Free Testosterone Concentrations Research Articles

Interactions Between Reproductive Hormones and Neuro-Vascular Regulation During the Cold Pressor Test in Young Females

Recently, we reported that muscle sympathetic nerve activity (MSNA) is not predictive of vascular nor blood pressure outcomes during the cold pressor test (CPT) in young healthy females. Instead, we observed substantial variability in vascular responses to the CPT which ranged widely from decreases in blood flow and vascular conductance to increases in these same outcomes. Interindividual differences in circulating reproductive hormones may account for this female-specific variability in vascular responses to acute elevations in MSNA. Therefore, the purpose of this study was to test the hypothesis that reproductive hormones account for variability in MSNA, femoral blood flow (FBF), and femoral vascular conductance (FVC) during the CPT in females. We retrospectively assessed data from 9 young healthy females (age: 24±3 years) in the low hormone phase of their menstrual cycle (days 1-5). Intravenous blood samples were obtained and prospectively analyzed to assess concentrations of circulating reproductive hormones (radioimmunoassay). Integrated peroneal MSNA (microneurography; NeuroAmpEX, ADI) was quantified at rest and during a 3-minute hand CPT. Mean arterial pressure (MAP) was measured beat-by-beat (Finometer Midi, FMS) and duplex vascular ultrasound was used to measure FBF; FVC was quantified as FBF/MAP (mL/min/mmHg). All variables are expressed as the difference between the average CPT value and the average baseline value. CPT-induced changes in MSNA burst incidence (bursts/100hb) were not related to [17-beta-estradiol] (R2=0.05; P=0.60), [progesterone] (R2=0.11, P=0.47), or [free testosterone] (R2=0.01, P=0.81). Similarly, changes in FBF and FVC were not related to [17-beta-estradiol] (FBF: R2=0.04, P=0.67; FVC: R2=0.01, P=0.80) or [progesterone] (FBF: R2=0.04, P=0.67; FVC: R2=0.09, P=0.51). However, there was a negative correlation between [free testosterone] and the changes in both FBF (R2=0.69, P=0.02) and FVC (R2=0.66, P=0.03). In other words, females with higher free testosterone demonstrated larger decreases in FBF and FVC during the CPT relative to those with lower free testosterone concentrations. These data suggest that circulating free testosterone may act to increase the vasoconstrictive consequences of MSNA in females, and may account for the previously observed variability in vascular responses to MSNA during acute stress. This project was funded by: NSERC and FRQ-S. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

SHBG gene polymorphisms and their influence on serum SHBG, total and free testosterone concentrations in men.

Genetic variation in sex hormone-binding globulin (SHBG) structure may affect estimates of sex steroid exposure by altering the affinity of the protein for its ligand. Consequently, free hormone calculations assuming constant binding affinity may, for certain genetic variations, lead to incorrect diagnoses if genetic variation is not taken into consideration. To investigate the effects of genetic variation in SHBG on calculated and measured serum free testosterone (T) in men. Population-based sibling-pair study in 999 healthy men aged 25 to 45 (mean: 34.5) years. Genotyping using microarray (Illumina®) for SNPs suggested to affect binding affinity and/or concentration of SHBG or T. SHBG concentrations were measured using immunoassay and in a subset (n = 32) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Total T was measured using LC-MS/MS. Free T was calculated and in a subset (n = 314) measured directly using LC-MS/MS after equilibrium dialysis. Allelic frequencies of analyzed SNPs ranged from 0.5% to 58.2%. Compared to wild-type, SHBG concentrations were lower in rs6258 heterozygotes (-24.7%; p < 0.05) and higher in rs6259 heterozygotes, rs727428 homozygotes, and carriers of rs1799941 (+10.8 to 23.1%; all p < 0.05). Total T was higher in rs727428 homozygotes and carriers of rs5934505, rs1799941and rs6259 (+3.9 to 21.4%; all p < 0.05). No clear effects on measured free T were found, except for a trend towards higher values in rs6259 homozygotes, significant for calculated free T (+18.7%; p < 0.05) in the larger global study population. In these men, analyzed SNPs were relatively prevalent and affected serum concentrations of total T and SHBG but not calculated or measured free T except for a higher trend in rs6259 homozygotes.

The influence of body composition on the response to dynamic stimulation of the endocrine pituitary-testis axis.

