BACKGROUND

Stanozolol is a synthetic derivative of dihydrotestosterone (DHT), and one of the frequently detected anabolic steroids in doping analysis. The current work describes the development and validation of the stability-indicating TLC-densitometric method for sensitive and specific estimation of stanozolol even its degradation product being there. Precoated silica gel TLC aluminium plates were utilized as the stationary phase and the eluent comprised of petroleum ether: acetone (6:4, v/v). Densitometric analysis of stanozolol was achieved at λmax 750 nm in the absorbance mode after staining with phosphomolybdic acid (PMA). Stress degradation of stanozolol was carried out under various reaction conditions including acid, base and neutral hydrolysis, wet and dry heating treatment, oxidation, and photo-degradation. Resulted stress samples and pharmaceutical products were analyzed with the developed TLC method.

RESULTS

This system showed a compact spot for stanozolol at Rf value of 0.46 ± 0.02. The data of linear regression analysis indicated a good linear relationship over the range of 200-1200 ng/spot concentrations. The method was validated for robustness, precision and recovery. The LOD and LOQ were 1.6 and 5.1 ng/spot, respectively. Under various stressed conditions, stanozolol showed degradation only under acidic hydrolysis. Peak of a degraded product was well resolved from the stanazolol with reasonably different Rf value and identified as 17, 17-dimethyl-l8-nor-5α-androst-13(14)-eno [3,2c] pyrazole through 1D- and 2D-NMR spectroscopic techniques and ESI-QqTOF-MS/MS analysis.

CONCLUSIONS

Result reflected that the stanozolol is majorly affected by the acidic condition. Statistical analysis indicated the application of the developed stability-indicating TLC-densitometric method for routine analysis of stanozolol in the presence of its degradation product.

<em>5α</em>-R isozymes (types 1 and 2) play an important role in prostate gland development because they are responsible for intraprostatic dihydrotestosterone (<em>DHT</em>) levels when the physiological serum testosterone (T) concentration is low. In this study, we synthesized seven novel dehydroepiandrosterone derivatives with benzimidazol moiety at C-17, and determined their effect on the activity of <em>5α</em>-reductase types 1 and 2. The derivatives with an aliphatic ester at C-3 of the dehydroepiandrosterone scaffold induced specific inhibition of <em>5α</em>-R1 activity, whereas those with a cycloaliphatic ester (cyclopropyl, cyclobutyl, or cyclopentyl ring) or an alcohol group at C-3 inhibited the activity of both isozymes. Derivatives with a cyclohexyl or cycloheptyl ester at C-3 showed no inhibitory activity. In pharmacological experiments, derivatives with esters having an alcohol or the aliphatic group or one of the three smaller cycloaliphatic rings at C-3 decreased the diameter of male hamster flank organs, with the cyclobutyl and cyclopentyl esters exhibiting higher effect. With exception of the cyclobutyl and cyclopentyl esters, these compounds reduced the weight of the prostate and seminal vesicles.

A novel photocatalysis to construct the 3-acyl-4-arylcoumarin framework from simple aldehyde with ynoate is described. The reaction proceeded through an acyl radical intermediate generated by hydrogen atom abstraction from aldehyde, followed by reaction with ynoate and then cyclization to afford coumarins. This valuable radical cyclization reaction gave over 20 coumarin derivatives in moderate to good yields with inexpensive 2-tBu-anthraquinone as a catalyst. In addition, synthetic coumarins were investigated for <em>5α</em>-dihydrotestosterone (<em>DHT</em>)-induced secretion of prostate-specific antigen (PSA) levels and cell proliferation of androgen-dependent CWR22Rv1 cells.

OBJECTIVE

The present study was aimed to evaluate the time-mannered and dose-dependent effects of <em>5α</em>-dihydrotestosterone (<em>5α</em>-<em>DHT</em>) on the proliferation and differentiation of bone forming cells using MC3T3-E1 cells.

METHODS

Cell proliferation was analyzed using MTS and phase contrast microscopic assays. Osteogenic differentiation was assessed through a series of in vitro experiments including crystal violet staining, alkaline phosphatase (ALP) activity, and Van Gieson (VG) staining. Taken together, the efficiency of bone mineralization was examined by using alizarin red s (ARS) staining, Von Kossa staining, scanning electron microscopy (SEM) and energy dispersive x-ray (EDX) analysis.

RESULTS

The resulting data revealed that <em>5α</em>-<em>DHT</em> exhibits promising potential particularly at a dose of 0.1 ng/ml, in promoting the growth of MC3T3-E1 cells compared to the control group (CN). Moreover, a significantly higher ALP activity was evident in the experimental group treated with <em>5α</em>-<em>DHT</em> compared to the CN group at various time intervals. MC3T3-E1 cells treated with <em>5α</em>-<em>DHT</em> also expressed a remarkably higher collagen deposition and mineralization (calcium and phosphate contents) compared to the CN group at various time intervals.

