Follicle-stimulating Hormone Assay Research Articles

A metabolomics approach for the evaluation of Ficus benghalensis female in vivo reproductive effects relative to its metabolite fingerprint as determined via UPLC-MS and GC-MS

Ethnopharmacological relevanceFicus benghalensis, commonly known as Banyan Fig, is the national tree of India and its aerial roots are used traditionally to treat female reproductive disorders. However, despite this traditional use, no pharmacological evidence could be traced supporting this use. Additionally, no comprehensive metabolite profiling was reported for F. benghalensis aerial roots. Aim of the studyThis study attempts to justify biochemically the traditional use of F. benghalensis aerial roots in treatment of female reproductive disorders and in relation to its secondary metabolite profile. Materials and methodsTotal ethanol extract (TEE) and subfractions [petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and n-butanol (BUF] were prepared from air-dried powdered aerial roots of F. benghalensis. Detailed in-vivo investigation of the hormonal activity and action mechanism of the total ethanol extract and subfractions was carried out through evaluation of estrogenic and gonadotropic activities. The estrogenic activity was evaluated on ovariectomized immature female rats through estimating uterine weight, vaginal cornification and serum estradiol level along with histological examination of uteri. The gonadotropic activity was measured by assay of follicle stimulating hormone (FSH) and luteinizing hormone (LH) like activities. Total follicular and corpora lutea counts in immature female rats were used to determine FSH and LH like activities, respectively in addition to histological picture of the genitalia. Comprehensive non-targeted metabolite profiling was carried out for the TEE and subfractions using UPLC-HRMS in negative and positive ionization modes. UPLC-MS fingerprint was subjected to principal component analysis (PCA) and partial least squares analyses to correlate the bioactivities to specific chemical constituents in F. benghalensis different subfractions. GC-MS was further used for non-polar silylated fractions. ResultsResults revealed that only the non-polar PEF and CHF displayed moderate estrogenic and FSH-like activities but with no LH-like activity. Metabolites profiling via (UPLC-HRMS) and multivariate PCA analysis enabled identification and comparison of various chemical classes in F. benghalensis extract and fractions. The active non-polar fractions revealed nearly similar metabolites profile being composed of isoflavonoids, triterpenes, sterols, fatty acids and cyclic peptides. In contrast, polar fractions were more abundant in apocarotenoids, fatty acyl amides, hydroxybenzoates and hydroxycinnamates in addition to two lignans. PLS analysis revealed strong correlation between hydroxylated fatty acids and pyranoisoflavones with estrogenic and FSH-like activities. GC-MS analysis was further employed for non-polar fractions profiling revealing for their enrichment in fatty acids/esters, terpenes, organic acids and phenolics. ConclusionThis is the first study to rationalize the use of F. benghalensis aerial root traditionally in treatment of gynecological disorders, revealing that the petroleum ether and chloroform non-polar subfractions of F. benghalensis showed estrogenic and FSH-like activity with absence of LH-like activity. This biological activity could possibly be attributed to its metabolites profile of isoflavonoids, fatty acids, triterpenes, sterols and cyclic peptides identified via UPLC-MS and GC-MS techniques. Consequently, F. benghalensis aerial roots should be used with caution in traditional treatment of female infertility or other reproductive disorders.

B-040 Performance Evaluation of Five Reproductive Endocrinology Assays on the Atellica CI 1900 Analyzer

Abstract Background The Atellica® CI 1900 Analyzer is an automated, mid-throughput, integrated chemistry and immunoassay analyzer utilizing both Atellica CH and Atellica IM assays. This study was designed to evaluate the analytical performance of the Atellica IM Luteinizing Hormone (LH), Progesterone (PRGE), Follicle Stimulating Hormone (FSH), Prolactin (PRL), and Total hCG (ThCG) assays* on the Atellica CI 1900 Analyzer. Methods The Atellica CI 1900 LH, PRGE, FSH, PRL, and ThCG assays use the same reagents and calibrators as the comparable Atellica IM assays. Precision and method comparison (MC) studies were used as performance indicators for the Atellica CI 1900 Analyzer. Precision studies were performed according to CLSI EP05-A3 using quality control material and native and contrived human serum samples. One aliquot of each sample pool was tested in duplicate in two runs per day ≥2 hours apart on each analyzer for ≥20 days with one reagent lot on two systems. MC studies were performed according to CLSI EP09-A3. Individual native human serum samples were analyzed using the Atellica IM assays on both the Atellica IM and Atellica CI 1900 Analyzers. Results Representative precision and MC results are listed for each assay in the table. Over the five assays tested, repeatability and within-lab %CVs were <7% and <13.5%, respectively. Slopes determined by the Deming linear regression model were approximately equal to 1. Conclusion Evaluation of the Atellica IM LH, PRGE, FSH, PRL, and ThCG assays using the Atellica CI 1900 Analyzer demonstrated good precision and equivalent performance compared to the same assays on the Atellica IM Analyzer. *The products/features mentioned here are not commercially available in all countries. Their future availability cannot be guaranteed.