Testosterone treatment is generally not recommended in men with obesity induced low serum testosterone. However, distinguishing this condition from overt testosterone deficiency in men with obesity where treatment should be initiated is a diagnostic challenge and tools to differentiate these conditions are scarce but could be of important clinical relevance. To investigate the association between body composition and dynamic responses of the pituitary-testis axis in men. Single-center cross-sectional study including 112 healthy men. Participants went through a full biochemical assessment of the pituitary-testis axis, and dynamic stimulatory tests of luteinizing hormone (LH) secretion (gonadotropin-releasing hormone (GnRH)-test) and testosterone secretion (choriogonadotropin (hCG)-test). A subset (N = 78) further had a DXA-scan performed. A higher body mass index (BMI) was associated with lower basal serum LH (BU = -0.44, 95% CI: -0.88--0.01, p = 0.04). The GnRH-stimulated LH increase was not significantly associated with BMI (BU = -0.10, 95% CI: -0.72-0.51, p = 0.74). Furthermore, a high BMI was associated with low basal testosterone (BU -0.02, 95% CI: -0.03--0.02, p < 0.001), and free testosterone (BU -15.0, 95% CI: -19.9--10.0, p < 0.001) and men with overweight and obesity had significantly lower testosterone (9%, p = 0.003 and 24%, p < 0.001) and free testosterone (25%, p = 0.006 and 50%, p < 0.001) concentrations compared to men with normal weight. The HCG-stimulated testosterone increase was significantly less dependent on BMI compared to the influence of BMI on basal testosterone concentrations (p = 0.04 for the interaction). Dynamic sex hormone responses following pituitary-testis axis stimulation were less dependent on BMI, compared to the influence of BMI on basal hormone concentrations and could potentially assist clinical decision making in patients with obesity suspected of testosterone deficiency.

The Search for the Causes of Common Hyperandrogenism, 1965 to circa 2015.

From 1965-2015, immense strides were made into understanding the mechanisms underlying the common androgen excess disorders, premature adrenarche and polycystic ovary syndrome (PCOS). The author reviews the critical discoveries of this era from his perspective investigating these disorders, commencing with his early discoveries of the unique pattern of plasma androgens in premature adrenarche and the elevation of an index of the plasma free testosterone concentration in most hirsute women. The molecular genetic basis, though not the developmental biologic basis, for adrenarche is now known and 11-oxytestosterones shown to be major bioactive adrenal androgens. The evolution of the lines of research into the pathogenesis of PCOS is historically traced: research milestones are cited in the areas of neuroendocrinology; insulin resistance, hyperinsulinism, type 2 diabetes mellitus; folliculogenesis; androgen secretion; obesity; phenotyping, prenatal androgenization, epigenetics, and complex genetics. Large scale genome-wide association studies led to the 2014 discovery of an unsuspected steroidogenic regulator DENND1A (differentially expressed in normal and neoplastic development). The splice variant DENND1A.V2 is constitutively overexpressed in PCOS theca cells in long-term culture and accounts for their PCOS-like phenotype. The genetics are complex, however: DENND1A intronic variant copy number is related to phenotype severity, and recent data indicates that rare variants in a DENND1A regulatory network and other genes are related to PCOS. Obesity exacerbates PCOS manifestations via insulin resistance and pro-inflammatory cytokine excess; excess adipose tissue also forms testosterone. Polycystic ovaries in one-quarter of apparently normal women lie on the PCOS functional spectrum. Much remains to be learned.

Clinical significance of endocrine disorders in the development of early vascular aging in males with abdominal obesity and concomitant arterial hypertension: An observational cohort study

