Inhibin concentrations in mares with granulosa cell tumors

The hormone-producing equine granulosa cell tumor (GCT) may secrete high levels of inhibin. Measurement of inhibin concentrations may be useful in the diagnosis and conformation of mares with GCT. Inhibin may be measured using RIA, which recognizes dimeric alphabetaA-inhibin as well as the monomeric (free) inhibin alpha-subunit, or using a two-site immunoradiometric assay (IRMA) specific for alphabetaA-inhibin. The objective of this study was to examine concurrent relationships among alpha-inhibin (as measured using RIA), alphabetaA-inhibin (as measured using IRMA), and other hormones (testosterone, estradiol, LH, FSH) in mares with GCT. Hormone concentrations were measured in single serum or plasma samples obtained from 22 mares with GCT and from 31 normal cycling mares. One GCT mare had blood samples collected at 12-h intervals for 21 days, and at 15-min intervals for two 6-h periods during that time. Results showed that in GCT mares alpha-inhibin was increased to a greater extent, was more uniformly elevated, and had a less variable secretory pattern than did alphabetaA-inhibin. Concentrations of alpha-inhibin and tumor mass were positively correlated (P < 0.01). Concentrations of LH were higher (P < 0.02) in GCT mares than control mares and were positively associated with testosterone concentrations (P = 0.05). Concentrations of FSH tended to be lower in GCT than control mares and were inversely related with alphabetaA-inhibin in GCT mares. Testosterone and estradiol concentrations were variable. It was concluded that immunoreactive alpha-inhibin reflected detection of both alphabetaA-inhibin and free a-subunit. Free alpha-subunit was evidently secreted at a relatively steady rate dependent upon mass of the GCT, whereas secretion of alphabetaA-inhibin was more responsive to FSH regulation. Determination of alpha-inhibin using RIA appeared to be a more reliable indicator of the presence of a GCT than specific measurement of alphabetaA-inhibin using IRMA.     

Validation of a high throughput method for serum/plasma testosterone using liquid chromatography tandem mass spectrometry (LC-MS/MS)

Testosterone, the major androgenic hormone in humans, is commonly measured to aid in the diagnosis of clinical conditions related to its excess or deficiency. In addition, testosterone measurements are used to monitor testosterone replacement-, or antiandrogen therapy. Most commonly, automated direct immunoassays have been used to measure testosterone in human serum. Their advantage compared with other methodologies, lies in high- and rapid sample throughput with minimal human intervention. However, many automated testosterone immunoassays suffer from poor accuracy at the low concentration levels (<50ng/dL) seen in women and children, or in men undergoing anti-androgen therapy. Our objective was to develop a LC-MS/MS method which measures testosterone in human serum while fulfilling the following criteria: Rapid pre-analytical sample processing with minimal manual sample manipulation; Minimize sample volume requirements; Accurate, precise and unambiguous measurement; Functional sensitivity of 5-10ng/dL; Sample throughput of at least 30 samples per hour. Our validation criteria for precision, accuracy, and linearity was to have accuracy and linearity within mean limits of +/-10%; Intra and inter-assay precision of <15% throughout the reporting range. We also wanted to compare our results to a previously validated LC-MS/MS assay which utilized a manual liquid-liquid extraction and to an automated commercial immunoassay (Bayer ACS:180). We describe here a sensitive and rapid testosterone assay based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) utilizing on-line sample extraction and multiplexing.     

