The steroid hormones not only exert various endocrine functions but also act as the autocrine or paracrine factors in different tissues of mammals. In the present study, the seasonal expressions of androgen receptor (AR), estrogen receptors alpha and beta (ERα and ERβ), aromatase cytochrome P450 (P450arom) and <em>5α</em>-reductase 1, 2 were investigated in the epididymis of the muskrat. HE staining showed enlarged lumen and abundant sperm in the breeding season while reduced lumen with no sperm in the non-breeding season. The staining of AR was presented in nuclei of epithelial cells of the epididymis in both seasons. The immunostaining of ERα was localized in both nuclei and cytoplasm of epithelial cells of the epididymis during the breeding season, while the weak staining of ERα was only in the nuclei of epithelial cells during the non-breeding season. In contrast, ERβ signal was negative in the epididymis of the muskrat in both seasons. The positive signals for P450arom and <em>5α</em>-reductase 1, 2 were found in the cytoplasm of epithelial and smooth muscle cells during both seasons. Moreover, the protein and mRNA expression levels of AR, ERα, P450arom and <em>5α</em>-reductase 1, 2 were significantly higher in the epididymis during the breeding season than those of the non-breeding season, and the expression level of <em>5α</em>-reductase 1 was higher when compared with <em>5α</em>-reductase 2. In addition, the levels of testosterone (T) and dihydrotestosterone (<em>DHT</em>) in the epididymis and serum were remarkably higher during the breeding season. Taken together, these findings suggested androgen and estrogen might play an important endocrine or autocrine/paracrine role to regulate the epididymal functions of the muskrat.

Porcine testicular carbonyl reductase, PTCR which is one of the short chain dehydrogenases/reductases (SDR) superfamily catalyzes the NADPH-dependent reduction of carbonyl compounds including steroids and prostaglandins. Previously we reported C-terminal tail of PTCR was deleted due to a nonsynonymous single nucleotide variation (nsSNV). Here we identified from kinetic studies that the enzymatic properties for <em>5α</em>-dihydrotestosterone (<em>5α</em>-<em>DHT</em>) were different between wild-type and C-terminal-deleted PTCRs. Compared to wild-type PTCR, C-terminal-deleted PTCR has much higher reduction rate. To investigate structural difference between wild-type and C-terminal-deleted PTCRs upon <em>5α</em>-<em>DHT</em> binding, we performed molecular dynamics simulations for two complexes. Using trajectories, molecular interactions including hydrogen bonding patterns, distance between <em>5α</em>-<em>DHT</em> and catalytic Tyr193, and interaction energies are analyzed and compared. During the MD simulation time, the dynamic behavior of C-terminal tail in wild-type PTCR is also examined using essential dynamics analysis. The results of our simulations reveal that the binding conformation of <em>5α</em>-<em>DHT</em> in C-terminal-deleted PTCR is more favorable for reduction reaction in PTCR, which shows strong agreement with kinetic data. These structural findings provide valuable information to understand substrate specificity of PTCR and further kinetic properties of enzymes belonging to the SDR superfamily.

Steroid <em>5α</em>-reductase type 2 deficiency (<em>5α</em>RD2) is a congenital disorder of sex development caused by impairment of conversion from testosterone (T) to <em>5α</em>-dihydrotestosterone (<em>DHT</em>). <em>DHT</em> deficiency leads to various degrees of undervirilized external genitalia including micropenis, primarily correlated with mutations of the SRD5A2 gene that encodes <em>5α</em>-reductase type 2. Four Japanese boys with isolated micropenis were diagnosed as <em>5α</em>RD2 by elevated ratios of serum T/<em>DHT</em>, and decreased ratios of urinary <em>5α</em>/5β-reduced steroid metabolites. Genetic analyses for SRD5A2 identified that the four patients shared a hypomorphic mutation R227Q that has a residual activity related to the mild-form of <em>5α</em>RD2. For prepubertal micropenis, <em>DHT</em> was transdermally applied to the four patients at the ages of 4-11 year, increasing a median of stretched penile lengths (SPLs) from 2.6 cm (-2.5 SD) to 4.4 cm (-0.2 SD). Nevertheless, the post-pubertal penile growth was apparently retarded, despite normal levels of T secreted from well-developed testes. The second course of <em>DHT</em> treatment underwent at ages of 12-18 year, but unable to normalize SPLs at a range of 6.0 to 7.0 cm (-3.4 to -2.4 SD). The prostate volumes of two patients were variable at 8.1 and 21 cm³, and a sperm cell count of one patient was normal as young adult. <em>DHT</em> treatment contributes to development of the penis and prostate, which are favorable for the potential fertility of <em>5α</em>RD2 adults. Meanwhile, the retarded penile growth and a risk of prostate overgrowth may complicate the post-pubertal management with <em>DHT</em> for <em>5α</em>RD2 males.