Background. Visceral obesity is a risk factor in the development of metabolic and endocrine disorders leading to arterial hypertension and cardiovascular complications. Their early predictors include increased vascular stiffness and early vascular aging. The current literature lacks studies into the effects of changes in testosterone, cortisol, and aldosterone levels in serum on vascular stiffness and the development of early vascular aging in patients with visceral obesity. Objective. To determine the relationship between hypercortisolemia, hyperaldosteronemia, and hypotestosteronemia and vascular stiffness and the presence of early vascular aging in male patients with visceral obesity. Methods. An observational cohort study of 78 males aged 35–45 years (mean age 38.1 ± 6.5 years) diagnosed with abdominal obesity and grade 1 arterial hypertension was conducted. The mean waist circumference ranged 105.5 ± 6.9 cm; systolic and diastolic blood pressure ranged 152.5 ± 5.0 and 92.5 ± 5.0 mm Hg, respectively. The vascular age of the studied patients (n = 78) comprised 44.1 ± 6.2 years, which was statistically higher than their passport age (р &lt; 0.001). The studied patients were divided into subgroups according to both total testosterone (&lt; 12.1 nmol/l in subgroup 1A (n = 49) and ≥ 12.1 nmol/L in subgroup 1B (n = 29)) and cortisol in the evening saliva portion (&gt; 4.5 nmol/L in subgroup 2A (n = 24) and ≤ 4.5 nmol/L in subgroup 2B (n = 24)). All the patients completed the study. To assess the hormonal status, the total testosterone sex steroid-binding globulin (SSBP) and insulin in morning serum samples were investigated. Insulin resistance was assessed based on the NOMA-IR index. The concentration of total testosterone was determined by enhanced chemiluminescence (Ortho-Clinical Diagnostics, J&amp;J); the SSBP and insulin levels were determined by delayed fluorescence. Aldosterone content was determined by radioimmunoassay; free cortisol and testosterone were measured by luminescent LIA. Free and bioavailable testosterone concentrations in serum were calculated using an online calculator (issam.ch/freetesto.htm). The cardio-ankle vascular index (CAVI) was determined using a VaSera VS-15000N device, which automatically calculated the vascular age. Statistical analysis was performed using the Statistica 10.0 Windows package (StatSoft, Inc., USA). Results. The vascular age of patients with hypogonadism was statistically significantly ( р &lt; 0.001) higher than their passport age and the vascular age in males without hypogonadism. The CAVI and vascular age were also statistically significantly higher in males with functional hypercorticism (р &lt; 0.001) compared with a subgroup of patients without hypercorticism. The vascular age and CAVI increased with an increase in the salivary cortisol concentration 2200 (r = 0.5; р &lt; 0.05) and decreased with an increase in the salivary cortisol level 900 (r = –0.5; р &lt; 0.05). These parameters decreased with an increase in serum aldosterone obtained in the morning (r = –0.4; p &lt; 0.05) and increased with an increase in serum aldosterone in the evening (r = 0.4; p &lt; 0.05). In 23% (n = 18), an inversion of the daily rhythm of cortisol production was observed; in these patients, salivary cortisol levels of 2200 exceeded salivary cortisol levels of 900. The vascular age of patients with the inversion of cortisol production (49.4 ± 4.4 years) was statistically significantly ( р &lt; 0.001) different from that of patients with normal changes in salivary cortisol concentrations (41.9 ± 4.9 years). CAVI was also higher (р &lt; 0.001) in males with inverted fluctuations in salivary cortisol levels (7.51 ± 0.62) compared to those with normal diurnal rhythm (6.45 ± 0.69). The results of aldosterone evaluation revealed that 17% of the patients (n = 13) had higher aldosterone levels in the evening serum portion compared to the morning serum portion. In these patients, the vascular age (45.8 ± 5.1 years) was higher ( р &lt; 0.001) than that in males with normal physiologic changes in aldosterone levels (41.6 ± 5.7 years). A similar pattern was observed when comparing vascular stiffness indices. Thus, the CAVI in men with inverted changes in aldosterone concentration (6.9 ± 0.8) was significantly higher ( р &lt; 0.001), compared to that in men with physiological changes in blood aldosterone levels (6.4 ± 0.8). Conclusion. Endocrinologic disorders in male patients with visceral obesity and concomitant arterial hypertension (functional hypogonadism, functional hypercorticism, inverted daily rhythms of cortisol and aldosterone production) contribute to the development of early vascular aging by increasing vascular stiffness.

Sex-steroid hormones and risk of postmenopausal estrogen receptor-positive breast cancer: a case-cohort analysis.

Sex-steroid hormones are associated with postmenopausal breast cancer but potential confounding from other biological pathways is rarely considered. We estimated risk ratios for sex-steroid hormone biomarkers in relation to postmenopausal estrogen receptor (ER)-positive breast cancer, while accounting for biomarkers from insulin/insulin-like growth factor-signaling and inflammatory pathways. This analysis included 1208 women from a case-cohort study of postmenopausal breast cancer within the Melbourne Collaborative Cohort Study. Weighted Poisson regression with a robust variance estimator was used to estimate risk ratios (RRs) and 95% confidence intervals (CIs) of postmenopausal ER-positive breast cancer, per doubling plasma concentration of progesterone, estrogens, androgens, and sex-hormone binding globulin (SHBG). Analyses included sociodemographic and lifestyle confounders, and other biomarkers identified as potential confounders. Increased risks of postmenopausal ER-positive breast cancer were observed per doubling plasma concentration of progesterone (RR: 1.22, 95% CI 1.03 to 1.44), androstenedione (RR 1.20, 95% CI 0.99 to 1.45), dehydroepiandrosterone (RR: 1.15, 95% CI 1.00 to 1.34), total testosterone (RR: 1.11, 95% CI 0.96 to 1.29), free testosterone (RR: 1.12, 95% CI 0.98 to 1.28), estrone (RR 1.21, 95% CI 0.99 to 1.48), total estradiol (RR 1.19, 95% CI 1.02 to 1.39) and free estradiol (RR 1.22, 95% CI 1.05 to 1.41). A possible decreased risk was observed for SHBG (RR 0.83, 95% CI 0.66 to 1.05). Progesterone, estrogens and androgens likely increase postmenopausal ER-positive breast cancer risk, whereas SHBG may decrease risk. These findings strengthen the causal evidence surrounding the sex-hormone-driven nature of postmenopausal breast cancer.