Effects of sodium valproate monotherapy on reproductive hormonal levels in adult male epileptic patients

Aims: to assess the effects of valproate (VPA) on serum levels of testosterone, prolactin, follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol ( E2) in adult male epileptic patients in comparison to healthy controls. Methods: This study was conducted from Oct. 2005 to Aug. 2006. Patients were received from specialist neurology Clinic. Out of 62 patients interviewed only 46 fulfilled the criteria of selection . Apparently healthy 50 subjects were also included in this study as a control group. Blood samples were taken from both patients and controls and assay of testosterone, prolactin ,FSH, LH and E2 were done using radioimmunoassay (RIA) technique. Body mass index (BMI) was calculated using special equation. Results: There was a significantly higher testosterone levels in epileptic male patients on VPA therapy in comparison to healthy controls, with insignificant effect on the levels of prolactin, FSH, LH and E2. The duration of therapy and the dosage of VPA had no effect on the serum levels of the above measured parameters. Conclusion: VPA monotherapy might be associated with significant increase in serum testosterone level with insignificant effects on serum prolactin, FSH, LH and E2 in adult male epileptic patients and the duration of therapy and daily dose have no influence on such effect.

Prediction of poor ovarian response during In vitro fertilization in Nigerian women: A comparison of basal antral follicle count and follicle-stimulating hormone

Background: Poor ovarian reserve has been shown to be associated with poor outcomes of in-vitro fertilization (IVF) treatment. Women who can be reliably identified as expected poor responders can be advised on chances of poor outcomes that may dissuade them from wasting resources on IVF using their own eggs; and offered donor eggs, especially in a resource-poor country like Nigeria. Many centres routinely perform basal follicle-stimulating hormone (FSH) assay before IVF; however, basal antral follicle count (AFC) has emerged as a more reliable test of ovarian reserve that can be provided at a reduced cost compared to FSH in an IVF clinic setting. The determined predictive values of basal AFC compared to FSH in Nigerian women can be used to predict poor ovarian response during IVF treatment; and also to influence local clinical practice in IVF by offering a more reliable and affordable test, thereby avoiding wastage due to duplicate and unnecessary investigations. Aim: The aim is to determine the diagnostic accuracy of basal AFC compared to basal FSH for the prediction of ovarian response during the IVF cycle in Nigerian women. Patients, Materials and Methods: This was a hospital-based prospective comparative study in two private fertility centres in Abuja. Consecutive 166 women that underwent IVF treatment cycles who met the inclusion criteria were recruited. On day 2 to day 4 of a normal cycle, FSH assay and AFC using the Broekmans’ systematic process were done. They had controlled ovarian hyperstimulation by antagonist or agonist and occasionally long protocols. The poor response was defined as <4 follicles of >17 mm on the day of human chorionic gonadotropin trigger or ≤3 oocytes retrieved. Receiver operating characteristics (ROC) analysis was done to determine the level of the area under the curve (AUC) and optimum cut-off values of FSH and AFC in predicting poor ovarian response. Results: Twenty-eight (16.9%) had poor responses. ROC analysis demonstrated that AFC had the largest (AUC = 0.707, P = 0.001) relative to FSH (AUC = 0.591, P = 0.128). The ROC analysis showed that the optimum cut-off value for the prediction of poor response for AFC was ≤10, which had a higher accuracy of 67.5%, while for FSH was ≥8.15 mIU/ml with a lower accuracy of 61.5%. They both had the same sensitivity of 60.7%; however, AFC had better specificity, negative and positive predictive value, and higher odds ratio for the prediction of poor ovarian response. The positive and negative likelihood ratios of both cut-off values suggest that they may not be useful as diagnostic tests. Conclusion: ROC analysis estimated that AFC more accurately predicts poor ovarian response by its larger and more significant AUC compared to FSH in our population of women.

Onset of normal cycles in postpartum anovulatory dairy cattle treated with kisspeptin.