Disorders of sex development (DSD) are a wide-ranging group of complex conditions that influence chromosomal, gonadal, and phenotypic sex. The prevalence of DSD is very low, but affected patients deserve individualized management to improve psychological, sexual, and reproductive outcomes. This review aims to clarify the fertility potential of DSD patients who can be reared as females and their chance of becoming pregnant, especially using assisted reproductive techniques (ART). Due to the effects of DSD on internal and external genital organs, these conditions result in varying degrees of fertility potential. Fertility rate depends on the phenotype and is inversely related to the severity of the disorder. Reproductive endocrinologists and infertility specialists must be considered active partners of the interdisciplinary treatment team. With current advances in ART, pregnancy is more achievable in patients who were considered infertile at first glance. Due to the complexity of the medical management in DSD patients, more studies should be conducted to conclusively suggest the best choice for improving their fertility potential.<b>Abbreviations</b>: AIS: Androgen Insensitivity Syndrome; AMH: Anti-Müllerian Hormone; ART: Assisted Reproductive Technology; ASRM: American Society for Reproductive Medicine; CAH: Congenital Adrenal Hyperplasia; CAIS: Complete Androgen Insensitivity Syndrome; <em>DHT</em>: Dihydrotestosterone; DSD: Disorders of Sexual Development; FSH: Follicle Stimulating Hormone; GD: Gonadal Dysgenesis; ICSI: Intracytoplasmic Sperm Injection; IUGR: Intrauterine Growth Restriction; IVF: In Vitro Fertilization; IVF-ET: IVF and Embryo Transfer; LH: Luteinizing Hormone; MGD: Mixed Gonadal Dysgenesis; MRI: Magnetic Resonance Imaging; MRKH: Mayer-Rokitansky-Kuster-Hauser; US: Ultrasonography; HSG: Hysterosalpingography; PAIS: Partial Androgen Insensitivity Syndrome; PGD: Preimplantation Genetic Diagnosis; POR: P450 Oxidoreductase; PROM: Premature Rupture of Membranes; TS: Turner Syndrome; 17β-HSD III: 17β-Hydroxysteroid Dehydrogenase III; 21-OHD: 21-hydroxylase deficiency; <em>5α</em>-RD-2: <em>5α</em>-reductase-2.

Keywords: Disorders of sex development; female fertility; reproductive techniques.

<strong class="sub-title"> Background: </strong> A growing body of data indicates that the physiology of the liver is sex-hormone dependent, with some types of liver failure occurring more frequently in males, and some in females. In males, in physiological conditions, testosterone acts via androgen receptors (AR) to increase insulin receptor (IR) expression and glycogen synthesis, and to decrease glucose uptake controlled by liver-specific glucose transporter 2 (GLUT-2). Our previous study indicated that this mechanism may be impaired by finasteride, a popular drug used in urology and dermatology, inhibiting <em>5α</em>-reductase 2, which converts testosterone (T) into dihydrotestosterone (<em>DHT</em>). Our research has also shown that the offspring of rats exposed to finasteride have an altered T-<em>DHT</em> ratio and show changes in their testes and epididymides. Therefore, the goal of this study was to assess whether the administration of finasteride had an trans-generational effect on (i) GLUT-2 dependent accumulation of glycogen in the liver, (ii) IR and AR expression in the hepatocytes of male rat offspring, (iii) a relation between serum T and <em>DHT</em> levels and the expression of GLUT2, IR, and AR mRNAs, (iv) a serum glucose level and it correlation with GLUT-2 mRNA.

Methods: The study was conducted on the liver (an androgen-dependent organ) from 7, 14, 21, 28, and 90-day old Wistar male rats (F1:Fin) born by females fertilized by finasteride-treated rats. The control group was the offspring (F1:Control) of untreated Wistar parents. In the histological sections of liver the Periodic Acid Schiff (PAS) staining (to visualize glycogen) and IHC (to detect GLUT-2, IR, and AR) were performed. The liver homogenates were used in qRT-PCR to assess GLUT2, IR, and AR mRNA expression. The percentage of PAS-positive glycogen areas were correlated with the immunoexpression of GLUT-2, serum levels of T and DHT were correlated with GLUT-2, IR, and AR transcript levels, and serum glucose concentration was correlated with the age of animals and with the GLUT-2 mRNA by Spearman's rank correlation coefficients.

Results: In each age group of F1:Fin rats, the accumulation of glycogen was elevated but did not correlate with changes in GLUT-2 expression. The levels of GLUT-2, IR, and AR transcripts and their immunoreactivity statistically significantly decreased in F1:Fin animals. In F1:Fin rats the serum levels of T and DHT negatively correlated with androgen receptor mRNA. The animals from F1:Fin group have statistically elevated level of glucose. Additionally, in adult F1:Fin rats, steatosis was observed in the liver (see Appendix A).

Conclusions: It seems that treating male adult rats with finasteride causes changes in the carbohydrate metabolism in the liver of their offspring. This can lead to improper hepatic energy homeostasis or even hyperglycaemia, insulin resistance, as well as some symptoms of metabolic syndrome and liver steatosis.

Keywords: AR; DHT deficiency; GLUT-2; IR; finasteride; glycogen storage; serum androgens and glucose concentration; zones of hepatic lobules.