A prospective assessment of androgen levels in patients with autistic spectrum disorders: biochemical underpinnings and suggested therapies

Impairments in social relatedness and communication, repetitive behaviors, abnormal movement patterns, and sensory dysfunction characterize autism spectrum disorders (ASDs). Seventy consecutive patients with an ASD diagnosis (DSM-IV criteria, >/= 6 years-old) who presented to the Genetic Centers of America for outpatient genetic/developmental evaluations from 2005-2007 were examined. Patients were evaluated using CLIA-approved Laboratory Cooperation of America (LabCorp) testing for: serum testosterone, serum free testosterone, % free testosterone, serum/plasma dehydroepiandrosterone (DHEA), androstendione, and follicle-stimulating hormone (FSH). Morning blood samples collected following an overnight fast, compared to the pertinent reference means, showed significantly increased relative mean levels for: serum testosterone (158%), serum free testosterone (214%), percent free testosterone (121%), DHEA (192%), and androstenedione (173%). By contrast, compared to the pertinent reference mean, the relative mean level of FSH (51%) was significantly decreased. Additionally, at least one of the androgen attributes examined exceeded its recognized laboratory age- and sex-specific reference range in 81.4% (57 of 70) of the patients examined. With respect to their age- and sex-specific reference ranges, females had significantly higher overall mean relative testosterone and relative free testosterone levels than males. Increased androgens in patients diagnosed with ASDs may involve cyclical interactions between the androgen and the transsulfuration pathways, particularly following mercury exposure. A review of therapies that have significantly improved clinical outcomes in ASD patients indicates they share commonality in helping lower androgens. Thus, androgens should be routinely clinically measured in patients with an ASD diagnosis and appropriate androgen-lowering therapies considered for those who have significantly elevated levels.     

Hormonal adaptations and modelled responses in elite weightlifters during 6 weeks of training

The concentrations of serum testosterone, sex-hormone-binding-globulin (SHBG) and luteinizing hormone (LH) were examined throughout 1-year of training in six elite weightlifters. A systems model, providing an estimation of fatigue and fitness, was applied to records of training volume and performance levels in clean and jerk. The analysis focused on a 6-week training period during which blood samples were taken at 2-week intervals. A 4-week period of intensive training (period I) could be distinguished from the following 2-week period of reduced training (period II). During period I, decreases in serum testosterone (P less than 0.05) and increases in serum LH concentrations (P less than 0.01) were observed; a significant correlation (r = 0.90, P less than 0.05) was also observed between the changes in serum LH concentration and in estimated fitness. The magnitude of LH response was not related to the change in serum androgens. On the other hand, the change in testosterone:SHBG ratio during period II was significantly correlated (r = 0.97, P less than 0.01) to the LH variations during period I. These finding suggested that the LH response indicated that the decrease in testosterone concentration was not primarily due to a dysfunction of the hypothalamic-pituitary system control, and that the fatigue/fitness status of an athlete could have influenced the LH response to the decreased testosterone concentration. The negative effect of training on hormonal balance could have been amplified by its influence on the hypothalamic-pituitary axis. A decrease in physiological stress would thus have been necessary for the completion of the effect of LH release on androgenic activity.     

Serum testosterone concentration in response to exogenous GnRH does not predict service rates or pregnancy rates by yearling, performance tested, crossbred bulls

Ten bulls with a scrotal circumference of less than 30 cm at the end of growth performance testing, and 10 cohorts of the same age, size and breed type with a scrotal circumference greater than 30 cm were used to evaluate if testosterone response following GnRH administration could be used to test for fertility, for semen quality, and for specific pathologic testicular parenchymal changes. Serum testosterone concentrations were determined immediately before and 2 to 3 hours following intramuscular injection of 250 ug GnRH. Bulls were examined for breeding soundness, then fertility was tested in a breeding trial; testicular histology was assessed by determining the percentage of cross-sections of seminiferous tubules with no spermatocytes. The mean (+/- SEM) post-GnRH serum testosterone concentration for all bulls was 11.71 (+/-0.64) ng/ml. In order to examine for an association, the GnRH response was classified as above or below the mean for resultant serum testosterone concentration. The GnRH response classification was not related to the scrotal circumference, percentage of tubules devoid of spermatocytes, or percentage of progressively motile spermatozoa (P > 0.10). The percentage of morphologically normal spermatozoa was significantly higher (P < 0.05) in the bulls with a higher than mean testosterone secretion in response to GnRH injection. In the breeding trial, the percentage of heifers bred and the percentage of heifers pregnant (60 days post breeding) were not significantly different (P > 0.10) between the 2 classifications of GnRH response. The GnRH response test was related to the percentage of morphologically normal spermatozoa but did not predict fertility of yearling bulls in this study.     