The efficacy of a long-acting synthetic derivative of kisspeptin (Kp) to initiate normal oestrous cycles was tested in 24 mixed-aged, Holstein-Friesian cows that were 18–25 days postpartum on the day of treatment (D0). Groups of eight cows received saline (Sal) vehicle by intramuscular injection at 8:00 and 16:00 h (Sal-Sal), Kp at 8:00 h and vehicle at 16:00 h (Kp-Sal) or Kp on both occasions (Kp-Kp). The Kp dose was 15 nmol per 60 kg body weight. The ovaries of the cows were examined daily by ultrasonography between D4 and D14. Blood samples were collected from a tail vessel at 0, 2, 4, 8, 10 and 12 h relative to the time of the first injection for luteinizing hormone (LH) and follicle-stimulating hormone assay. Additional samples were collected daily from D4 until D14 and D19, 22, 26 and 29 for progesterone assay. LH surge-like responses were observed in cows treated with Kp at 8:00 h. Ovulation was consistently induced by Kp within 48 h when a dominant ovarian follicle of at least 10 mm in diameter was observed (8/14) but in no cases (6/14) during a new wave of ovarian follicular development comprising follicles <10 mm in diameter. The subsequent ovulatory cycle was of normal length in most cases as compared with short 8– to 12-day cycles observed in spontaneously ovulating cows. We conclude that Kp treatment can induce ovulation in postpartum dairy cows, with ensuing oestrous cycles of normal length, if administered when a mature dominant follicle is present in the ovaries.Lay summaryCow fertility is important for efficient, profitable dairy farming. Cows that take too long after calving to become fertile are problematic. We tested a synthetically made, long-acting hormone called kisspeptin (Kp) to advance the time that cows become fertile after calving. Twenty-four dairy cows that had been calved for 3–4 weeks were used. One group of eight cows received an injection of Kp at the morning milking, another eight cows received Kp at both the morning and afternoon milking, while the last group of eight cows served as untreated controls. Kp treatment caused a desirable hormone response from the cows’ brain. Normal oestrous cycles resulted, but only when a mature follicle was present in the ovary. Further study is required to analyse whether the use of a long-acting Kp drug could be used as an effective treatment for stimulating dairy cows to become more fertile after calving.

Evaluation of potentials of aqueous ethanolic extract of Psidium guajava on reproductive functions of male Albino rats

&#x0D; &#x0D; &#x0D; &#x0D; The therapeutic utilization of plants for managing several diseases by people of all continents, especially Africa, is as old as tradition. The objective of this study was to evaluate the potentials of aqueous ethanolic leaf extract of Psidium guajava on male reproductive parameters. Twenty-four male Albino rats were randomly assigned into four groups of six rats per group. Rats in the control group were administered Tween 20®. Rats in groups 2, 3, and 4 were administered Psidium guajava aqueous ethanolic leaf extract orally at the doses of 100, 200, and 400 mg/kg body weight, respectively, once daily for 60 days. The body weights of the rats were determined at the beginning and end of the experiment. Sperm parameters and some reproductive organs weight of each rat was also determined, some organs were collected for histopathology. Assay for follicle-stimulating hormone, luteinizing hormone, and testosterone was done using Enzyme-Linked Immunosorbent methods. Some reproductive organs were collected for histopathological analyses. The results showed a dose-dependent increase (p&lt;0.05) in the weight of male reproductive organs, sperm parameters, and hormones. The extract significantly increased (P &lt; 0.05) serum testosterone in the group treated 400 mg/kg body weight when treated groups were compared with control. Furthermore, the results of FSH and LH revealed a significant (p &lt; 0.05) increase when the treated groups were compared with the control. The histopathological analysis did not reveal any form of damage to the architectural integrity of the testis. However, there was degenerating germinal epithelium in the group administered 400 mg/kg of the aqueous ethanolic leaf extract of P. guajava. Conclusion: aqueous ethanolic extract of P. guajava has positive effects on male reproductive parameters.&#x0D; &#x0D; &#x0D; &#x0D;

SUN-239 A Case with Elevated Serum Follicle-Stimulating-Hormone (FSH) Concentrations Due to Macro-FSH