Steroid <em>5α</em>-reductase type 2 deficiency (<em>5α</em>-RD2) and androgen insensitivity syndrome (AIS) are difficult to distinguish clinically and biochemically, and adrenal-derived androgens have not been investigated in these conditions using modern methods. The objective of the study was to compare Chinese patients with <em>5α</em>-RD2, AIS, and healthy men. Sixteen patients with <em>5α</em>-RD2, 10 patients with AIS, and 39 healthy men were included. Serum androgen profiles were compared in these subjects using liquid chromatography/tandem mass spectrometry (LC-MS/MS). Based on clinical features and laboratory tests, <em>5α</em>-RD2 and AIS were diagnosed and confirmed by genotyping. Dihydrotestosterone (<em>DHT</em>) and testosterone (T) were both significantly lower in patients with <em>5α</em>-RD2 than AIS (p < 0.0001). The T/<em>DHT</em> ratio was higher in <em>5α</em>-RD2 (4.5-88.6) than AIS (13.4-26.7) or healthy men (7.6-40.5). Using LC-MS/MS, a cutoff T/<em>DHT</em> value of 27.3 correctly diagnosed <em>5α</em>-RD2 versus AIS with sensitivity 93.8% and specificity 100%. Among the adrenal-derived 11-oxygenated androgens, 11β-hydroxyandrostenedione (11OHA4) and 11-ketoandrostenedione (11KA4) were also lower in patients with <em>5α</em>-RD2 than those of patients with AIS. In contrast, 11β-hydroxytestosterone (11OHT) was higher in <em>5α</em>-RD2 than AIS. Furthermore, a 11OHT/11OHA4 cutoff value of 0.048 could also distinguish <em>5α</em>-RD2 from AIS. Thus, both elevated T/<em>DHT</em> values above 27.3 and the unexpected 11-oxygenated androgen profile, with a 11OHT/11OHA4 ratio greater than 0.048, distinguished <em>5α</em>-RD2 from AIS. These data suggest that the metabolism of both gonadal and adrenal-derived androgens is altered in <em>5α</em>-RD2.

Steroid hormones play an essential role in regulating physiological and reproductive development throughout the lifetime of an individual. One of the difficulties in determining endogenous substances is the lack of a blank matrix. Especially when the level of analytes is lower than the level in the so-called blank matrix. In the present study, an optimized HPLC-MS/MS method was developed and validated to quantify androstenedione (ASD), testosterone (Ts), dehydroepiandrosterone (DHEA), <em>5α</em>-dihydrotestosterone (<em>DHT</em>), and progesterone (P) in serum samples from healthy people using PBS (pH = 7.4) as the blank surrogate matrix. Simultaneously, the method investigated the characteristics of NaCl, bull serum albumin, pure water as surrogate matrices for the analysis of steroid hormones. The data showed that the matrix effects of ASD, Ts, DHEA, <em>DHT</em>, and P in the same groups were not significantly different between PBS and twice charcoal-stripped serum (CS₂S) as a blank surrogate matrix. Furthermore, the LLOQ using PBS as the blank matrix was up to 0.005 ng/mL for ASD, Ts, and P and 0.05 ng/mL for DHEA and <em>DHT</em>. The reference ranges of concentration (C_PBS) of 5 steroid hormones were provided. Compared to the concentration with CS₂S (C_CSS) as the blank surrogate matrix, the relative biases (RBs) of Ts, <em>DHT</em>, P, and DHEA were finally stabilized at approximately -0.7%, -15%, -1.2%, and 9.2%, respectively. The results suggest that, in the cases of special required, the developed HPLC-MS/MS method can be used to determine the absolute concentration of 5 hormones in biological samples with PBS as the blank surrogate matrix.

Keywords: HPLC-MS/MS; Human serum; PBS; Quantitation; Steroid hormones; Surrogate matrix.

Background: Spaceflights-induced microgravity can alter various physiological processes in human's body including the functional status of the reproductive system. Rodent model of tail-suspension hindlimb unloading is extensively used to stimulate the organs responses to the microgravity condition. This study explores the potential effects of hindlimb unloading on testicular functions and spermatogenesis in adult male rats and the underlying mechanism/s.

Methods: Twenty Sprague-Dawley rats were allotted into two groups: normally loaded group (control; all arms were in touch with the grid floor) and hindlimb unloaded group (HU; only the forearms were in contact with the grid floor).

<strong class="sub-title"> Results: </strong> Following 30 days of exposure, the HU group saw a decline in body weight, testicular and epidydimal weights, and all semen parameters. The circulating concentrations of gonadotropin-releasing hormone (GnRH), follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone significantly decreased, while levels of kisspeptin, corticosterone, inhibin, prolactin and estradiol (E2) increased in the HU group. Intratesticular levels of <em>5α</em>-reductase enzyme and dihydrotestosterone (<em>DHT</em>) were suppressed, while the levels of aromatase and kisspeptin were significantly elevated in the HU group. Hypothalamic kisspeptin (Kiss1) mRNA expression levels were downregulated while its receptors (Kiss1R) were upregulated in the HU group. On the contrary, the mRNA expression levels of testicular Kiss1 were upregulated while Kiss1R were downregulated. The pituitary mRNA expression levels of FSHβ and LHβ decreased in the HU group. The levels of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and nitric oxide (NO) concentrations, and total antioxidant capacity (TAC) were elevated while malondialdehyde (MDA) concentrations declined in the testes of HU group. The testes of the HU rats showed positive immunostaining of caspase-3, heat shock protein 70 (HSP70) and Bcl2.