Proteomic Profiling of a Mouse Model for Ovarian Granulosa Cell Tumor Identifies VCP as a Highly Sensitive Serum Tumor Marker in Several Human Cancers

The initial aim of this study was to identify novel serum diagnostic markers for the human ovarian granulosa cell tumor (GCT), a tumor that represents up to 5% of all ovarian cancers. To circumvent the paucity of human tissues available for analyses, we used the Ctnnb1(tm1Mmt/+);Pten(tm1Hwu/tmiHwu);Amhr2(tm3(cre)Bhr/+) transgenic mouse model, which features the constitutive activation of CTNNB1 signaling combined with the loss of Pten in granulosa cells and develops GCTs that mimic aggressive forms of the human disease. Proteomic profiling by mass spectrometry showed that vinculin, enolase 1, several heat shock proteins, and valosin containing protein (VCP) were more abundantly secreted by cultured mouse GCT cells compared to primary cultured GC. Among these proteins, only VCP was present in significantly increased levels in the preoperative serum of GCT cancer patients compared to normal subjects. To determine the specificity of VCP, serum levels were also measured in ovarian carcinoma, non-Hodgkin's lymphoma and breast, colon, pancreatic, lung, and prostate cancer patients. Increased serum VCP levels were observed in the majority of cancer cases, with the exception of patients with lung or prostate cancer. Moreover, serum VCP levels were increased in some GCT, ovarian carcinoma, breast cancer, and colon cancer patients who did not otherwise display increased levels of widely used serum tumor markers for their cancer type (e.g. inhibin A, inhibin B, CA125, CEA, or CA15.3). These results demonstrate the potential use of VCP as highly sensitive serum marker for GCT as well as several other human cancers.     

Diagnosis of a vulvar granular cell tumor by fine needle aspiration biopsy. A case report

BACKGROUND: Granular cell tumor (GCT) (granular cell "myoblastoma") is an uncommon neoplasm that may mimic carcinoma both clinically and morphologically. Fine needle aspiration diagnosis of vulvar GNT has been described on only one prior occasion. CASE: A 74-year-old, black female presented with a mass in the left labia. Fine needle aspiration biopsy revealed rare intact cells; abundant, granular, cytoplasmic fragments; and bland, ovoid, stripped nuclei. The intact cells were arranged in loose aggregates. Each sampling was exquisitely painful to the patient despite the use of local anesthesia. CONCLUSION: Cytologists should be aware of the distinctive clinical and morphologic appearance of GCT. The cytologic findings of vulvar, GCT are identical to those described at other body sites. Definitive diagnosis before extirpation permits definitive therapy.     

Chronic administration of dehydroepiandrosterone (DHEA) to female monkey and rat has no effect on mammary gland histology

Dehydroepiandrosterone (DHEA), the major steroid precursor of androgens and estrogens produced in peripheral tissues in primates, has been shown to exert chemopreventive effect on the development of carcinogen-induced rat mammary tumors. Since little is known on the effect of DHEA administration on mammary gland physiology and histology, we have studied the effect of long-term administration of DHEA to normal female monkey and rat on mammary gland histology as well as on serum DHEA, DHEA sulphate (DHEA-S), testosterone and estradiol levels. In monkeys, DHEA treatment (2 or 10 mg/(kg b.w.day)) induced a dose-related increase in serum DHEA and DHEA-S (above 20-fold) levels. At the highest dose of DHEA, serum testosterone levels were significantly increased (three- to fourfold), while serum estradiol concentration was not modified. DHEA treatment did not modify the histological characteristics of monkey mammary glands. In the rat, following DHEA administration (10 or 100 mg/(kg b.w.day)), a dose-related marked increase in serum DHEA and DHEA-S was observed. Serum testosterone was also increased in DHEA-treated animals, while no significant changes in serum estradiol levels were detected. As in the monkey, the histology of the female rat mammary gland remained unchanged following long-term treatment with any of the two doses of DHEA.     