[Case report] A 64-year-old woman visited our hospital with the complaint of general malaise and anxiety lasting for two years. She had been diagnosed with idiopathic hypoparathyroidism because of hypocalcemia, hyperphosphatemia, low levels of intact PTH and calcification in the basal ganglia and was taking vitamin D and calcium phosphate. Endocrine workup disclosed high serum FSH [713mIU/mL (Elecsys), reference range for postmenopausal women: 10.5 - 142.8mIU/mL]. Other FSH assay platforms also showed high serum FSH levels: 523mIU/mL for Centaur and 457mIU/mL for Architect. Serum LH and E2 concentrations were 24.8mIU/mL and <11.8 pg/mL, respectively. Human anti-mouse antibody (HAMA) was not detected in the serum. [Methods] Large molecular weight FSH was precipitated using 12.5% polyethylene glycol (PEG) and the precipitation ratio (serum FSH - free FSH) / serum FSH was determined. The molecular size of FSH and IgG-bound FSH were examined using gel filtration chromatography (GFC) and protein G column (PG), respectively. Binding study was performed by incubating 125I-FSH (24000 cpm) with the serum and the specificity was examined by adding unlabeled FSH, LH, α-subunit of hCG, β-subunit of hCG and TSH. FSH was measured by enzyme immunoassay (EIA). We also examined these parameters in sera from 10 females with serum FSH>70 mIU/mL as controls. [Results] The dilution curve of the patient’s serum FSH was in parallel with that of reference one. The precipitation ratio by PEG was significantly elevated (95.4% vs. 20.3 ± 8.0% in controls). On GFC, a large proportion of serum FSH (89.4%) was eluted in the position of molecular weight > 150kDa, whereas FSH was mostly eluted at 36kDa in controls. Incubation of reference FSH (300mIU and 600mIU) with the serum (100µL) increased the peak levels of the large molecular weight FSH to be 84.0mIU/mL and 180.0 mIU/mL, respectively, from 21.6mIU/mL without exogenous FSH. Serum FSH bound to PG at a significantly higher ratio (87.3%) than that in controls (1.2 ± 1.4%). A significantly higher amount of 125I-FSH bound to the serum (5691 cpm) than that in controls (704 ± 170 cpm). The binding was displaced with an excess amount (600IU/mL) of unlabeled FSH to be 821 cpm (14.4%) and Scatchard analysis revealed the presence of anti-FSH autoantibodies (Ka 3.0 x 107 M-1, Bmax 0.92 x 10-8 M). The binding was slightly displaced by other hormones that shared α-subunit; 87.6% by LH, 90.4% by hCGα, 81.1% by hCGβ and 71.8% by TSH. [Conclusion] We report a woman with high serum FSH concentrations due to macro-FSH caused by anti-FSH autoantibodies. The possibility of macro-FSH should be considered in a case with unexpectedly high serum FSH concentrations.

A study of Inhibin a-16C&gt;T gene polymorphism and other reproductive hormones in the diagnosis of Premature Ovarian Failure in Egyptian Women undergoing ICSI

Background: Premature ovarian failure (POF) or poor ovarian response is a syndrome characterized by amenorrhea for duration of 4-6 months or more, hypo-estrogenism and elevated serum FSH concentration (FSH > 40 IU/1) before age of 40 years. The POF may arise through different events including: reduction of primordial follicle pool; acceleration of follicular atresia; alteration of primordial follicles maturation/recruitment Inhibin is a product of granulosa cells. Onset of POF can be a result of inhibin alpha genetic abnormalities, the most extensively studied genetic abnormality is 16C>T polymorphism. Objectives: The present study was undertaken to evaluate whether INHa-16C>T gene polymorphism could be a predictive marker for POF in the Egyptian females undergoing ICSI. Also, to identify role of the ovarian hormones like; FSH, LH, estradaiol, progesterone, anti-mullarian hormone, activin and inhibin in the dignosis of POF. Patients and methods: All cases were recurted from the Gynecology and Obstetric Department, Mansoura University, Egypt. All women were subjected to history taking including demographic data, menstrual and reproductive characteristics and history of hormonal therapy. The study group included 50 women with symptoms and signs suggestive of ovarian failure; their mean age was 31.5± 4.88 years. The control group included 50 healthy women with normal hormonal profile with male factors infertility, their mean age 29.05+3.3 years. Venous blood samples were collected for assay of follicle stimulating hormone (FSH), luteinizing hormone (LH), anti-mullarian hormone (AMH), inhibin and activin using enzyme linked immunosorbent assay. Genomic DNA was isolated from EDTA-anticoagulated peripheral blood using DNA extraction kit. Results: Women with POF showed a highly significant increase in the mean FSH and LH levels compared to control group. There is a statistically significant decrease of the mean serum inhibin and inhibin/activin ratio in women with POF compared to control women. In women with POF, the frequencies of homozygous genotypes CC, heterozygote genotypes CT and homozygote genotypes TT of the INHa-16C>T polymorphism were 70% (35/50), 24% (12/50) and 6% (3/50). The C allele was present in 82% of patients with POF and in 73% of the control group, whereas the T allele was present in 12% and 27% respectively. Conclusion: INHa-16C>T gene polymorphisms is not considered as a risk factor for POF among Egyptian women.

Haemogram and hormonal profile of WAD bucks treated with leaf ethanolic extract of &lt;i&gt;Spondias mombin&lt;/i&gt;