Influence of oral hormonal contraception on the concentration of anti-Müllerian and reproductive hormones in patients with polycystic ovary syndrome

Aim: To investigate the effects of three-month use of oral hormonal contraception (OHC) on hormonal status and ovarian reserve indicator (anti-Müllerian hormone, AMH) in patients with polycystic ovary syndrome (PCOS).Methods: 19 patients with diagnosed PCOS and clinical and laboratory signs of hyperandrogenism without additional comorbidities and co-medication were included in the study. All participants received therapy with the same oral hormonal contraceptive (fixed combination of 0.035 mg ethinyl estradiol and 2 mg cyproterone acetate). The main outcomes were the concentrations of reproductive hormones measured before starting therapy and in the first cycle following therapy. Hormone concentrations were analysed using the immunochemical electrochemiluminescence (ECLIA) method.Results: Initial concentrations of total and free testosterone and AMH were elevated, while initial concentrations of other reproductive hormones were within reference values. By applying the therapy, the concentrations of AMH, luteinizing hormone (LH) and estradiol decreased by more than 20% and those of free testosterone by 85%. The concentration of sex hormone binding globulin (SHBG) increased by 44%.Conclusions: Three months of oral hormonal contraception with 35 μg ethinyl estradiol and 2 mg cyproterone acetate reduced elevated concentrations of AMH and free testosterone in PCOS patients. The decrease in serum AMH concentration indicates a temporary interruption of folliculogenesis as well as the selection of follicles from preantral to antral, and the decrease in androgens has a positive effect on the clinical condition and symptoms of patients with PCOS.

Dietary fat quality and serum androgen concentrations in middle-aged men

Background/ObjectivesAverage testosterone concentrations in men have declined over the last few decades. The reasons for this are not fully known, but changes in dietary fat quality have been suggested to have a role. This study aimed to investigate the associations of different dietary fatty acids with serum androgen concentrations.Subjects/MethodsA total of 2546 men with a mean age of 53 from the Kuopio Ischaemic Heart Disease Risk Factor Study were included in this cross-sectional study. Associations between dietary saturated (SFA), monounsaturated (MUFA), polyunsaturated (PUFA) and trans (TFA) fatty acids and concentrations of serum total and free testosterone and steroid hormone binding globulin (SHBG) were analyzed with analysis of covariance and linear regression analysis. Associations of isocaloric replacement of nutrients and androgen concentrations were analyzed with multivariate nutrient-density models.ResultsAfter adjustment for age, examination year and energy intake, higher SFA intake was associated with higher serum total and free testosterone and SHBG concentrations, and higher PUFA intake with lower concentrations. However, the associations were attenuated and not statistically significant after further adjustments for potential confounders. MUFA and TFA intakes were not associated with androgen concentrations. In isocaloric substitution models, replacing dietary protein with SFA was associated with higher serum total testosterone and SHBG concentrations. After excluding men with history of CVD or diabetes (n = 1021), no statistically significant associations were found.ConclusionsDietary fat quality was not independently associated with serum androgen concentrations in middle-aged men. However, replacing protein with SFA may be associated with higher serum androgen concentrations.

Exploring the efficacy and safety of a novel standardized ashwagandha (Withania somnifera) root extract (Witholytin®) in adults experiencing high stress and fatigue in a randomized, double-blind, placebo-controlled trial.

Stress is a state of homeostasis in the body being challenged, resulting in a systemic response. It has become more prevalent in recent years and affects mental and physical health. Evaluate the effects of ashwagandha on stress, fatigue, and sex hormones in overweight or mildly obese men and women with self-reported stress and fatigue. Two-arm, parallel-group, 12-week, randomized, double-blind, placebo-controlled trial on overweight or mildly obese men and women aged 40-75 years, supplementing with 200 mg of an ashwagandha root extract (Witholytin®) twice daily. Supplementation with ashwagandha was associated with a significant reduction in stress levels based on the Perceived Stress Scale (primary outcome); however, the improvements were not significantly different to the placebo group (p = 0.867). Based on the Chalder Fatigue Scale, there was a statistically significant reduction in fatigue symptoms in the ashwagandha group compared to the placebo group (p = 0.016), and participants taking ashwagandha also experienced a significant increase in heart rate variability (p = 0.003). However, there were no significant between-group differences in other self-report outcome measures. In the men taking ashwagandha, there was a significant increase in the blood concentrations of free testosterone (p = 0.048) and luteinizing hormone (p = 0.002) compared to the placebo group. The results of this study suggest that in overweight middle-to-older age adults experiencing high stress and fatigue, compared to the placebo, ashwagandha did not have a significantly greater impact on perceived stress levels. However, based on secondary outcome measures, it may have anti-fatigue effects. This may be via its impact on the autonomic nervous system. However, further research is required to expand on these current findings.

Stress hormone response to instrumented elective lumbar spine fusion surgery.