Pediatric germ cell tumors and parental infertility and infertility treatment: a Children's Oncology Group report

Background

Few risk factors have been established for childhood germ cell tumors (GCT). Parental infertility and infertility treatment may be associated with GCT development but these risk factors have not been fully investigated.

Methods

A case–control study of childhood GCT was conducted through the Children's Oncology Group (COG). Cases, under the age of 15 years at diagnosis, were recruited through COG institutions from January 1993 to December 2002. Controls were obtained through random digit dialing. Information about infertility and infertility treatment along with demographic factors was collection through maternal interviews. Subgroups created by gender, age at diagnosis, and tumor location were examined separately. Statistical analysis was performed using multivariate logistic regression models.

Results

Overall, no association between GCT and infertility or its treatment was found. In subgroup analysis, females whose mothers had two or more fetal losses were found to be at increased risk for non-gonadal tumors (Odds ratio (OR) = 3.32, 95% Confidence interval (CI) = 1.12–9.88). Younger maternal age was associated with a lower risk of gonadal GCT in females (OR = 0.52, 95% CI = 0.28–0.96). There was an increased risk of all GCT and gonadal GCT in males born to older mothers (OR = 2.88, 95% CI = 1.13–7.37 and OR = 3.70, 95% CI = 1.12–12.24).

Conclusion

While no association between parental infertility or its treatment and childhood GCT was found overall, possible associations with maternal age and history of recurrent fetal loss were found in subgroups defined by gender.     

Current state of practice regarding testosterone supplementation therapy in men with prostate cancer

Hypogonadal men are characterized by low serum testosterone and symptoms of low energy, decreased libido, and muscle mass as well as impaired concentration and sexual functioning. Men with prostate cancer (PCa) currently on active surveillance or post-therapy, have traditionally been excluded from management paradigms given the decade-old concern that testosterone caused PCa growth. However, there appears to be little or no relationship between serum testosterone concentration and PCa. Androgen action in the prostate has long been known to be affected by the kinetics of receptor saturation and, as such, testosterone beyond a certain baseline is unable to stimulate prostatic growth due to complete intra-prostatic androgen receptor binding. Given this physiologic concept, many clinical investigators have begun to promote testosterone supplementation therapy (TST) as safe in men with PCa. This review examines the basics of testosterone physiology and summarizes the most recent findings on the use of TST in men with PCa on active surveillance and following treatment with external beam radiotherapy, brachytherapy and radical prostatectomy.     

Social dominance and serum testosterone concentration in dyads of male Macaca fascicularis

The relationship between social dominance and serum testosterone concentration was evaluated in 24 male Macaca fascicularis in 14 dyads at 2-week intervals over an 8-month period. No associations between testosterone concentration and dominance ranks were found in dyads with "clear dominance" relationships. A significant positive association was found between testosterone concentration and dominance ranks in dyads that exhibited "contested dominance" or dominance reversals. Thus, higher testosterone concentration corresponded to social dominance in subjects dominant as the result of a contest, rather than a consequence of high relative rank.     

Decreased serum testosterone concentration in male heroin and methadone addicts

The serum concentrations of testosterone, estradiol-17β, FSH and LH were measured in 22 male subjects addicted to heroin or methadone. Serum testosterone concentration was decreased in many of these subjects without consistent abnormalities in the other hormones. It is suggested that decreased sexual function in male addicts may be partially due to a decrease in serum testosterone concentration.     