CONCLUSIONS

Conclusively, we suggest that <em>5α</em>-<em>DHT</em> exhibits outstanding potential of promoting proliferation and differentiation in osteoblasts which could be the in vitro basis for the efficacy of <em>5α</em>-<em>DHT</em> in the treatment of androgen-deficient male osteoporosis.

(Note: This unorthodox paper contains the first argument for heart disease being a programmed age change and promoted by the dramatic, post age-40 increases in the hormones FSH and hCG seen in some individuals.) A recent issue of Science suggests that the evolutionary purpose of sex is unknown.

UNASSIGNED

Surviving to adulthood implies a valuable gene combination which is destroyed by sexual recombination. This should be detrimental to offspring. PROPOSED: Sex is group-selected in prey to allow coalescence of beneficial, and disposal of detrimental, mutations in single individuals enabling rapid adaptation to novel predation. Group selection is a universal force driven by local inter-species (not intra-species) competition. Aging, metabolism, litter size, and fixed body size are directly linked. Sexual recombination and chromosomes destroy gene linkage and exist because mutations are usually detrimental, rarely positive, and occur in linked groups. In unevolving environments, sex is selected against and asexuality emerges. Periodic evolution of novel predators, like man, can explain the 'punctuated equilibria' fossil record. Genes inhibited by methylation or chromatin condensation, expressed at older ages in predation-minimized environments, allow for group selection. Stress increases mutation rates and beneficial mutation likelihood. Females select bigger, brighter, louder, or stronger males that can survive predator attention. Size approximates age and thus predator encounters; male traits represent predation-survival potential. Human male traits include, balding, acne, beard-length, wrinkling, graying, nose/ear growth. Progeria accelerates development of most male traits. Domination of groups by single males allows rapid predation-defense evolution: adolescent males are expelled, brave the wild, and expel another group's male to mate. If expelled and dominant males are culled by predation, males reaching puberty first will reproduce. Hormonal acceleration of puberty accelerates aging/population turnover, induces smaller bodies, larger litters. With a fixed group biomass, more, smaller, stressed individuals with faster aging/turnover, increase beneficial mutation likelihood. 'Kin selection', where dominant families are supported by celibate relatives, allow the best group genes to survive famine. Dominant families gorge while others starve. Equal food sharing results in group extinction leading to group-evolved human traits of social hierarchy, greed, king/queen/God worship. Menstrual hormone cycling parallels aging. FSH and <em>DHT</em> promote ovarian, hair, acne, dental, and arterial follicle development causing ovulation, hair growth, pimples, dental caries, and atherosclerotic soft plaques. Soft plaques contain macrophages and LDL plug; upper plaque layers thin and rupture, releasing LDL plug, causing thrombosis. FSH withdrawal or LH/hCG increases trigger ovulation and thrombosis. Artery narrowing atherosclerotic hard plaques are stress-induced through cortisol-promoted necrotic calcification. LH/hCG-induced apoptosis promotes ovulation and aging-related somatic atrophy. Long-term estradiol stimulates, while progesterone suppresses, gonadotropin levels. Estradiol protects by inhibiting gonadotropin bioactivity and has extracellular antioxidant, but intranuclear free radical, effects. Female X-linked gene mosaicism conserves evolved aging systems. Maternal age factors for chromosomal trisomy suggest menopause prevents human parthenogenesis. Homosexuality and serial killing inhibit genetic contribution by individuals evolutionarily perceived as stressed. Smoking during pregnancy may induce homosexual offspring. Nitric oxide, a free radical, stimulates cGMP, but not cAMP. cGMP likely first evolved as an antioxidant defense to free radicals. Human aging syndromes might reflect human evolution progression. AS#4 affects tissues evolved from plant ancestors, AS#<em>5a</em> - from predators, AS#5b-immune system, and AS#6-sex tissues. (ABSTRACT TRUNCA

BACKGROUND

Alopecia X in dogs is a noninflammatory alopecia that may be caused by a hormonal dysfunction. It may be similar to androgenic alopecia in men that is caused by the effect of dihydrotestosterone (<em>DHT</em>). The <em>5α</em>-reductase isoenzymes, <em>5α</em>R1 and <em>5α</em>R2, and a recently described <em>5α</em>R3, are responsible for the conversion of testosterone into <em>DHT</em>. However, which <em>5α</em>-reductases are present in canine skin has not yet been described.

OBJECTIVE

The main objective of this study was to determine the pattern of expression of <em>5α</em>-reductase genes in canine skin.

METHODS

Skin biopsies were obtained from healthy, intact young-mature beagles (three males, four females) at three anatomical sites normally affected by alopecia X (dorsal neck, back of thighs and base of tail) and two sites generally unaffected (dorsal head and ventral thorax). Prostate samples (n = 3) were collected as positive controls for <em>5α</em>-reductase mRNA abundance measurement by real-time PCR.