To understand the systemic effect of major spine surgery, we investigated stress, anabolic and catabolic hormonal levels and their association with interleukin 6 (IL-6) in patients undergoing elective lumbar spine fusion surgery. Blood samples were collected preoperatively, and at 1, 3, 42, 90days postoperatively (POD) from 49 patients who underwent elective lumbar spine fusion surgery. Serum concentration of cortisol was below the preoperative value at POD 1 but did not differ from the baseline values thereafter. Adrenocorticotropic hormone (ACTH) decreased at PODs 1 and 3. Testosterone decreased at PODs 1 and 3 in men, and at POD 3 in women. Sex hormone-binding globulin decreased at PODs 1 and 3 in both genders. No changes were observed in free testosterone or growth hormone concentrations. Insulin-like growth factor 1 increased significantly above the preoperative level at PODs 42 and 90 in women, and at POD 42 in men. IL-6 was significantly elevated at PODs 1 and 3. Increases in IL-6 from the preoperative level to POD 1 correlated significantly with decreases of cortisol at POD 1 but not with ACTH. There were only short-term stress hormonal changes after elective lumbar spine fusion surgery. Cortisol changes after elective lumbar spine surgery are transient and might be partly cytokine induced and non-ACTH driven since there was association between cortisol and IL-6 but not with ACTH and IL-6.

Combined treatment of idiopathic male infertility using low-intensity laser therapy and complex prostate peptides: single-centre experience

Introduction. The study of effective and safe combined methods of conservative treatment of idiopathic male infertility is of great interest.Objective. To evaluate the effectiveness of the combined use of low-level laser therapy (LLLT) in the infrared (IR) spectrum and bioregulatory therapy with complex prostate peptides on the semen parameters and sperm DNA fragmentation (SDNAF) in men with idiopathic infertility.Materials &amp; methods. The study is based on the results of examination and treatment of 97 men with idiopathic infertility. Prior to the start of treatment, all patients were divided into three groups by randomization. Patients in group 1 (n = 34) underwent a course of LLLT in the IR spectrum consisting of 10 procedures q.o.d. Patients in group 2 (n = 31) were prescribed complex prostate peptides Supp. Prostatilen® AC (30 + 180 mg) PR q.h.s. FXD, with a repeat of the course of treatment after 5 days. Patients in group 3 (n = 32) underwent combined treatment: LLLT sessions in the IR spectrum (10 procedures q.o.d.) and a course of Supp. Prostatilen® AC (30 + 180 mg) PR q.h.s. FXD, with a repeat of the course of treatment with peptides according to the same scheme after 5 days. Control examination of patients in all groups was carried out two months after the end of treatment.Results. As a result of treatment of patients in groups 1, 2, and 3, a significant improvement in the main semen parameters, a decrease in the grade of pathological SDNAF, an increase in serum total and free testosterone concentration (p &lt; 0.05) were observed. A comparative statistical analysis of the results obtained in three groups of patients showed that the combined treatment of patients of group 3, who were prescribed a course of LLLT in combination with complex prostate peptides Supp. Prostatilen® AC, was significantly more effective in improving concentration (p &lt; 0.01) and progressive sperm motility (p &lt; 0.01), as well as a decrease in the grade of pathological SDNAF (p &lt; 0.05), than in groups 1 and 2.Conclusion. Combined therapy that includes complex prostate peptides and LLLT in the IR spectrum improves the main semen parameters and reduces the grade of pathological SDNAF in men with idiopathic infertility.

Immunomodulatory effects of testosterone and letrozole during Plasmodium berghei ANKA infection.

Malaria is one of the leading health problems globally. Plasmodium infection causes pronounced sexual dimorphism, and the lethality and severity are more remarkable in males than in females. To study the role of testosterone in the susceptibility and mortality of males in malaria, it is common to increase its concentration. However, this strategy does not consider the enzyme CYP19A1 aromatase, which can transform it into oestrogens. To avoid the interference of oestrogens, we inhibited in vivo CYP19A1 aromatase with letrozole and increased the testosterone level by exogen administration before infection with Plasmodium berghei ANKA. We measured the impact on free testosterone, 17β-oestradiol and dehydroepiandrosterone levels in plasma; additionally, we evaluated parasitaemia, body temperature, body mass, glucose levels and haemoglobin concentration. Furthermore, we evaluated the effects of testosterone on the immune response; we quantified the CD3+/CD4+, CD3+/CD8+, CD19+, Mac-3+ and NK cells in the spleen and the plasma concentrations of the cytokines IL-2, IL-4, IL-6, IFN-, IL-10, TNF-α and IL-17A. Finally, we quantified the levels of antibodies. We found that mice treated with the combination of letrozole and testosterone and infected with Plasmodium berghei ANKA had increased concentrations of free testosterone and DHEA but decreased levels of 17β-oestradiol. As a result, parasitaemia increased, leading to severe anaemia. Interestingly, testosterone increased temperature and decreased glucose concentration as a possible testosterone-mediated regulatory mechanism. The severity of symptomatology was related to critical immunomodulatory effects generated by free testosterone; it selectively increased CD3+CD8+ T and CD19+ cells but decreased Mac-3+. Remarkably, it reduced IL-17A concentration and increased IL-4 and TNF-α. Finally, it increased IgG1 levels and the IgG1/IgG2a ratio. In conclusion, free testosterone plays an essential role in pathogenesis in male mice by increasing CD8+ and decreasing Mac3+ cells and mainly reducing IL-17A levels, which is critical in the development of anaemia. Our results are important for understanding the mechanisms that regulate the exacerbated inflammatory response in infectious diseases and would be useful for the future development of alternative therapies to reduce the mortality generated by inflammatory processes.