Conclusions: Altogether, these results revealed an inhibitory effect of hindlimb unloading on kisspeptin signaling in the hypothalamic-pituitary-testicular axis with impaired spermatogenesis and steroidogenesis.

Keywords: Heat shock protein; Kisspeptin; Microgravity; Testes; Testosterone.

Testosterone (T) plays a crucial role in prostate growth. In androgen-dependent tissues T is reduced to dihydrotestosterone (<em>DHT</em>) because of the presence of the <em>5α</em>-reductase enzyme. This androgen is more active than T, since it has a higher affinity for the androgen receptor (AR). When this mechanism is altered, androgen-dependent diseases, including prostate cancer, could result. The aim of this study was to synthesize several 16-dehydropregnenolone acetate derivatives containing a triazole ring at C-21 and a linear or alicyclic ester moiety at C-3 of the steroidal skeleton. These steroids were designed as potential inhibitors of the activity of both types (1 and 2) of <em>5α</em>-reductase. The cytotoxic activity of these compounds was also evaluated on a panel of PC-3, MCF7, and SK-LU-1 human cancer cell lines. The results from this study showed that with the exception of steroids 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-propionate and 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-pentanoate, the compounds exhibit a lower inhibitory activity for both isoenzymes of <em>5α</em>-reductase than finasteride. Furthermore the 3β-hydroxy-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-20-one and 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-acetate derivatives display 80% cytotoxic activity on the SK-LU-1 cell line. These results also indicated that the triazole derivatives, which have a hydroxyl or acetoxy group at C-3, could have an anticancer effect, whereas the derivatives with a alicyclic ester group at C-3 do not show biological activity.

<b>Background:</b> Graves' disease (GD) is an autoimmune disease that affects more women than men. In our previous study, a potent bioactive androgen, <em>5α</em>-dihydrotestosterone (<em>DHT</em>) showed a protective effect against GD in female BALB/c mice. Evidence indicates that abnormal oxidative stress and immunosuppressive cytokines (TGF-β, IL-35) play critical roles in the pathogenesis and development of GD. The purpose of this research is to measure these cytokines and oxidative stress markers to explore potential protective mechanisms of <em>DHT</em> in a BALB/c mouse model of GD. <b>Methods:</b> GD was induced in female BALB/c mice by intramuscular injection of an adenovirus expressing the A-subunit of the TSH receptor (Ad-TSHR289). <em>DHT</em> or a matching placebo was injected every 3 days. Mice were sacrificed four weeks after the third virus immunization to obtain blood, thyroid and spleen for further analysis. <b>Results:</b> Thyroid hormones were significantly reduced in <em>DHT</em> treated GD mice. In addition, <em>DHT</em> attenuated thyroid oxidative injuries in GD mice, as shown by decreased total antioxidation capability (TAOC), superoxide dismutase (SOD) and the level of malondialdehyde (MDA). The levels of immunosuppressive cytokines (TGF-β, IL-35) in <em>DHT</em> group were significant higher compared with the GD group. <b>Conclusions:</b> The results demonstrated that <em>DHT</em> could reduce the severity of GD in female BALB/c mice by regulating oxidative stress. The upregulation of immunosuppressive cytokines might be another important protective mechanism.

Intratumoural dihydrotestosterone (<em>DHT</em>) synthesis could be an explanation for castration resistance in prostate cancer (PC). By using liquid chromatography-mass spectrometry, we evaluated the intratumoral <em>DHT</em> synthesis from <em>5α</em>-androstane-3β,17β-diol (3β-diol), which is inactive androgen metabolized from <em>DHT</em>. 3β-diol had biochemical potential to be converted to <em>DHT</em> via three metabolic pathways and could stimulate PC cell growth. Especially, 3β-diol was not only converted back to upstream androgens such as dehydroepiandrosterone (DHEA) or Δ5-androstenediol but also converted directly to <em>DHT</em> which is the main pathway from 3β-diol to <em>DHT</em>. Abiraterone had a significant influence on the metabolism of DHEA, epiandrosterone and 3β-diol, by the inhibition of the intratumoural 3β-hydroxysteroid dehydrogenase (3β-HSD) activities which is one of key catalysts in androgen metabolic pathway. The direct-conversion of 3β-diol to <em>DHT</em> was catalysed by 3β-HSD and abiraterone could inhibit this activity of 3β-HSD. These results suggest that PC had a mechanism of intratumoural androgen metabolism to return inactive androgen to active androgen and intratumoural <em>DHT</em> synthesis from 3β-diol is important as one of the mechanisms of castration resistance in PC. Additionally, the inhibition of intratumoural 3β-HSD activity could be a new approach to castration-resistant prostate cancer treatment.