RESULTS

We detected mRNA encoding <em>5α</em>R1 and <em>5α</em>R3 but not <em>5α</em>R2. There were no significant differences in <em>5α</em>R1 and <em>5α</em>R3 mRNA levels between the different anatomical sites, irrespective of gender (P>> 0.05). Moreover, the mean mRNA abundance in each anatomical site did not differ between males and females (P>> 0.05).

CONCLUSIONS

To the best of the authors' knowledge, this is the first study demonstrating the expression of <em>5α</em>-reductases in canine skin and the expression of <em>5α</em>R3 in this tissue. These results may help to elucidate the pathogenesis of alopecia X and to determine more appropriate treatments for this disorder.

Previously, we reported that 6-(3,4-dihydro-1H-isoquinolin-2-yl)-N-(6-methylpyridin-2-yl) nicotinamide (DIMN) analogues inhibited the growth of prostate cancer cells as an anti-androgenic compound. In the present study, we evaluated cytotoxic effects of these DIMN derivatives and found that DIMN-26 most potently inhibited the proliferation of the LNCap-LN3 androgen-dependent and DU145 androgen-independent prostate cancer cells through induction of G2/M phase cell cycle arrest and subsequent apoptosis. The G2/M phase arrest was found due to increases in the activation of cdc2 (also known as cyclin-dependent kinase 1, CDK1)/cyclin B1 complex. DIMN-26 also induced apoptosis in LNCap-LN3 and DU145 prostate cancer cells through activation of caspase-3, -8, and -9, and cleavage of poly(ADP-ribose) polymerase-1 (PARP-1). In addition, DIMN-26 caused the dephosphorylation and mitochondrial accumulation of Bad protein and induced the loss of mitochondria membrane potential, consequently releasing cytochrome c into the cytosol of the cell. Furthermore, overexpression of AKT protein significantly reduced DIMN-26-induced PARP-1 cleavage and p-Bad decrease and cdc2 activation. In addition, DIMN-26 inhibited the <em>5α</em>-dihydrotestosterone (<em>DHT</em>)-induced cell growth and proliferation and nuclear translocation and transcriptional activities of androgen receptor (AR) in LNCap-LN3 prostate cancer cells. Consistent with these findings, DIMN-26 significantly inhibited the <em>DHT</em>-induced expression of AR-response genes (ARGs), such as prostate-specific antigen (PSA), AR, β2-microglobulin (B2M), selenoprotein P (SEPP1), and ste20-related proline-alanine-rich kinase (SPAK) in LNCap-LN3 prostate cancer cells. Taken together, these results suggest that DIMN-26 plays a therapeutic role not only in induction of G2/M arrest and apoptosis but also in suppression of androgen receptor signaling in androgen-dependent and androgen-independent prostate cancer cells.

The <em>5α</em>-reductase converts testosterone to dihydrotestosterone (<em>DHT</em>), and excess <em>DHT</em> could cause androgen-related diseases such as androgenetic alopecia and benign prostatic hyperplasia (BPH). To discover new <em>5α</em>-reductase inhibitors, effective drug screening method with high throughput is thus essential. In this study, fully automated chip-based nanoelectrospray ionization-mass spectrometry (nano-ESI-MS) was innovatively utilized as a screening tool for <em>5α</em>-reductase inhibitory assay in direct infusion mode, which simplified sample pretreatment and greatly improved experimental efficiency. The preliminary data indicated that curcumin, a natural anti-inflammatory compound, exhibited notably <em>5α</em>-reductase inhibition activity. Moreover, the obtained results of the chip-based nano-ESI-MS were well consistent with those of HPLC-MS, which suggested that the chip-based nano-ESI-MS could be treated as a rapid and high-throughput drugs screening strategy in pharmaceutical development. Graphical abstract.

Benign prostatic hyperplasia (BPH) can lead to lower urinary tract symptoms. Rape pollen is an apicultural product that is composed of nutritionally valuable and biologically active substances. The aim of the present study was to investigate the protective effect of rape pollen supercritical CO2 fluid extract (SFE-CO2) in BPH development using a testosterone-induced BPH rat model. BPH was induced in the experimental groups by daily subcutaneous injections of testosterone for a period of 30 days. Rape pollen SFE-CO2 was administered daily by oral gavage concurrently with the testosterone injections. Animals were sacrificed at the scheduled termination and the prostates were weighed and subjected to histopathological examination. Testosterone, dihydrotestosterone (<em>DHT</em>), <em>5α</em>-reductase and cyclooxygenase-2 (COX-2) levels were also measured. BPH-induced animals exhibited an increase in prostate weight with increased testosterone, <em>DHT</em>, <em>5α</em>-reductase and COX-2 expression levels. However, rape pollen SFE-CO2 treatment resulted in significant reductions in the prostate index and testosterone, <em>DHT</em>, <em>5α</em>-reductase and COX-2 levels compared with those in BPH-induced animals. Histopathological examination also demonstrated that rape pollen SFE-CO2 treatment suppressed testosterone-induced BPH. These observations indicate that rape pollen SFE-CO2 inhibits the development of BPH in rats and these effects are closely associated with reductions in <em>DHT</em>, <em>5α</em>-reductase and COX-2 levels. Therefore, the results of the present study clearly indicate that rape pollen SFE-CO2 extract may be a useful agent in BPH treatment.