Endocrine disorders after combined chemoradiotherapy in Hodgkin Lymphoma survivors

Hodgkin's lymphoma (HL) is one of the most common malignant lymphoproliferative diseases. Chemotherapy and radiotherapy used in the treatment of LH induce a number of toxic effects leading to dysfunction of endocrine system. Hormonal disorders in HL and their relationships with the therapy used remain to be clarified. To assess disorders of the endocrine function of thyroid, parathyroid glands and gonads in HL survivors. Screening of endocrine dysfunction of the thyroid, parathyroid glands and gonads was performed in 160 adult patients with HL, 55 men and 105 women, at remission stage induced by chemotherapy or chemoradiotherapy. Forty healthy subjects, matched by age, were acted as control. The levels of TSH, T3, free T4, PTH, FSH, LH, free testosterone, dehydroepiandrosterone sulfate (DHEA-S), and sex-hormone binding globulin (SHBG) were measured in blood serum by ELISA. Bone mineral density (BMD) was assessed by DEXA. Hypothyroidism (25%), hyperparathyroidism (15.6%) and hypogonadism (29% of men and 25.3% of women) were the most prevalent endocrine disorders in LH survivors. Hypothyroidism was significantly more common in patients after chemoradiotherapy than in those who received only chemotherapy (χ2=9.4, р=0.002). In patients with hyperparathyroidism, there were negative correlations between PTH levels and BMD in the lumbar spine (r=-0.74, p=0.00002) and in the femoral neck (r=-0.66, p=0.0003). Men with HL demonstrated lower free testosterone concentrations when compared to control (p=0.04); LH and FSH levels were elevated (p=0.0004 and p=0.04, respectively). In men with HL the levels of DHEA-S were reduced (p=0.0009). The increased SHBG concentrations were revealed in 13 (23.6%) men. Women of reproductive age with HL had higher levels of LH in the luteal phase (p=0.05) and FSH in the follicular phase (p=0.02) than controls. The data indicate a high prevalence of the dysfunctions of thyroid, parathyroid glands, and gonads in HL survivors. Screening for endocrine disorders in these patients is highly recommended.

Training-induced impairment of endothelial function in track and field female athletes

Professional athletes are often exposed to high training loads that may lead to overfatigue, overreaching and overtraining that might have a detrimental effects on vascular health. We determined the effects of high training stress on endothelial function assessed by the flow-mediated dilation (FMD) and markers of glycocalyx shedding. Vascular examination as well as broad biochemical, hormonal and cardiometabolic evaluation of sprint and middle-distance female runners were performed after 2 months of preparatory training period and compared to age-matched control group of women. Female athletes presented with significantly reduced FMD (p < 0.01) and higher basal serum concentrations of hyaluronan (HA) and syndecan-1 (SDC-1) (p < 0.05 and p < 0.001, respectively), that was accompanied by significantly lower basal serum testosterone (T) and free testosterone (fT) concentrations (p < 0.05) and higher cortisol (C) concentration (p < 0.05). It resulted in significantly lower T/C and fT/C ratios in athletes when compared to controls (p < 0.01). Moreover, fT/C ratio were significantly positively correlated to FMD and negatively to HA concentrations in all studied women. Accordingly, the training load was significantly negatively correlated with T/C, fT/C and FMD and positively with the concentrations of HA and SDC-1. We concluded that young female track and field athletes subjected to physical training developed impairment of endothelial function that was associated with anabolic-catabolic hormone balance disturbances. Given that training-induced impairment of endothelial function may have a detrimental effects on vascular health, endothelial status should be regularly monitored in the time-course of training process to minimalize vascular health-risk in athletes.

Impact of Alpinia galanga and zinc on semen quality and some reproductive hormone constituents in California rabbit bucks.