Morphological changes and EGF expression in the granular convoluted tubule cells of submandibular glands of Trypanosoma cruzi infected rats

We have previously demonstrated in rats that Chagas' disease affects the salivary glands, by promoting an enlargement of the submandibular gland. In order to further investigate possible functional alterations on infected submandibular glands, the objective of the present study was to analyze epidermal growth factor (EGF) expression on rat submandibular glands during Trypanosoma cruzi infection. Results demonstrated that infected rats presented lower levels of testosterone, and morphological changes in the granular convoluted tubule (GCT) cells of the submandibular glands, along with acinar enlargement and delayed ductal maturation at the developing granular ducts. Immunohistochemistry analysis additionally showed that only few cells immunolabelled with anti-EGF on infected rats during the acute phase of Chagas' disease, while after 64 and 90 days (chronic phase) of infection, EGF expression was similar to non-infected rats. The present findings suggest that at the acute phase of Chagas' disease, lower levels of testosterone may lead to a delayed maturation of GCT, which positively correlates with decreased EGF production by submandibular glands cells.     

Serum FSH, LH and testosterone in the male rhesus following prostaglandin injection.

Adult male rhesus were treated with PGE2, PGF2 alpha or the 13,14-dihydro-15-keto metabolite of PGE2 in a randomized crossover design. Serum concentrations of FSH, LH and testosterone were determined and compared to the respective values in the same uninjected animals. No significant changes were noted in controls or following the metabolite injection. FSH increased gradually for 4 hours after metabolite treatment. In contrast, injection of PGF2 alpha was followed by an abrupt (within 15 minutes) increase in LH and testosterone. FSH increased gradually in 2 of 3 treated animals. Injection of PGE2 was followed by a similar abrupt increase in LH concentration. This was not always associated with a significant increase in testosterone or FSH. These results demonstrate that injections of PGE2 or PGF2 alpha can change serum gonadotropin and testosterone concentrations in male rhesus monkeys, and that the effects of these two prostaglandins are qualitatively different.     

Epidermal growth factor-like material in rat submandibular gland.

By using an antiserum specific for mouse epidermal growth factor (EGF), only the granular convoluted tubule (GCT) cells revealed immunochemical staining in rat submandibular glands. There was no regular sexual difference in the frequency or size of immunoreactive cells. Extracts of gland contained an antigen which showed a complete cross-reactivity with mouse EGF in radioimmunoassays. The relative amounts of EGF, determined by a heterologous radioimmunoassay, were not significantly different in the glands of rats of the two sexes. Administration of testosterone caused an increase, in both sexes, in the number of GCT cells stained for EGF and in the amount of EGF in the gland. There was no significant sexual differeence in these two parameters after androgen treatment.     

Luteinizing hormone release by gonadotropin releasing hormone before and after castration in bulls

The magnitude of gonadotropin releasing hormone (GnRH) induced lutei nizing hormone (LH) release prior to castration, following castration, a nd during testosterone replacement in males, was compared, using 6 9-mon th-old Holstein bulls. Also, the effects of castration and testosterone replacement on patterns of episodic changes in serum LH were studied. Blood samples were collected at hourly intervals for 24 hours prior to castration, at 21 days after castration, and at 23 days postcastration a fter testosterone, 20 mg thrice daily, has been given for 24 hours. Each animal was given GnRH, 40 mcg iv, at 24 hours before castration, at 7 and 14 days after castration, and at 28 days postcastration following 6 days of testosterone treatment. GnRH caused LH release before and after castration. The LH increase was 2.5-fold at 14 days postcastratio n. Testosterone replacement did not reduce the magnitude of LH response to GnRH to precastration levels. The number of episodic increases in serum LH prior to castration averaged 3.7 daily and increased to 6.5 daily at 21 days after castration (p less than .05). The magnitude of increase in LH concentration in these epidsodic events was not affected by castration. Testosterone replacement failed to restore either the average number or change the magniture of LH increase above precastratio n levels. It was shown that LH is normally released episodically in bulls. The peaks of LH release were followed by increased testosterone in serum. Results suggest that LH release in bulls is controlled by gonadic factors other than testosterone.     