The aim of the present study was to examine the effect of prolonged use of finasteride on serum levels of dihydrotestosterone (<em>DHT</em>), estradiol (E2), progesterone, testosterone and androstenedione in women during the menstrual period. Further, to screen and compare the <em>5α</em>-reductase activities through the expression of SRD5A1, SRD5A2 and AR gene and to determine the level of VEGF, VKOR and SAA gene expression and DNA damage. A total of 30 Saudi women aged between 25 to 35 years were enrolled in the study. The selected women were divided into two groups. The first group (n=15) received 5 mg finasteride/day for prolonged period of one year and second group (n=15) was taken as a healthy control. ELISA technique was used for measuring the serum levels of the targeted hormones and Comet assay was used for checking the DNA integrity.Our findings revealed significant decrement of <em>DHT</em>, E2, p sterone and androstenedione levels and elevated levels of testosterone in group treated with daily oral doses of 5 mg finasteride/day compared to the control subjects. mRNA expression suggested that finasteride has concrete effects on the gene expression of the selected genes from the treated group in comparison to the control group. In addition, finasteride induced DNA damage and heavy menstrual bleeding was noted in women treated with finasteride. In conclusion, the present findings revealed that finasteride has adverse health effects in women associated with gonadal sex steroids alterations, DNA damage and heavy menstrual bleeding with no consensus in treatment of androgenetic alopecia in women.

Androgen receptor (AR) signaling is essential for prostate cancer (PC) progression and treatment. Experiments have demonstrated that the intratumoral androgen levels are not affected by circulating androgen levels, but rather modulated by local steroid-converting enzyme activities. The expression modulation status of human steroid-converting enzymes and nuclear receptors are of great promise to identify novel therapeutic targets. Meta-analysis was performed with 9 cohorts (1093 specimens) from Gene Expression Omnibus, 16 cohorts (933 specimens) from Oncomine and the TCGA cohort (550 specimens). We found significant up regulation of <em>5α</em>-reductase type 1 and type 3 in both primary and metastatic PC, together with the down regulation of AKR1C2 in primary PC, contributing to the high intratumoral <em>DHT</em> levels. The expression of AR in metastatic PC was up regulated, indicating the importance of AR signaling in the progression of this cancer. The down regulations of HSD11B1 and NR3C1 in primary and metastatic PC may diminish the anti-inflammation and anti-proliferation effects of glucocorticoids signaling. Furthermore, the decrease of progesterone receptor (PGR) expression in primary and metastatic PC was also observed, relieving the suppression effect of PGR on PC proliferation. The clinical evidences of the remarkable expression modulation of steroid-converting enzymes and receptors in PC may indicate novel combined treatment against this highly incident cancer.

Because human prostate-distributed UDP-glucuronosyltransferase (UGT) 2B15 metabolizes <em>5α</em>-dihydrotestosterone (<em>DHT</em>) and 3α-androstane-<em>5α</em>,17β-diol metabolite, we sought to determine whether 2B15 requires regulated phosphorylation similar to UGTs already analyzed. Reversible down-regulation of 2B15-transfected COS-1 cells following curcumin treatment and irreversible inhibition by calphostin C, bisindolylmaleimide, or röttlerin treatment versus activation by phorbol 12-myristate 13-acetate indicated that 2B15 undergoes PKC phosphorylation. Mutation of three predicted PKC and two tyrosine kinase sites in 2B15 caused 70-100 and 80-90% inactivation, respectively. Anti-UGT-1168 antibody trapped 2B15-His-containing co-immunoprecipitates of PKCα in 130-140- and >150-kDa complexes by gradient SDS-PAGE analysis. Complexes bound to WT 2B15-His remained intact during electrophoresis, whereas 2B15-His mutants at phosphorylation sites differentially dissociated. PKCα siRNA treatment inactivated >50% of COS-1 cell-expressed 2B15. In contrast, treatment of 2B15-transfected COS-1 cells with the Src-specific activator 1,25-dihydroxyvitamin D(3) enhanced activity; treatment with the Src-specific PP2 inhibitor or Src siRNA inhibited >50% of the activity. Solubilized 2B15-His-transfected Src-free fibroblasts subjected to in vitro [γ-(33)P]ATP-dependent phosphorylation by PKCα and/or Src, affinity purification, and SDS gel analysis revealed 2-fold more radiolabeling of 55-58-kDa 2B15-His by PKCα than by Src; labeling was additive for combined kinases. Collectively, the evidence indicates that 2B15 requires regulated phosphorylation by both PKCα and Src, which is consistent with the complexity of synthesis and metabolism of its major substrate, <em>DHT</em>. Whether basal cells import or synthesize testosterone for transport to luminal cells for reduction to <em>DHT</em> by <em>5α</em>-steroid reductase 2, comparatively low-activity luminal cell 2B15 undergoes a complex pattern of regulated phosphorylation necessary to maintain homeostatic <em>DHT</em> levels to support occupation of the androgen receptor for prostate-specific functions.