The objective of the current study was to investigate the influence of synergism of the dry powder of Alpinia galanga rhizomes (AGR) and/or zinc sulfate in the diet on semen quality and reproductive traits of California rabbit bucks. The study was conducted in two stages. First stage: appreciation of semen characteristics, 36 California rabbit bucks (aged 5 months) with average body weights of 2980 g were divided randomly into six treatments (six individuals each). The treatment groups were: first group, control fed basal diet (C); second group, fed basal diet plus 1 g AGR/kg dry matter (DM) (AGR1); third group, fed basal diet plus 2 g AGR/kg DM (AGR2); fourth group, fed basal diet plus 200 mg Zn/litre drinking water (Zn); fifth group, fed basal diet plus 1 g AGR/kg DM and 200 mg Zn/litre drinking water (AGR1 + Zn); sixth group, fed basal diet plus 2 g AGR/kg DM and 200 mg Zn/litre drinking water (AGR2 + Zn). Second stage: the previous bucks were used to determine the efficiency of semen on reproductive fertility traits, 48 mature does (aged 6 months, nulliparous) with an average body weight of 3050 ± 20.7 g were divided randomly into six treatments and inseminated with previous groups of treated bucks. The results of the first stage, recorded high activity on gonadotropins hormones: follicle-stimulating hormone (FSH) and luteinizing hormone (LH), free testosterone (FT), progesterone (P4) and oestrogen (E217β) concentrations for AGR1 + Zn and AGR2 + Zn compared with the control group. Groups AGR1, AGR2, AGR1 + Zn and AGR2 + Zn had significantly lowered concentrations of triglycerides, total cholesterol, low-density lipoprotein, and malondialdehyde (MDA), whereas high-density lipoprotein and total antioxidant capacity (TAC) were increased significantly compared with the control group. The group supplemented with AGR with or without Zn had significantly improved ejaculate volume, advanced motility, sperm concentration, and cell integrity. Fertility rate and litter size were improved in all groups compared with the control. It was concluded that supplementing diets with Alpinia galanga and Zn significantly increased sperm percentage, motility and reproductive hormones (testosterone, FSH, LH, E217β, P4). This suggested that this plant when used may be favourable for improved sperm quality and fertility parameters.

An antibody-free LC-MS/MS method for the quantification of sex hormone binding globulin in human serum and plasma.

Sex hormone binding globulin (SHBG) is a hormone binding protein which plays an important role in regulating the transport and availability of biologically active androgens and estradiol to target cells and used to calculate free testosterone concentrations. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed, featuring an albumin removal step followed by a tryptic digestion. After a reduction step with dithiothreitol and alkylation with iodoacetamide three signature peptides were used for the quantification of SHBG. The method enables the quantification of serum and plasma SHBG over the clinically relevant range of 200-20,000ng/mL and was validated according to the mostrecent guidelines. The LC-MS/MS method correlates wellwith the Abbott Alinity immunoassay (R2>0.95), but theLC-MS/MS results are on average 16-17% lower than the immunoassay results, which is consistent for all three signature peptides. The LC-MS/MS method which includes an albumin depletion step allows quantification of SHBG in serum and plasma without an immunocapture step at clinically relevant SHBG levels, thus contributing to better lab-to-lab consistency of results.

Connection of Low Serum Testosterone Levels in Cardiovascular Disease in Metabolic Syndrome Patients with Diagnosis of Critic Iliac Artery Stenosis (TASC II A and B) and Can Exercise Improve those Levels and Primary Potency of Revascularization after Surgical and Endovascular Treatment? (Pilot Study)

Introduction: Low concentration of free testosterone and low serum testosterone levels are associated in males patients with metabolic syndrome and cardiovascular disease and in some cases in connection with critic iliac stenosis and also in most cases with lack of physical activity. Exercise can be also important factor to improve primary bypass and endovascular potency and general health benefits after surigal and endovascular treatment in patients with critical ishemia over 75% of lumen obstruction of short liiac artery segment have positive impact on improvemt of generely Testosterone Levels. Subjects and Methods: 72 selected Patients with Cardiovascular Disease with Metabolic Syndrome and Critic Iliac Artery Stenosis (TASC II A and B) were observed in period 3 years between ferbruary 2015 and ferbruary 2018. In focus was 1 year potency after invasive and minimalinvasive tertmant (45 patients treated with surgical dacron reconstruction and 25 patients with endovascular tretmant of short segment critical iliac artery stenosis (TASC II A and B). Results: In the total population no difference in changes of CIMT from baseline was observed between the standard exercise group and controls. However, there was no significant interaction between the effect of exercise training and primary bypass potency during 1 year. Conclusion: Primary potency of Endovascular procedure and Dacron bypass revascularization after 1 year in exercise training group significantly rise of testosterone levels higher but still not therapy adequate. With potential optimization testosterone therapy levels we can answer of the question. Is any major influence on primary bypass potency and are higher levels of free testosterone are preventing further progressing of cardiovascular disease and generel symptomatic and asymptomatic atherosclerosis?

PS-BPC07-2: EFFECTS OF MINERALOCORTICOID RECEPTOR ANTAGONISTS ON SEX HORMONES AND BODY COMPOSITION IN PATIENTS WITH PRIMARY ALDOSTERONISM

Objective: Mineralocorticoid receptor antagonists are frequently used for the treatment of primary aldosteronism. Steroidal mineralocorticoid receptor antagonists may have antagonistic actions on androgen receptors, agonistic actions on progesterone receptors, and antagonistic actions on mineralocorticoid receptors. Because anti-androgen effects may cause body fat accumulation and skeletal muscle atrophy, there are concerns that this drug may have adverse effects on body composition. Design and method: Therefore, in this randomized prospective study, we compared the adverse effects of spironolactone, a steroidal mineralocorticoid receptor antagonist, and esaxerenone, a nonsteroidal mineralocorticoid receptor antagonist, on sex hormone levels and body composition in patients with primary aldosteronism without severe renal dysfunction. Results: The serum concentration of free testosterone was significantly higher in the spironolactone group than in the esaxerenone group in both males and females. However, the levels of estradiol, progesterone, luteinizing hormone, and follicle-stimulating hormone did not significantly increase. Changes in body fat percentage and muscle mass rate were not significantly different between the two groups. No patient showed a serum potassium level &gt; 6.0 mEq/L; however, serum potassium levels were significantly higher in the spironolactone group than in the esaxerenone group. Conclusions: These data indicate that spironolactone may have antagonistic effects on androgen receptors. Esaxerenone did not show any apparent adverse effects, suggesting that it can be safely used in patients with primary aldosteronism.