Twelve post pubertal West African Dwarf (WAD) bucks of average age of 1.8 ± 0.19 years and average weight of 8.76 ± 0.72 kg were experimentally treated orally with 800mg/kgBW of ethanol extract of Spondias mombin. Experiment was carried out in the month of August at Abeokuta, South West Nigeria with temperature range of 24oC and 33oC on Latitude 07o 10’N and Longitude 03o 2’E. The goats had well-formed pendulous scrotum with well descended free bilateral testicles. The goats were kept in a standard goat pen and fed with fresh green centrosema leaves in the morning and commercially prepared ration in the evening. Water was served ad libitum. Pre-treatment haematological and hormonal profiling after aseptical collection of blood from the bucks was followed with 14 days of oral treatment with ethanol extract of Spondias mombin at the dosage rate of 800mg/kg BW. Post-treatment haematological and hormonal studies followed after. The hormones assayed were follicle stimulating hormone (FSH), luteinising hormone (LH), testosterone, progesterone, estrogen and prolactin. The haemogram revealed significantly (p &lt; 0.05) higher PCV (24.33±0.6 %) in pre-treatment compared to 19.92± 1.1 % recorded for post-treatment. However, both were lower in values than normal range. WBC and RBC did not change significantly but while RBC values for pre-treatment and post-treatment fell within normal range, only the WBC value of post-treatment did. Assay of FSH, progesterone, estrogen and prolactin revealed no significant (p . 0.05) difference between pre-treatment and post-treatment; however, noticeable higher pre-treatment LH concentration and elevated post-treatment testosterone concentration were observed. The work showed that Spondias mombin at 800mg/kgBW has profertility tendencies in the buck and can be considered as an aphrodisiac in goat breeding in developing countries probably at a higher dosage.Keywords: Buck, Ethanol, Extract, Leaf, Spondias mombin

Effect of omeprazole on reproductive hormonal levels and sexual function in male patients with peptic ulcer disease

Objectives: To assess the effects of Omeprazole at a fixed dose on sexual function (by self reporting questionnaire) and serum levels of testosterone, prolactin, follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in newly diagnosed male patients with peptic ulcer disease, in comparison to healthy controls. Patients and Methods: Patients were received and interviewed in Ibn- Sina Hospital, out-patient clinic, medical section and were selected according to certain criteria. From 60 male patients selected only 43 completed the follow-up study. Initially sexual function were assessed using a self-reporting questionnaire and a blood samples were taken with assay of serum testosterone, prolactin, FSH, LH and E2 using Minividus technique. Patients than put on Omeprazole therapy 20mg twice daily for 45 days with Clarithromycin 500mg twice daily and Metronidazole 500mg twice daily for the initial 10 days. By the end of the suggested period of follow-up, sexual function were assessed by self-reporting questionnaire and other blood samples were taken with assay of serum testosterone, prolactin, FSH, LH and E2 using the same analytical technique. Included in this study also forty apparently healthy age, matched male volunteers as a control group . Blood samples were also withdrawn from them and assay of serum testosterone prolactin, FSH, LH and E2 were done , with assessment of sexual function using the same self reporting questionnaire. For both groups ( the patients and controls), body mass index were calculated using special equation. Results:There was insignificant differences in the parameters of sexual function between patients with peptic ulcer disease in the pre-therapy stages and the controls, so as in the serum levels of testosterone, prolactin, FSH, LH and E2. After therapy there was a significant differences in the mean serum level of testosterone from the pre-therapy stage and the controls, with insignificant differences in the sexual function parameters and serum levels of prolactin, FSH, LH and E2.Conclusion: Long term Omeprazole therapy might cause a reduction in the level of testosterone hormone with insignificant effects on other sexual hormonal levels (prolactin, FSH, LH and E2) within significanteffect on sexual function, in male patients with peptic ulcer disease.

Serum hormonal levels in HIV/AIDS infected male subjects on antiretroviral therapy (ART) in Nnewi, Nigeria

This study was designed to assess the serum hormonal levels (Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), Testosterone, Estrogen, Progesterone, Prolactin and Cortisol) in symptomatic HIV/AIDS male subjects on ART. A total of 411 participants aged between 17 and 58 (43 ±10) years were randomly recruited for the study. The participants were staged and grouped as follows: symptomatic HIV/AIDS male subjects on ART (n= 139), symptomatic HIV/AIDS male subjects not on ART (n= 136) and HIV seronegative subjects (n= 136). Blood samples were collected from the participants for the determination of HIV status by immunochromatography and HIV confirmation by Western Blot. Enzyme Linked Immunosorbent assay (ELISA) was used to assay for FSH, LH, Testosterone, Estrogen, Progesterone, Prolactin and Cortisol levels. Their results showed significant rise in FSH, LH in HIV seropositive participants on ART compared respectively to those not on ART and the HIV seropositive subjects (p< 0.05). On the other hand, the Estrogen, Progesterone, Prolactin and Cortisol were all raised in symptomatic HIV seropositive participants not on ART compared respectively to the symptomatic HIV seroposive on ART and the HIV seronegative groups (p< 0.05). The testosterone levels significantly decreased in both symptomatic HIV infected subjects on ART and those not on ART compared with the HIV seronegative controls (p< 0.05). The relevance of HIV infection on the serum hormonal status is that HIV infection causes hypogonadism as well as primary testicular failure in symptomatic HIV participants not on ART. Also, HIV infection causes hyperprolactinaemia, hyperprogesteronaemia and hypercortisolism in symptomatic HIV participants not on ART.