While androgen binding sites have been localized to motoneurons of the lateral motor column (LMC) of 10-18-day chick embryo spinal cord, they have yet to be characterized biochemically. Studies were undertaken to characterize the binding of the androgen [3H]<em>5a</em>-dihydrotestosterone ([3H]<em>DHT</em>) to spinal cord cytosols. Saturable, high-affinity binding of [3H]<em>DHT</em> to cytosols prepared from both 6 and 10-day spinal cords was observed. The binding component was a macromolecular species as it displayed a sedimentation coefficient of 8S upon centrifugation in sucrose gradients; proteinaceous, as binding was eliminated by heating cytosols; and displayed steroid-specific binding, as other non-androgen steroid agonists did not significantly inhibit [3H]<em>DHT</em> binding. The number of binding sites increased 10-fold from embryonic day 6 to day 10. The availability of testosterone and presence of androgen receptors in spinal cord motoneurons as spontaneous limb motility begins suggests a possible role for androgen-receptor-dependent gene expression in the process of target-dependent differentiation of LMC motoneurons.

In this study, proton affinitive derivatization using picolinic acid and its analogs (3- and 6-methylpicolinic acid and 5-butylpicolinic acid) with proton affinitive moieties was performed for the highly sensitive determination of testosterone (T) and <em>5α</em>-dihydrotestosterone (<em>DHT</em>) in saliva by LC-ESI-MS/MS. T and <em>DHT</em> were converted to their corresponding picolinate esters and their chromatographic behavior was investigated with a reversed phase column. The picolinate ester of each steroid exhibited a clear single peak and elution occurred in the following order: picolinate, 3/6-methylpicolinate, and 5-butylpicolinate. Estimation and understanding of the separation and retention time of each picolinate ester was made simple using the develop method. Although the peaks of picolinate and 3/6-methylpicolinate esters were suppressed by interference from the saliva background (matrix effect), the 5-butylpicolinate esters were only marginally affected.

This article provides data in support of the research article entitled "Rapid uptake, biotransformation, esterification and lack of depuration of testosterone and its metabolites by the common mussel, Mytilus spp." (T.I. Schwarz, I. Katsiadaki, B.H. Maskrey, A.P. Scott, 2017) [1]. The uptake of tritiated testosterone (T) from water by mussels is presented. The two main radioactive peaks formed from T and present in the fatty acid ester fraction of mussel tissues were shown to have the same elution positions on a thin layer chromatography plate as 17β-hydroxy-<em>5α</em>-androstan-3-one (<em>DHT</em>) and <em>5α</em>-androstan-3β,17β-diol (3β,17β-A<em>5α</em>). Reverse phase high performance liquid chromatography of the non-esterified (80% ethanol) fraction of the mussel tissue extracts also presented radioactive peaks at the elution positions of <em>DHT</em> and 3β,17β-A<em>5α</em>. There was no evidence for sulfated T in this fraction. It was shown that aeration led to significant losses of radiolabeled testosterone from the water column.

Mussels (Mytilus galloprovincialis) were exposed to different concentrations of testosterone (T: 20, 200 and 2000ng/L) in a semi-static water regime (1-day dosing intervals) for up to 5 days in an attempt to see whether endogenous steroid levels and steroid metabolism were altered by exogenous exposure to testosterone. Whole tissue levels of total testosterone (free+esterified) sharply increased in a concentration-dependent manner, from 2ng/g in controls to 290ng/g in organisms exposed to the highest concentration. In contrast, levels of free testosterone were only significantly elevated at the high-exposure group (5-fold increase with respect to controls). Increased activity of palmitoyl-CoA:testosterone acyltransferase (ATAT) was detected in organisms exposed to the highest concentration of testosterone, while those exposed to low and medium concentrations showed significant alterations in their polyunsaturated fatty acid profiles. The obtained results suggest that esterification of the excess of T with fatty acids might act as a homeostatic mechanism to maintain endogenous levels of free T stable. Interestingly, a decrease in CYP3A-like activity was detected in T-exposed mussels together with a significant decrease in the metabolism of the androgen precursor androstenedione to dihydrotestosterone (<em>5α</em>-<em>DHT</em>). Overall, the work contributes to the better knowledge of androgen metabolism in mussels.