<b>Aim:</b> 16β-¹⁸F-fluoro-<em>5α</em>-dihydrotestosterone (¹⁸F-F<em>DHT</em>) is a radiopharmaceutical that has been investigated as a diagnostic agent for the assessment of androgen receptor (AR) density in prostate cancer using positron emission tomography (PET). However, ¹⁸F-F<em>DHT</em> is rapidly metabolized in humans and excreted via the kidneys into the urine, which can compromise the detection of tumor lesions close to the bladder. Enzalutamide is an AR signaling inhibitor currently used in different stages of prostate cancer. Enzalutamide and its primary metabolite N-desmethylenzalutamide have a comparable affinity for the AR as F<em>DHT</em> but are both mainly excreted via the hepatic route. Radiolabeled enzalutamide could thus be a suitable candidate PET tracer for AR imaging. Here we describe the radiolabeling of enzalutamide with fluorine-18. Moreover, the in-vitro and in-vivo behavior of ¹⁸F-enzalutamide was evaluated and compared to the current standard ¹⁸F-F<em>DHT</em>. <b>Methods:</b> ¹⁸F-enzalutamide was obtained by the fluorination of this nitro precursor. In-vitro cellular uptake studies with ¹⁸F-enzalutamide and ¹⁸F-F<em>DHT</em> were performed in LNCaP (AR+) and HEK293 (AR-) cells. Competition assays with both tracers were conducted in the LNCaP (AR+) cell line. In-vivo PET imaging, ex-vivo biodistribution, and metabolite studies with ¹⁸F-enzalutamide and ¹⁸F-F<em>DHT</em> were conducted in athymic nude male mice bearing an LNCaP xenograft in the shoulder. <b>Results:</b> ¹⁸F-enzalutamide was obtained in 1.4±0.9% radiochemical yield with an apparent molar activity of 6.2±10.3 GBq/µmol. ¹⁸F-F<em>DHT</em> was obtained in 1.5±0.8% yield with a molar activity > 25 GBq/µmol. Co-incubation with an excess of <em>DHT</em> or enzalutamide significantly reduced the cellular uptake of ¹⁸F-enzalutamide and ¹⁸F-F<em>DHT</em> to about 50% in AR+ LNCaP cells, but not in AR- HEK293 cells. PET and biodistribution studies in male mice bearing a LnCaP xenograft showed about 3 times higher tumor uptake for ¹⁸F-enzalutamide than for ¹⁸F-F<em>DHT</em>. Sixty minutes after tracer injection, 93% of ¹⁸F-enzalutamide in plasma was still intact, compared to only 3% for ¹⁸F-F<em>DHT</em>. <b>Conclusion:</b> Despite its lower apparent molar activity, ¹⁸F-enzalutamide shows higher tumor uptake and better metabolic stability than ¹⁸F-F<em>DHT</em> and thus seems to have more favorable properties for imaging of AR with PET. However, further evaluation in other oncological animal models and patients is still warranted to confirm these results.

Keywords: Androgen receptors; Animal Imaging; Enzalutamide; Mice; Oncology: Endocrine; PET imaging; Prostate cancer; Radiopharmaceuticals.

Androgens may exert cardiovascular protective actions by regulating the release and function of different vascular factors. In addition, testosterone (TES) and its 5-reduced metabolites, <em>5α</em>- and 5β-dihydrotestosterone (<em>5α</em>- and 5β-<em>DHT</em>) induce vasorelaxant and hypotensive effects. Furthermore, hypertension has been reported to alter the release and function of the neurotransmitters nitric oxide (NO), calcitonin gene-related peptide (CGRP) and noradrenaline (NA). Since the mesenteric arteries possess a dense perivascular innervation and significantly regulate total peripheral vascular resistance, the objective of this study was to analyze the effect of TES, <em>5α</em>- and 5β-<em>DHT</em> on the neurogenic release and vasomotor function of NO, CGRP and NA. For this purpose, the superior mesenteric artery from male spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY) rats was used to analyze: (i) the effect of androgens (10 nM, incubated for 30 min) on the neurogenic release of NO, CGRP and NA and (ii) the vasoconstrictor-response to NA and the vasodilator responses to the NO donor, sodium nitroprusside (SNP) and exogenous CGRP. The results showed that TES, <em>5α</em>- or 5β-<em>DHT</em> did not modify the release of NO, CGRP or NA induced by electrical field stimulation (EFS) in the arteries of SHR; however, in the arteries of WKY rats androgens only caused an increase in EFS-induced NO release. Moreover, TES, and especially 5β-<em>DHT</em>, increased the vasodilator response induced by SNP and CGRP in the arteries of SHR. These findings could be contributing to the hypotensive/antihypertensive efficacy of 5β-<em>DHT</em> previously described in conscious SHR and WKY rats, pointing to 5β- <em>DHT</em> as a potential drug for the treatment of hypertension.