Development of a homologous enzyme-linked immunosorbent assay for European sea bass FSH. Reproductive cycle plasma levels in both sexes and in yearling precocious and non-precocious males

Since the late 1980s, gonadotropins have been isolated and characterized in several fish species, but specific immunoassays for the follicle-stimulating hormone (FSH) have only been developed for a few. The present study reports the development and use of a specific and homologous competitive ELISA for measuring FSH in European sea bass (Dicentrarchus labrax) using a recombinant FSH and its specific antiserum. Recombinant European sea bass FSHβ and FSH heterodimer were produced in the methylotrophic yeast Pichia pastoris and a baculovirus expression system, respectively. Specific polyclonal antibodies, generated by rabbit immunization against recombinant FSHβ, were used at a final dilution of 1:8000. Recombinant FSH heterodimer was used to generate a standard curve and for coating of microplates (166μg/ml). The sensitivity of the assay was 0.5ng/ml [B0-2SD], and the intra- and inter-assay coefficients of variation were 2.12% (n=10) and 5.44% (n=16) (Bi/B0 ∼45%), respectively. A high degree of parallelism was observed between the standard curve and serially diluted plasma and pituitary samples of European sea bass.The ELISA developed was used to study the plasma FSH profiles of mature males and females during the reproductive cycle, and those of immature juvenile males under different light regimes. The analysis showed that FSH increased significantly during the intermediate stages of spermatogenesis and during vitellogenesis. Analyses in immature juvenile males showed that the continuous light photoperiod significantly reduced plasma FSH levels, and consequently, testicular growth and precocious puberty. In conclusion, the immunoassay developed has proven to be sensitive, specific and accurate for measuring European sea bass FSH, and it represents a valuable tool for future studies on the reproductive endocrinology of this species.

Effects of quetiapine on body mass index and reproductive hormonal levels in male schizophrenic patients

Objective: To asses the effects of quetiapine therapy on body mass index (BMI) and serum level of testosterone, prolactin, follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in newly diagnosed male schizophrenic patients in comparison to healthy controls. Patients and methods: Thirty male patients with schizophrenia were included in this study. The diagnosis of schizophrenia was made according to DSM-IV criteria of the American Psychiatric Association (APA). Another thirty apparently healthy male individuals were included in the study as a control group. Blood samples were taken initially from patients and controls and assay of serum testosterone, prolactin, FSH, LH, and E2 were done using enzyme –linked fluorescent immunoassay (ELFA) technique by vidas instrument. Later, after 6 months of quetiapine therapy, other blood samples were taken and assay of the same parameters were done. Calculation of BMI was done for the patients and controls using special equation. Results: There was an insignificant difference in the mean BMI, serum testosterone, prolactin, FSH, LH, and E2 levels between patients in the pre-therapy stage group and in both the post-therapy stage group and controls. Conclusion: Chronic quetiapine therapy might be regarded as a safe drug with regard to effects on BMI and serum levels of reproductive hormones in newly diagnosed male schizophrenic patients. Keywords: Schizophrenia, quetiapine, BMI, testosterone, prolactin, FSH, LH, E2.

Redefining abnormal follicle‐stimulating hormone in the male infertility population

Study Type - Diagnostic (validating cohort). Level of Evidence 2a. What's known on the subject? and What does the study add? FSH is a hormone released by the anterior pituitary gland via stimulation from gonadotrophin-releasing hormone and potentially other factors. FSH reflects the status of spermatogenesis (i.e. the ability to produce sperm) as a result of the feedback between the testis and hypothalamus/pituitary glands. An elevated FSH level is indicative of abnormal spermatogenesis and may indicate primary testicular failure. The range for 'normal' FSH varies somewhat between institutions but has been defined by the Strong Memorial Hospital (Rochester, NY, USA) clinical laboratory as 1.4-18.1 IU/L based on the ADVIA Centaur (Siemens Medical Solutions, Tarrytown, NY, USA) FSH assay. The findings obtained in the present study could be helpful for predicting male factor infertility in patients with a borderline high FSH level (≈4.5 IU/L) and a low testosterone level compared to someone with a borderline high FSH level and a normal testosterone level. Although the 'normal' range for FSH is qualified as a value in the range 1.4-18.1 IU/L, the present study shows that an FSH level >4.5 IU/L was associated with abnormal semen analysis in terms of morphology and sperm concentration in the present patient population. Therefore, these findings suggest that FSH values lower than those currently considered normal may be associated with abnormal semen analysis, and that the 'normal' range for FSH used in clinical settings may need to be reconsidered. • To examine the correlation between follicle-stimulating hormone (FSH) and testosterone/FSH levels with semen analysis parameters to evaluate whether the range for judging normal FSH levels should be reconsidered. • The present study included 610 male infertility patients from a single urology infertility clinic between 2004 and 2008. • Patients (n=153) were excluded for obstructive azoospermia, hypogonadotrophic hypogonadism, steroid use or failure to complete testing. • Abnormal semen analysis values were based on the WHO 1999 criteria. • We performed t-tests, anova, chi-squared tests and logistic regression to statistically examine the association between the FSH (or testosterone/FSH ratio) level and semen parameters. • The FSH level showed statistically significant associations, as well as evidence of a dose response, with abnormal sperm concentration and morphology but not with semen volume. • In men with FSH levels >7.5 IU/L, the risk of abnormal semen quality was five- to thirteen-fold higher than that of men with FSH levels <2.8 IU/L depending on the specific semen parameter. • Similarly, semen parameters were had a greater probability of being abnormal with decreasing testosterone/FSH ratios. • A significantly increased risk of abnormal semen analyses among men with FSH levels >4.5 IU/L and decreasing testosterone/FSH ratios suggests that redefining normal FSH in infertile men would be valuable.