A series of experiments were conducted to investigate the effect of rutting season on metabolism of testosterone (T) and its effect on drug metabolizing enzymes in dromedary camels. Serum and tissue samples were collected from liver, testes and poll glands of rutting and non- rutting camels treated with T at a dose of 0.5 mg/kg or <em>5α</em>-dihydrotestosterone (<em>DHT</em>) at a dose of 0.2 mg/kg, given intramuscularly for 7 days. Liver samples were also used to monitor drug metabolizing enzymes. Testosterone and <em>DHT</em> concentrations were significantly (<i>P</i><0.05) increased in testicular tissue and peripheral circulation of rutting camels compared to non-rutting camels and in non-rutting camels treated with T or <em>DHT</em>. Drug metabolizing enzymes of phase-1 reaction were significantly (<i>P</i><0.05) inhibited in livers of rutting camels and in non-rutting camels treated with T and <em>DHT</em>. It is suggested that co-administration of drugs metabolized by oxidation with androgens should be avoided. Such drugs may cause adverse drug reaction in rutting camels.

Keywords: androgens; drug metabolizing enzymes; poll gland; rutting camel.

Female Thamnophis sirtalis were administered intraperitoneal implants of either estradiol 17β (E2), testosterone (T), <em>5α</em>-dihydrotestosterone (<em>DHT</em>), or empty silastic capsules for 3 weeks. Plasma levels of E2 and T, measured by specific radioimmunoassay, were significantly elevated in E2 and T-implanted females when compared to controls. T-implanted females did not have elevated circulating E2 levels, suggesting that E2 in the plasma normally is not derived from peripheral conversion of T to E2. Implantation of <em>DHT</em> did not significantly change circulating levels of E2, T, or <em>DHT</em>. All three sex steroid-treated groups of animals had increased oviductal mass compared to controls, while hepatic mass of only E2-treated animals was significantly greater. None of the steroid treatments influenced ovarian mass. Oviductal epithelial cell height and area were greater in the three steroid-treated groups. Testosterone increased myometrial area while <em>DHT</em> drastically altered oviductal morphology. Hepatic cell area and number increased significantly in E2-treated females. However, a small increase in both hepatic cell area and number was noted in T- and <em>DHT</em>-treated females as well. These results suggest that androgen in both an aromatizable and non-aromatizable form can affect the oviduct of females but that the liver primarily responds to estrogenic steroids. © 1992 Wiley-Liss, Inc.

Castrate-resistant prostate cancer (CRPC) is a fatal, metastatic form of prostate cancer, characterized by reactivation of the androgen axis. Aldo-keto reductase 1C3 (AKR1C3) converts androstenedione (AD) and <em>5α</em>-androstanedione to testosterone (T) and <em>5α</em>-dihydrotestosterone (<em>DHT</em>), respectively. In CRPC, AKR1C3 is upregulated and implicated in drug resistance, and has been regarded as a potential therapeutic target. Here we examined a series of indole derivatives containing benzoic acid or phenylhydroxamic acid and found that 4-((3-((3,4,5-trimethoxyphenyl)thio)-1 H -indol-1-yl)methyl)benzoic acid ( 3e ) and N-hydroxy-4-((3-((3,4,5-trimethoxyphenyl)thio)-1 H -indol-1-yl)methyl)benzamide ( 3q ) inhibited 22Rv1 cells proliferation with IC 50 value of 6.37 μm and 2.72 μm, respectively. In enzymatic assay, compounds 3e and 3q exhibited potent inhibitory effect against AKR1C3 (IC 50 = 0.26 and 2.39 μm, respectively). These results indicated that compounds 3e and 3q might be useful leads for further investigation of more potential AKR1C3 inhibitors used for CRPC.

Keywords: AKR1C3; CRPC; Indole; benzoic acid; cytotoxicity.