Acute Hormonal Responses to Multi-Joint Resistance Exercises with Blood Flow Restriction.

The purpose of this study was to investigate the acute effects of multi-joint resistance exercises (MJRE) with blood flow restriction on hormonal responses. Ten men participated in the study and underwent two experimental protocols in random order: four sets (30, 15, 15, and 15 reps, respectively) of MJRE (half squat and horizontal chest press) were performed with 20% of 1RM and a rest time between sets of 30 s, combined with intermittent blood flow restriction (LI + BFR protocol); and four sets (8, 8, 8, 20 reps, respectively) of the same MJRE performed with 75% of 1RM load (HI protocol), with a 90 s rest between the first three sets and 30 s between the third to the fourth set. Blood samples were collected before (PRE), immediately after (POST), and 15 min after the performance of MJRE (POST15). A time effect was observed for growth hormone (GH) and insulin-like-growth-factor-1-binding-protein-3 (IGFPB-3), but no protocol effects or interactions between protocol and times were observed (p > 0.05). There was no effect of either protocol or time (p > 0.05) on total testosterone, free testosterone, or cortisol concentrations. However, significant (p < 0.05) increases were observed in the GH serum concentrations of 2072.73% and 2278.5%, HI, and LI + BFR protocols, respectively, from the PRE to POST15 test. In addition, there was an increase of 15.30% and 13.29% in the IGFPB-3 concentrations (p < 0.05) from PRE to POST0 times for HI and LI + BFR protocols, respectively. Furthermore, there was a decrease of −6.17% and −11.54%, p = 0.00, between the times POST0 to POST15 in the IGFPB-3 for the HI and LI + BFR protocols, respectively. It is concluded that multi-joint resistance exercises combined with intermittent blood flow restriction seemed to promote acute hormonal responses in a manner similar to traditional exercise with high loads. Future studies may investigate whether chronic use of LI + BFR with MJRE may promote muscle hypertrophy.

ODP419 Measurement of Free Testosterone in Serum using Equilibrium Dialysis Coupled with ID-UHPLC-MS/MS: in Comparison to Calculated FT

Abstract Assessment of free testosterone (FT) has been recommended in recent clinical practice guidelines as a biomarker for diagnosis and management of testosterone related disorders such as male testosterone deficiency due to hypogonadism and female androgen excess due to polycystic ovary syndrome (PCOS). FT is currently measured using direct commercial immunoassays, assays using equilibrium dialysis (ED) or other separation techniques, or estimated by calculating free testosterone (cFT) using different formulas. The most widely used formula was developed by Vermeulen et al and uses total testosterone and sex hormone binding globulin (SHBG) concentrations. The ED approach is recognized as the gold standard; however, it is technically challenging for routine clinical use. CDC's Clinical Standardization Program (CDC CSP) has developed a high throughput method using ED coupled with isotope dilution ultra-high-performance liquid chromatography tandem mass spectrometry. The study to evaluate the agreement of this ED-based method to the cFT using different algorithms is being conducted. Serum samples are dialyzed in a custom-designed multi-well plate against a protein-free HEPES buffer (pH 7.4) at 37 °C until equilibrium. After isolating endogenous FT from protein-bound testosterone by ED, isotope-labeled internal standard (13C3-testosterone) was added to the resulting dialysate for quantification. Certified pure primary reference material (National Measurement Institute-M914) was used to prepare calibrators, enabling traceable quantitation and ensuring measurement trueness. FT was further isolated from the dialysate matrix using supported liquid extraction and a chromatographic separation from interfering compounds and quantitation by tandem MS. The assay allowed detection of 2.9 pg/mL FT, with the bias within ± 5% and precision less than 10% CV. A total of 45 samples with a wide range of TT (21-912 ng/dL) and SHBG (11-129 nmol/L) were analyzed. The obtained FT values were compared to the cFT values obtained with the Vermeulen algorithm. The mean difference of 34.1% (95% CI 29.1% to 39.9%) between the measured and calculated FT were observed, with majority of the calculated values overestimating FT concentrations. The described high throughput method for FT allows for sufficiently accurate and precise measurement and can provide more reliable FT measurements compared to cFT (that can lead to overestimation of FT) for routine applications including large epidemiologic studies. Presentation: No date and time listed