Action of aromatase inhibitor for treatment of uterine leiomyoma in perimenopausal patients

To assess the effect of the aromatase inhibitor on patients with leiomyoma in the reproductive stage regarding reduction of uterine volume and control of symptoms. Clinical study. Academic clinical practice. Twenty patients, over 35 years of age, with symptomatic uterine leiomyoma. Anastrozol, 1 mg/day for 12 weeks. Measurement of uterine volume, assessment of symptoms related to uterine leiomyoma, serum assay of follicle stimulating hormone (FSH), and estradiol. Average reduction of uterine volume of 9.32%, attaining up to 32%, and reduction of symptoms of uterine leiomyoma (menstrual volume, duration of menstruation, and dysmenorrhea). No significant change in serum levels of FSH and estradiol during use of the medication were observed. Anastrozol proved to be effective in reducing the volume of the uterus-leiomyoma structure, leading to the control of symptoms connected with the disorder without changes in serum FSH and estradiol.

Dry eye in post-menopausal women using hormone replacement therapy

Purpose To evaluate the effect of hormone replacement therapy (HRT) on dry eye in post-menopausal women. Methods Forty post-menopausal women with dry eye (20 patients, group 1) and without dry eye (20 patients, group 2), and planning to receive HRT (estrogen plus progesterone), were recruited as the study groups. Forty age-matched untreated women were enrolled as controls (group 3 with dry eye, 5 patients; group 4 without dry eye, 35 patients). Patients having at least one of the symptoms (dryness, itching, photophobia, foreign body sensation, and tearing) together with two of the tests with positive results for dry eye (tear film break-up time (BUT), fluorescein staining of the cornea, analysis of the meibomian gland, and Schirmer I test) in both eyes were considered dry eye positive. Hormonal assay for follicle stimulating hormone, luteinizing hormone, estradiol, and free testosterone was performed. Dry eye statuses in the groups were evaluated statistically. Results Four patients with incomplete follow-up data were excluded. HRT use increased estrodiol levels in the groups. Mean ages of patients (50.2 ± 4.8 and 50.7 ± 3.9 years, and 50.0 ± 4.6 and 53.0 ± 3.9 years) were similar ( p = 0.67). Duration of menopause in groups 1 and 2 (3.2 ± 2.2 and 1.4 ± 1.2 years; p = 0.01), and in groups 3 and 4 (3.0 ± 1.6 and 1.7 ± 1.3 years; p = 0.014) were different. At the third month examinations, all of the patients in group 1, and 11 patients (61.1%) in group 2 had dry eye ( p = 0.003). Conclusion Duration of menopause and use of HRT may increase the incidence of dry eye in post-menopausal woman.

P-718: Effects of low ovarian reserve versus maternal age on ovarian responsiveness, preembryo quality and clinical outcome in assisted reproduction