In this study, we show that during normal rat pregnancy, there is a gestational stage-dependent decrease in androgen receptor (AR) abundance in the gravid uterus and that this is correlated with the differential expression of endometrial receptivity and decidualization genes during early and mid-gestation. In contrast, exposure to <em>5α</em>-dihydrotestosterone (<em>DHT</em>) and insulin (INS) or <em>DHT</em> alone significantly increased AR protein levels in the uterus in association with the aberrant expression of endometrial receptivity and decidualization genes, as well as disrupted implantation. Next, we assessed the functional relevance of the androgen-AR axis in the uterus for reproductive outcomes by treating normal pregnant rats and pregnant rats exposed to <em>DHT</em> and INS with the anti-androgen flutamide. We found that AR blockage using flutamide largely attenuated the <em>DHT</em> and INS-induced maternal endocrine, metabolic, and fertility impairments in pregnant rats in association with suppressed induction of uterine AR protein abundance and androgen-regulated response protein and normalized expression of several endometrial receptivity and decidualization genes. Further, blockade of AR normalized the expression of the mitochondrial biogenesis marker Nrf1 and the mitochondrial functional proteins Complexes I and II, VDAC, and PHB1. However, flutamide treatment did not rescue the compromised mitochondrial structure resulting from co-exposure to <em>DHT</em> and INS. These results demonstrate that functional AR protein is an important factor for gravid uterine function. Impairments in the uterine androgen-AR axis are accompanied by decreased endometrial receptivity, decidualization, and mitochondrial dysfunction, which might contribute to abnormal implantation in pregnant PCOS patients with compromised pregnancy outcomes and subfertility. KEY MESSAGES: The proper regulation of uterine androgen receptor (AR) contributes to a normal pregnancy process, whereas the aberrant regulation of uterine AR might be linked to polycystic ovary syndrome (PCOS)-induced pregnancy-related complications. In the current study, we found that during normal rat pregnancy there is a stage-dependent decrease in AR abundance in the gravid uterus and that this is correlated with the differential expression of the endometrial receptivity and decidualization genes Spp1, Prl, Igfbp1, and Hbegf. Pregnant rats exposed to <em>5α</em>-dihydrotestosterone (<em>DHT</em>) and insulin (INS) or to <em>DHT</em> alone show elevated uterine AR protein abundance and implantation failure related to the aberrant expression of genes involved in endometrial receptivity and decidualization in early to mid-gestation. Treatment with the anti-androgen flutamide, starting from pre-implantation, effectively prevents <em>DHT</em> + INS-induced defects in endometrial receptivity and decidualization gene expression, restores uterine mitochondrial homeostasis, and increases the pregnancy rate and the numbers of viable fetuses. This study adds to our understanding of the mechanisms underlying poor pregnancy outcomes in PCOS patients and the possible therapeutic use of anti-androgens, including flutamide, after spontaneous conception.

Keywords: Androgen receptor; Flutamide; Implantation; Mitochondrial function; Polycystic ovary syndrome; Pregnant uterus.

The development and maintenance of prostate function depend on a fine balance between oestrogen and androgen levels. Finasteride inhibits <em>5α</em>-reductase, which is responsible for the conversion of testosterone into its most active form, dihydrotestosterone (<em>DHT</em>). Enzymes that metabolize these hormones have a highly relevant role in both the normal prostate metabolism and in the occurrence of pathological conditions. There are few studies on the impact of finasteride on male prostate development and fewer studies on the female prostate and possible intersexual differences. Therefore, we treated male and female gerbils from 7-14 days in postnatal life with a high dose of finasteride (500 μg/Kg/d); the prostate complexes were then removed and submitted to immunohistochemistry, immunofluorescence and three-dimensional reconstruction. In addition, hormonal serum dosages were administered. Treatment with finasteride resulted in an increased thickness of the periductal smooth musculature in the prostate of both male and female gerbils, such as well as a reduction in the thickness of developing prostate alveoli in both sexes. In addition, intersexual differences were observed as increased epithelial proliferation and decreases in the number of developing alveoli in females. Together, the data indicate that postnatal exposure to finasteride causes greater changes in the female gerbil prostate than in the male. This article is protected by copyright. All rights reserved.

<AbstractText><em>5α</em>-reductase type 2 deficiency (<em>5α</em>RD) is an autosomal recessive hereditary disease of the group of 46, XY disorders of sex development (DSD).</AbstractText><AbstractText>To study the growth pattern in Chinese pediatric patients with <em>5α</em>RD.</AbstractText><AbstractText>Data were obtained from 141 patients with <em>5α</em>RD (age: 0-16 years old) who visited eight pediatric endocrine centers from January 2010 to December 2017.</AbstractText><AbstractText>In this retrospective cohort study, height, weight, and other relevant data were collected from the multicenter hospital registration database. Baseline luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), and dihydrotestosterone (<em>DHT</em>) after human chorionic gonadotropin (HCG) stimulation test were measured by enzyme enhanced chemiluminescence assay. Bone age (BA) was assessed using the Greulich-Pyle (G-P) atlas. Growth curve was constructed based on λ-median-coefficient of variation method (LMS).</AbstractText><p><div><b>Results</b></div>The height standard deviation scores (HtSDS) and weight standard deviation scores (WtSDS) in <em>5α</em>RD children were in the normal range as compared to normal boys. Significantly higher HtSDS was observed in patients with <em>5α</em>RD who were <1 year old (<i>t</i> = 3.658, 2.103, <i>P</i> = 0.002, 0.048, respectively), and higher WtSDS in those <6 months old (<i>t</i> = 2.756, <i>P</i> = 0.012). Then HtSDS and WtSDS decreased gradually and fluctuated near the median of the same age until 13 years. WtSDS in <em>5α</em>RD children from northern China were significantly higher than those from the south (<i>Z</i> = -2.670, <i>P</i> = 0.008). The variation tendency of HtSDS in Chinese <em>5α</em>RDs was consistent with the trend of stimulating T. HtSDS and stimulating T in the external masculinization score (EMS) <7 group were slightly higher than those in EMS ≥ 7 group without significant difference. Additionally, the ratio of BA over chronological age (BA/CA) was significantly <1 in children with <em>5α</em>RD.</p><AbstractText>Children with <em>5α</em>RD had a special growth pattern that was affected by high levels of T, while <em>DHT</em> played a very small role in it. Their growth accelerated at age <1 year, followed by slowing growth and fluctuating height near normal median boys' height. The BA was delayed in <em>5α</em>RD children. Androgen treatment, which may be considered anyway for male <em>5α</em>RD patients with a micropenis, may also be beneficial for growth.</AbstractText>