<AbstractText>Lespedeza cuneata G. Don (Fabaceae), has been used as a traditional treatment of various diseases. There is a report L. cuneata effects on hormone replacement therapy for endocrine-related disease. However, studies related to benign prostatic hyperplasia (BPH) have not been investigated.</AbstractText><AbstractText>The effects of L. cuneata aqueous extract (LCW) on testosterone-induced prostatic hyperplasia (TPH) were examined.</AbstractText><AbstractText>Male Wistar rats (10 weeks, 330-350 g) were randomly divided to 6 groups (n = 6): Control group; TPH group (3 mg/kg, s.c, daily); TPH + LCW (25, 50, 100 mg/kg); TPH + Finasteride 10 mg/kg for 6 weeks. At the end of treatment, histological change of prostate, serum dihydrotestosterone (<em>DHT</em>) level, mRNA expression of <em>5α</em>-reductase, inflammatory factors, proliferating cell nuclear antigen (PCNA) and fibroblast growth factor-2 (FGF-2) in prostate were examined. Then, LCW was treated with BPH-1, a human BPH cell line, at 25, 50, 100 μg/mL for 24 h and examine mRNA level of androgen receptor (AR) and prostate-specific antigen (PSA). In addition, the content of vicenin-2 was analyzed.</AbstractText><AbstractText>LCW treatment of TPH inhibited serum <em>DHT</em> levels by 54.5, 51.2 and 54.1% and mRNA expression of <em>5α</em>-reductase were inhibited 54.3, 61.3 and 73.6%, respectively. In addition, mRNA expression of inflammatory factors, PCNA and FGF-2 were decreased in the prostate of rats. Also, LCW attenuated mRNA level of AR and PSA in BPH-1 cell. The content of vicenin-2 in the LCW was analyzed to 0.89 mg/g.</AbstractText><AbstractText>Based on the results, LCW is a potential pharmacological candidate for the treatment of prostatic hyperplasia.</AbstractText>

Testosterone replacement therapy has a growing interest in daily practice; however, debates on its safety for prostate cancer still continue. Dutasteride-a <em>5α</em>-reductase inhibitor-was shown to be effective in preventing prostate cancer. We therefore aimed to evaluate the effect of testosterone replacement therapy and dutasteride treatment on prostate tissue in castrated rats. Rats were randomised in four groups after bilateral orchidectomy as follows: Group I received testosterone + dutasteride, Group II received only testosterone, Group III had no medical treatment, and Group IV was the control group. After 3 months, rats were sacrificed and laboratory and histopathological examinations were performed. In Groups I and II, prostate volume, T and <em>DHT</em> levels were significantly higher compared to Group III and controls. Groups I and II had also significantly greater preneoplastic histopathological signs; however, in intergroup analyses, Group I showed less premalignant changes compared to Group II. We concluded that dutasteride was effective when combined with testosterone therapy in preventing premalignant histopathological changes in prostate tissue. Further evidence is needed to confirm our findings.

AKR1C3 is a critical enzyme for producing testosterone and <em>5α</em>-<em>DHT</em> in the human body. Inhibiting AKR1C3 is a potential target for treating castration-resistant prostate cancer (CRPC). To find AKR1C3 inhibitors with a new molecular skeleton and binding mode, we analyzed the in vitro inhibitory activity of caffeic acid phenethyl ester (CAPE) and eight other phenolic acid analogues towards AKR1C3 and six other human AKR1 enzymes. We analyzed CAPE and octyl gallate interactions with AKR1C3 using X-ray crystallography, which provided a molecular basis for understanding the phenolic acid inhibitory activity and selectivity towards human AKR1s.

Prostate cancer (PCa) is the most commonly diagnosed cancer in men worldwide. It is essentially dependent on potent androgens, such as testosterone (T) and dihydrotestosterone (<em>DHT</em>). The precursors of T and <em>DHT</em>, which includes androstenedione (A4) and dihydroepiandrosterone (DHEA), and also the metabolites of <em>DHT</em>, <em>5α</em>-androstane-3α,17β-diol (3α-Diol) and <em>5α</em>-androstane-3β,17β-diol (3β-Diol) are able to affect the development of PCa. Therefore, it is important to simultaneously determine all these key androgens. This study aims to develop and validate an LC-MS/MS quantification method to simultaneously detect and quantify the six related androgens, including T, <em>DHT</em>, A4, DHEA, 3α-Diol, and 3β-Diol in limited sample volume. The sample preparation involved liquid extraction with methyl tert-butyl ether (MTBE), following by chemical derivatisation with hydroxylamine. The limits of quantitation for T, <em>DHT</em>, A4, and DHEA were 0.05nM and 3α-Diol and 3β-Diol were 0.5nM with S/N ratio of at least 5:1 by using 100μL samples.