Serum testosterone, male dominance, and aggression in captive groups of vervet monkeys (Cercopithecus aethiops sabaeus)

The relationship of serum testosterone concentration to male dominance rank and frequency of aggression was investigated in stable vervet monkey social groups, each containing two or three adult males, several adult females, and their offspring. Dominance relationships were determined by noting an animal's success in intermale aggressive encounters. A striking finding was the marked within-subject variation in testosterone concentration: 5- to 10-fold fluctuations were often observed on successive days. When all 15 groups were considered together, testosterone concentration was unrelated to dominance rank. Although mean testosterone concentration for all dominant males was higher than the mean for all subordinate males, this difference was not significant. In a subset of 4 groups, the rate of aggression initiated was significantly correlated with same-day testosterone in dominant but not in subordinate males.     

Pituitary desensitization and the regulation of pituitary gonadotropin-releasing hormone (GnRH) receptors following chronic administration of a superactive GnRH analog and testosterone

We recently demonstrated that chronic daily administration of a superactive GnRH analog to intact rats resulted in an initial stimulation of serum LH levels with a subsequent return of LH levels to baseline at a time when testosterone levels were marked decreased. These data demonstrated pituatary desensitization following chronic GnRH analog treatment. Administration of GnRH analog with a dose of testosterone which did not markedly lower serum LH levels when administered alone prevented the stimulation of LH secretion by analog. The present studies were undertaken to determine the effects of GnRH analog and testosterone administration on the regulation of pituitary GnRH receptors. Pituitary GnRH receptor binding was increased by analog treatment alone at 20 days and returned to control levels at 40 and 60 days of treatment in parallel to the observed changes in serum LH, demonstrating that one mechanism by which chronic GnRH analog treatment leads to pituitary desensitization is down-regulation of pituitary GnRH receptors. Testosterone administration alone decreased pituitary GnRH receptor binding. Combined GnRH analog and testosterone administration prevented the increase in pituitary GnRH receptors observed with analog administration alone. These studies demonstrate that changes in pituitary GnRH receptor binding correlate with changes in serum LH and that the stimulatory effects of analog administration on LH are sensitive to inhibition by small doses of testosterone.     

Hyperandrogenism Due to a Testosterone-Secreting Sertoli-Leydig Cell Tumor Associated With a Dehydroepiandrosterone Sulfate-Secreting Adrenal Adenoma in a Postmenopausal Woman: Case Presentation and Review of Literature

ObjectiveTo report a case of hyperandrogenism attributable to the presence of an adrenal adenoma secreting dehydroepiandrosterone sulfate (DHEA-S) and an ovarian Sertoli-Leydig cell tumor secreting testosterone in a postmenopausal woman.MethodsThe laboratory, radiologic, and pathologic findings in our case are described. In addition, the pertinent literature is reviewed.ResultsA 56-year-old woman presented with a history of gradual increase in facial and body hair, scalp hair loss, male pattern baldness, and deepening of her voice, beginning a few years after spontaneous menopause at age 49 years. She had hypertension, obesity, and type 2 diabetes mellitus. Laboratory tests showed elevated levels of total testosterone (348 ng/dL) and DHEA-S (2,058 μg/dL), and a left adrenal tumor (3 by 4 cm) was detected on abdominal computed tomographic scan. Laparoscopic left adrenalectomy was performed, and the pathologic diagnosis was adrenal adenoma. The DHEA-S returned to normal levels, but the serum testosterone concentration remained elevated. Transvaginal ultrasonography disclosed an ovarian tumor. Bilateral oophorectomy was performed, and an ovarian Sertoli-Leydig cell tumor was diagnosed. The hormonal and clinical picture normalized after this surgical intervention.ConclusionAfter extensive review of the literature, we believe that this is the first reported case of a coincidental DHEA-S-secreting adrenal adenoma and a testosterone-secreting ovarian Leydig cell tumor causing signs of virilization. (Endocr Pract. 2009;15:149-152)