<AbstractText>Taxifolin, a flavonoid, has several pharmacological activities. Taxifolin inhibits some steroid biosynthetic enzymes, suggesting that it might inhibit neurosteroid synthesis. Here, we investigated effects of taxifolin on rat neurosteroidogenic enzymes, steroid <em>5α</em>-reductase 1 -(SRD5A1), 3α-hydroxysteroid dehydrogenase (AKR1C9), and retinol dehydrogenase 2 (RDH2).</AbstractText><AbstractText>SRD5A1, AKR1C9, and RDH2 were expressed in COS-1 cells and the direct effects of taxifolin on these enzymes and the mode of action were determined using steroid substrates and thin chromatography separation-coupled radiometric scanning.</AbstractText><AbstractText>The half maximal inhibitory concentration values of taxifolin on SRD5A1 and AKR1C9 were 100 and 1.01 mol/L. Taxifolin did not inhibit RDH2 even at as high as 100 mol/L. Taxifolin competitively inhibited rat AKR1C9 against steroid dihydrotestosterone (<em>DHT</em>), and docking analysis revealed that taxifolin bound to the steroid binding site of rat AKR1C9 with similar free energy with the substrate <em>DHT</em>.</AbstractText><AbstractText>Taxifolin is a potent inhibitor of rat AKR1C9 and moderate inhibitor of rat SRD5A1, thereby controlling the rate of neurosteroid biosynthesis.</AbstractText>

BACKGROUND

Pituitary luteinizing hormone (LH) stimulates testicular production of testosterone (T) which is metabolized to dihydrotestosterone (<em>DHT</em>) by <em>5α</em>-reductase and to oestradiol (E2) by aromatase. How the activity of population variants in these enzymes impacts on gonadal function is unclear. We examined whether polymorphisms in <em>5α</em>-reductase (SRD5A2) and aromatase (CYP19A1) genes predict circulating sex hormone concentrations.

METHODS

Cross-sectional analysis of 1865 community-dwelling men aged 50.4 ± 16.8 years.

METHODS

Early morning sera assayed for T, DHT and E2 (mass spectrometry), and SHBG and LH (immunoassay). Two SRD5A2 and eleven CYP19A1 polymorphisms were analysed by PCR. Regression models were adjusted for age and cardiometabolic risk factors.

RESULTS

SRD5A2 polymorphism rs9282858 GA vs. GG was associated with higher serum T (+1.5 nmol/L, P < 0.001) and higher SHBG (+3.3 nmol/L, P = 0.001). CYP19A1 polymorphisms were associated with higher serum E2 and lower LH in reciprocal fashion, from which the two-copy haplotype rs10046 = T/rs2899470 = G/rs11575899 = I/rs700518 = G/rs17703883 = T was associated with higher E2 (63.4 vs. 56.5 pmol/L, P = 0.001) and lower LH (3.9 vs. 4.5 IU/L, P = 0.001) compared to null copies. Conversely, rs10046 = C/rs2899470 = T/rs11575899 = D/rs700518 = A/rs17703883 = C was associated with lower E2 (51.8 vs. 62.0 pmol/L, P = 0.001) and higher LH (5.7 vs. 3.9 IU/L, P < 0.001). These haplotypes were associated primarily with differences in E2 in men <65 years and LH in men ≥65 years.

CONCLUSIONS

A <em>5α</em>-reductase polymorphism predicts circulating T and SHBG, while aromatase polymorphisms predict E2 and LH in reciprocal fashion. Age and aromatase polymorphisms interact to affect E2 and LH. How these functional polymorphisms impact on male reproductive and general health outcomes requires further study.