CYP17A1-independent intratumoral steroid hormone synthesis is regarded as one possible explanation for resistance to treatment with the CYP17-inhibitor Abiraterone (Abi). The aim of our study was therefore to investigate the steroid metabolism of prostate cancer cells under serum starvation and the effects of Abi treatment. We assessed steroid metabolism in a panel of prostate cancer cells under serum starvation by radioactivity detector-coupled HPLC and HPLC-ESI-ToF-mass spectrometry after treatment with pregnenolone, progesterone and allopregnanolone. We further evaluated the effects of Abi on steroid metabolism of testosterone, dihydrotestosterone (<em>DHT</em>) and dehydroepiandrosterone (DHEA) by enzyme immunoassays (EIAs). Androgen-responsive cell lines metabolized pregnenolone primarily to mitogenic steroid <em>5α</em>-pregnan-3β,6α-diol-20-one under serum starvation. Co-administration of Abi lead to detectable concentrations of the Abi metabolite Δ4-Abi (D4A), known to inhibit enzymes other than CYP17A1 in steroid metabolism. In addition, co-administration of Abi abrogated pregnenolone metabolism and resulted in a CYP17A1-independent significant increase of DHEA (13- to >100-fold) and <em>DHT</em> (2.5-fold) in androgen-responsive cells. Our results demonstrate the CYP17A1-independent formation of <em>5α</em>-pregnan-3β,6α-diol-20-one by androgen-responsive prostate cancer cells under serum starvation and its inhibition by Abi. Its metabolism from pregnenolone suggests a major steroidogenesis shift in these cells, hinting at a neuroendocrine transdifferentiation phenomenon. The marked increase of DHEA levels by Abi resembles the steroidogenic pathways in nervous tissue, in a manner that precludes CYP17A1 activity. To which extent these processes are responsible or involved in the development of resistance to Abi, needs to be further elucidated.

<b>Background:</b> Breast and prostate cancer are frequently diagnosed neoplasias in women and men around the world. The signaling of the androgen receptor (AR) influences the development of both tumors. Since therapies focused to block the receptor's activity have not been fully effective, and have shown side effects, therapies based on natural compounds are promissory complementary alternatives in its treatment. <b>Objective:</b> The aim of this study was to determine the effect of anthocyanins from blue corn in cancer cell lines. <b>Methods:</b> We analyzed the antiproliferative effect of anthocyanins from raw and alkali-processed (tortillas) Mixteco blue corn in breast and prostate cancer cell lines MDA-MB-453 (subtype: triple negative) and LNCaP using methyltiazlyl-tetrazolium (MTT) and flow cytometry (FCM). The combination of anthocyanins and 2-amino-N-quinolin-8-yl-benzenesulfonamide (QBS) or nocodazole also were evaluated. The anthocyanins were isolated trough column chromatography (XAD-7). <b>Results:</b> Our results demonstrated that anthocyanin specially the ones obtained from tortillas, decreased cell viability and arrested cell cycle in G1 phase inducing apoptosis. Cytometry analysis shows an increased effect on apoptosis of MDA-MB-453 and LNCaP cells when tortilla anthocyanins and QBS were combined. <b>Conclusions:</b> This is the first report that suggest that anthocyanins from blue corn have an effect in cell cycle and viability so they could serve as adjuvants for breast and prostate cancer therapies and may prompt to deepen investigations to decipher its molecular properties. Abbreviations AR Androgen Receptor CIDIIR Interdisciplinary Center for Research on Integral Regional Development <em>DHT</em> <em>5α</em>-Dihydrotestosterone ER Estrogen Receptor PR Progesterone Receptor QBS Amino-N-quinolin-8-yl-benzenesulfonamide.

Prostate cancer (PCa) is the second most common type of male malignancy worldwide. The transcription factor zinc finger E‑box binding homeobox 1 (ZEB1) is associated with epithelial‑mesenchymal transition and is also involved in regulation of androgen receptor (AR) expression, the main ligands of which are testosterone and dihydrotestosterone (<em>DHT</em>). These androgens are synthesized through the steroidogenic pathway within the prostate, and their synthesis is altered in PCa. The present study aimed to determine the ZEB1‑induced alterations in androgen synthesis and AR expression in the DU145 PCa cell line. Reverse transcription‑quantitative polymerase chain reaction, western blotting and immunocytochemistry were used to determine the mRNA and protein expression levels, and cellular localization of steroidogenic pathway enzymes in the DU145 cell line in response to ZEB1 silencing. Furthermore, the concentrations of testosterone and <em>DHT</em> were detected in cell culture medium using ELISA. ZEB1‑silenced cells exhibited an increase in testosterone and <em>DHT</em> production, an increase in AR expression and an alteration in the steroidogenic pathway. In particular, steroidogenic acute regulatory protein and <em>5α</em>‑reductase 2 expression levels were decreased, whereas cytochrome P450 family 17 subfamily A member 1, <em>5α</em>‑reductase 1, aldo‑keto reductase family 1 member D1 and aldo‑keto reductase family 1 member C2 expression levels were increased. In conclusion, the present study provided novel information regarding the regulation of intratumoral androgen production in PCa, which is relevant for the progression of the disease to a castration‑resistant form.