Decreased ovarian reserve is associated with a decline in oocyte number and quality, higher Day 3 (D3) serum Follicle Stimulating Hormone (FSH) levels, an increase in chromosome abnormalities and potentially lower pregnancy rates. However, it is not clear whether the association between decreased ovarian reserve and preembryo quality is causal. In addition, the relative importance of chronological versus biological age in declining fertility rates is controversial. The objectives of the present study were to determine 1) the effect of basal D3 FSH (a marker of ovarian reserve) on preembryo quality in ART patients, and 2) the relative importance of maternal age and D3 FSH on declining pregnancy rates. A retrospective analysis of 1381 ART patients attending the Weill-Cornell Center for Reproductive Medicine and Infertility from 2002 to 2005 was performed. Patients were grouped in two ways: 1) according to female age (< 34 yrs, 34-36 yrs, 37-39 yrs, and ≥40 yrs) and 2) based on their D3 FSH values (<5 mIU/mL, 5-9 mIU/mL, and ≥10 mIU/mL). Outcome variables included the number of oocytes harvested, fertilization rate, preembryo grade, clinical pregnancy rate (CP), ongoing/delivered pregnancy rate (OP), and implantation rate (IR). The percentage of mature eggs was determined in a subset of patients who underwent ICSI, using the same subgroups as above. Serum FSH was measured on Day 2 or Day 3 of a patient’s cycle prior to their IVF treatment using the Immulite FSH assay (Diagnostic Products Corporation, Los Angeles, CA). Oocyte harvest occurred 35 hr post-hCG administration and fertilization was assessed 15-18 hr post-insemination or ICSI. Embryo transfer was performed on Day 3 post-harvest. Preembryos were selected for transfer based on morphology and rate of development. An Embryo Grade was assigned to the best quality preembryo being transferred based on these criteria [on a scale of 1 (highest quality) to 5 (extremely poor quality)]. Logistic regression and two-way analysis of variance were used for the analysis of binary and continuous outcomes, respectively. High D3 FSH levels and advanced maternal age both significantly reduced the number of oocytes retrieved per patient, but no effects were observed on either the percentage of mature eggs retrieved or the rate of fertilization. There was a significant difference in the distribution of preembryo grades between the various age groups (p=0.003), arising primarily from a shift of Grade 1 prembryos to Grade 2 prembryos in older patients. In contrast, no significant differences in grade distribution between FSH patient groups were observed. Maternal age was strongly associated with implantation and pregnancy outcome. Both CP and OP were significantly reduced in patients 37 years of age and older. There was a marginally significant decline in CP in the middle FSH group (5-9 mIU/ml)as compared to the lowest FSH group. However, no effect of D3 FSH on OP or IR was seen. Interactions between age and FSH in the study population were not significant. The successful outcome of a patient’s cycle was much more dependent on maternal age than ovarian status, at least when using D3 FSH as a marker of ovarian status in an IVF Program where higher FSH values result in cycle cancellations. Based on these data, maternal age should be considered more strongly than D3 FSH when counseling patients regarding their chances of achieving pregnancy.

Enzyme immunoassay method for total urinary follicle-stimulating hormone (FSH) beta subunit and its application for measurement of total urinary FSH

Objective: To develop and validate an enzyme-linked immunosorbent assay (ELISA) for total urinary follicle-stimulating hormone (FSH) that can be used to evaluate frozen-thawed urine samples. Design: A method was developed for dissociating urinary FSH into subunits by boiling. An ELISA for the free beta subunit of FSH was used to measure total FSH in urine samples collected from male and female volunteers. The urinary FSH values were compared with serum FSH concentrations in paired samples. Setting: A clinical research laboratory. Patient(s): Serum and urine samples from normal male and female volunteers and from men with infertility were used to develop and validate an assay for total urinary FSH. Intervention(s): None. Result(s): Urinary FSH concentrations in frozen-thawed urine were highly correlated to matched serum FSH concentrations. Conclusion(s): A microtiter plate ELISA for FSH beta subunit provides an economical method for evaluating FSH secretion in men and women. This method has practical applications for population-based epidemiologic studies in which urine samples are collected in the held and stored frozen before analysis.

Importance of Selection of Separation System in the Development of Enzyme Immunoassay: An Experience with Follicle Stimulating Hormone (FSH) Assay

An antiserum to follicle stimulating hormone (FSH) obtained as a gift from National Institute of Health (NIH), U.S.A. could not be adsorbed on microtitre ELISA plates, although two other FSH antisera raised in authors' laboratory could be adsorbed. A good precision profile for the FSH assay using these three antisera could be achieved with only one separation system viz. solid phase anti rabbit gamma globulin (ARGG), out of the five separation systems tried. The study suggests that a few antisera used for radio-immunoassay (RIA) purposes may not by themselves get adsorbed on plastic plates. However, they could be effectively used for ELISA purposes using solid phase second antibody.

Elephant pituitary gonadotropins

We describe for the first time the purification and some properties of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) isolated from anterior pituitary tissue of the African elephant ( Loxodonta africana). Methodology previously applied to equine and donkey pituitaries was used to obtain purified preparations of elephant LH and FSH in yields of 8.8 and 0.48 mg, respectively, per 10 g pituitary powder. The preparations were characterized by HPLC gel filtration and amino acid analysis, both of which showed the elephant LH and FSH to be very similar to ovine LH and FSH. The preparations were also characterized by radioimmunoassays and bioassays for LH and FSH and a radioreceptor assay for FSH. Results showed virtually no cross-contamination of hormonal activities in the elephant LH and FSH preparations. Elephant LH potencies ranged from 50 to 66% of highly purified ovine LH and elephant FSH potencies ranged from 21 to 52% of highly purified ovine FSH in the various assays employed. No evidence was found for any demonstrable intrinsic FSH activity in elephant LH. The assays employed suggest possible usage for making physiological measurements of gonadotropins in the elephant.