semen analysis the values of pH and fructose decreased within the normal range. The sperm density increased significantly, but we could not ascertain whether a dosage of 2 X 20 mg/die will provide better therapeutic results. Furthermore the hormonal and seminal investigations described in this paper did not allow to predict those patients who would be good responders on tamoxifen therapy. Tamoxifen seems to be effective in normo-, but also in hypo- and hypergonadotropic patients.

Estradiol↗
PubMed1977

Bovine cystic ovarian disease: plasma hormone concentrations and treatment.

Ovarian function in 91 dairy cows with cystic ovarian disease was assessed by rectal palpation and by plasma hormone analysis before and after treatment. Plasma analysis showed that 84% of the cysts were correctly classified clinically and only these cows are considered further. Luteinised cysts occurred in 59 cows whereas only 18 had non-luteinised cysts. The mean plasma concentrations of luteinising hormone (LH), follicular stimulating hormone (FSH), progesterone, oestradiol and testosterone were not significantly different when compared with values at relevant stages of the oestrous cycle in normal cows. Success of treatment with progesterone, a synthetic prostaglandin, human gonadotrophin (HCG), or gonadotrophin releasing hormone (GnRH) was not dependent upon prior hormone concentrations, except for the prostaglandin which required active luteal tissue. LH and FSH concentrations in cows with luteinised cysts were not significantly different before and after successful treatment with GnRH or progesterone. Normal luteal function was not always established after treatment of non-luteinised cysts with GnRH.

Animals↗
PubMed1981

[Hypophyseal reaction state during oral contraceptiva (author's transl)].

In 5 normocyclic women, firstly taking a conventional oral contraceptive, Neogynon (50 mcg EE + 250 mcg levo-norgestrel) for 6 months, the levels of LH, 17 beta-E2 and progesterone did not rise after changeover to a dose-reduced pill, Microgynon 30 (30 mcg EE + 150 mcg levo-norgestrel). This fact indicates maintained central suppression. Examination of the hypothalamic-hypophyseal axis by the Gn-RH test (50 mcg) with Microgynon 30 showed negative results during the first treatment cycle in 13 out of 18 women. In the 6th treatment cycle only 7 Gn-RH non-reactive women were observed and after stimulation with 100 mcg Gn-RH only 5 women remained with negative Gn-RH tests. Of the 20 women who took conventional oral contraceptives over a period of 6 months to 6 years (7 took Eugynon: 50 mcg EE + 500 mcg D,L-norgestrel, 5 Lyndiol: 75 mcg mestranol + 2.5 mg lynoestrenol, 8 Neogynon, only one from the Neogynon group showed a positive result. On the other hand there was a positive reaction in 4 out of 7 women using the two step dose-reduced preparation Sequilar (11 tablets of 50 mcg EE + 50 mct levo-norgestrel and 10 tablets of 50 mcg EE + 125 mcg levo-norgestrel).

Adult↗
PubMed1982

Ovulation detection in the human.

The importance of predicting human ovulation for either optimizing or avoiding conception has been considered from an endocrine, morphological and clinical view point. Of the biochemical markers in peripheral blood, a knowledge of the LH peak is the most clearly defined, with a two to four fold increase above baseline levels for a relatively short 24-30 hour preovulatory period. Ovulation is considered to occur 28-36 hours after the beginning of the LH rise or 8-20 hours after the LH peak. Daily assessment of the rise in preovular oestrogen reflects Graafian follicle development but the rise is less distinct and spread over 3-4 days with marked day to day fluctuations. LH induces a marked reduction in oestrogen production some 12 hours prior to ovulation and at the same time induces a two to three fold increase in progesterone production above baseline levels. While these changes in themselves are not great enough for day to day discrimination, a knowledge of their reciprocal relationship may be. The preovular rise in FSH is relatively small compared to LH and the radioimmunoassay technique has not generally been refined to be as rapid and reliable. Monitoring the day to day growth of the preovular follicle ultrasonically is both linear and potentially predictable but there is a wide range of its final diameter (17-26 mm) prior to ovulation making prediction inaccurate. With further refinements in ultrasonic resolution, detection of intrafollicular changes of the cumulus oophorus and granulosal cell layer configuration and thickness may give a closer prediction of the time of ovulation. At a clinical level a knowledge of menstrual cycle length in association with body messages which herald ovulation are useful and may forewarn that ovulation in terms of days is approaching. Such markers as preovulation pain, the detection of periovular cervical mucus and the change in physical character and position of the cervix are reliable signs of preovulation for many well motivated and informed women for either promoting or avoiding conception. A knowledge of the basal body temperature is not a prospective guide to ovulation, but once the thermal shift is established in association with loss of periovular mucus symptoms, the fertile period can be considered to have passed. Because we do not have a precise and simple marker of human ovulation, it is necessary that the most suitable marker of pre- or postovulation is chosen for the particular need in a given individual.

Body Temperature↗
PubMed1975

[Plasma concentrations of LH and of sex steroids during the normal menstrual cycle and during contraceptive treatment].

The authors have studied the long-term effects of combined oestrogen-progestogen on the secretion of gonadotrophins and on ovarian function. Estimation of the radio-immune levels of the plasma concentrations of the pituitary luteinizing hormone (LH), of oestradiol (E2), of progesterone (P) and of 20 alpha hydroxyprogesterone (20 alpha OHP) and of testosterone (T) served as a base for this study. A test cycle and 8 cycles in which a combination of 50 micrograms of ethinyl-oestradiol and 0.5 mg of norgestrel were administered were studied in 4 normal volunteer women. A pituitary stimulation test using 50 micrograms of gonadotrophin "releasing factor" (LH-RH) was given during the 7th cycle. Ovarian stimulation using human menopausal gonadotrophins (HMG) was given during the 8th cycle of treatment in 3 of the women and in 3 other subjects. Pituitary secretion of LH and ovarian secretion of E2 and P are partially inhibited. Pituitary response to the injection of LH-RH stays normal but 5 out 6 patients had no response to stimulation by gonadotrophins. These results allow us to conclude that the lowering of production of gonadotrophins during treatment with combination oestrogen and progestagens is responsible for inhibition of ovarian activity, and that there is a delay before the latter respond to stimulation by either endogenous or exogenous gonadotrophins.

Adult↗
PubMedDate not supplied

Laboratory reproducibility of endogenous hormone levels in postmenopausal women.

We conducted several studies to assess the ability of laboratories to reproducibly measure plasma levels of estrone, estradiol, percent free estradiol, sex hormone binding globulin, dehydroepiandrosterone sulfate, androstenedione, testosterone, progesterone, follicle-stimulating hormone, and prolactin in postmenopausal women. We sent four to seven replicate samples of plasma to each of four well-established endocrine laboratories in the United States on one or two separate occasions. All replicate samples were handled identically during processing, storage, and retrieval, and were labeled to preclude their identification by the receiving laboratory. The within-person coefficient of variation, a measure of laboratory error, was consistently low (< 15%) for follicle-stimulating hormone and prolactin. For estrone and estradiol, hormones present at low levels in postmenopausal women, the laboratory error was often large (> 25%) and the ratio of between-person variation to laboratory error was often less than 2.0. Reproducibility of androstenedione, dehydroepiandrosterone sulfate, testosterone, progesterone, sex hormone binding globulin, and percent free estradiol was also variable. Our results emphasize the need for evaluating laboratory performance before sending samples for hormone analysis and for increased collaboration between epidemiologists and laboratory investigators in improving hormone measurements for use in epidemiological research.

Androgens↗