Hormones locally synthesized in the anterior pituitary gland are involved in regulation of pituitary cell renewal. In the pituitary, testosterone (T) may exert its actions per se or by conversion to dihydrotestosterone (<em>DHT</em>) or 17β-estradiol (E2) by <em>5α</em>-reductase and aromatase activity, which are expressed in this gland. Previous reports from our laboratory showed that estrogens modulate apoptosis of lactotropes and somatotropes from female rats. Now, we examined the in vitro and in vivo effects of gonadal steroids on apoptosis of anterior pituitary cells from adult male rats. T in vitro did not modify apoptosis in anterior pituitary cells from gonadectomized (GNX) male rats. <em>DHT</em>, a non-aromatizable androgen, exerted direct antiapoptotic action on total anterior pituitary cells and folliculo-stellate cells, but not on lactotropes, somatotropes, or gonadotropes. On the contrary, E2 exerted a rapid apoptotic effect on total cells as well as on lactotropes and somatotropes. Incubation of anterior pituitary cells with T in presence of Finasteride, an inhibitor of <em>5α</em>-reductase, increased the percentage of TUNEL-positive cells. In vivo administration of <em>DHT</em> to GNX rats reduced apoptosis in the anterior pituitary whereas E2 exerted proapoptotic action and reduced cells in G2/M-phase of the cell cycle. In summary, our results indicate that <em>DHT</em> and E2 have opposite effects on apoptosis in the anterior pituitary gland suggesting that local metabolization of T to these steroids could be involved in pituitary cell turnover in males. Changes in expression and/or activity of <em>5α</em>-reductase and aromatase may play a role in the development of anterior pituitary tumors.

Sex hormone deprivation commonly occurs following menopause in women or after androgen-depletion during prostate cancer therapy in men, resulting in rapid bone turnover and loss of bone mass. There is a need to identify novel therapies to improve bone mass in these conditions. Previously, we identified age- and sex-dependent effects on bone mass in mice with deletion of the gene encoding the β-galactoside binding lectin, galectin-3 (Lgals3-KO). Due to the influence of sex on the phenotype, we tested the role of sex hormones, estrogen (β-estradiol; E₂), and androgen (<em>5α</em>-dihydroxytestosterone; <em>DHT</em>) in Lgals3-KO mice. To address this, we subjected male and female wild-type and Lgals3-KO mice to gonadectomy ± E₂ or <em>DHT</em> rescue and compared differential responses in bone mass and bone formation. Following gonadectomy, male and female Lgals3-KO mice had greater cortical bone expansion (increased total area; T.Ar) and reduced loss of bone area (B.Ar). While T.Ar and B.Ar were increased in response to <em>DHT</em> in wild-type mice, <em>DHT</em> did not alter these parameters in Lgals3-KO mice. E₂ rescue more strongly increased B.Ar in Lgals3-KO compared to wild-type female mice due to a failure of E₂ to repress the increase in T.Ar following gonadectomy. Lgals3-KO mice had more osteoblasts relative to bone surface when compared to wild-type animals in sham, gonadectomy, and E₂ rescue groups. <em>DHT</em> suppressed this increase. This study revealed a mechanism for the sex-dependency of the Lgals3-KO aging bone phenotype and supports targeting galectin-3 to protect against bone loss associated with decreased sex hormone production.

Background: Skeletal muscle mass and function are partly maintained by the supply of amino acids, altered amino acid transport is an important cause of frailty that can lead to decreased independence with increasing age and slow trauma recovery. The system-A sodium coupled neutral amino acid transporter (SNAT)-2 coded by gene family SLC38A2 generates a 506 amino acid 56 kDa protein that is an important transporter of amino acids in skeletal muscle. Ageing is associated with a decrease in expression of SNAT2 transporters.

Methods: In this study, we used the C2C12 cell line, using myoblast cells and cells differentiated into myotubes. We investigated if the expression of SNAT2 DNA would enhance intracellular amino acid levels and increase their availability for protein synthesis.

<strong class="sub-title"> Results: </strong> In control myoblasts and myotubes, we found significantly decreased expression of SNAT2 (6.5× decrease, n = 4 per group, P < 0.05) in myotubes than found in myoblasts. After transfection with a SNAT2-eGFP cDNA plasmid, C2C12 myoblasts significantly increased perinuclear punctate SNAT2-eGFP expression that persisted and was more cytoplasmic after differentiation into myotubes. Interestingly, transfected cells were significantly more responsive to the hormone <em>5α</em>-dihydrotestosterone (<em>DHT</em>, 4.5 nM, by 1.6×, n = 3 per group, P < 0.04). Starvation significantly enhanced the amino acid C¹⁴ -MeAIB transport (1.7×, n = 3 per group, P < 0.05) indicating increased function of SNAT2. Inhibiting SNAT2 with high concentrations of MeAIB (3.3 or 5 mM) significantly reduced C¹⁴ -Isoleucine transport by L-type amino acid transporter (LAT2, 52.8% and 77%, respectively, n = 3 per group, P < 0.05). However, there was no increase in the LAT2 transport of C¹⁴ -isoleucine detectable in SNAT2-eGFP transfected cells after <em>DHT</em> (4.5 nM) exposure. This indicated that small amino acid availability was not rate limiting to LAT2 function in myoblasts.

Conclusions: Overall, these data show that transfection of SNAT2-eGFP expression enhanced its function following starvation and treatment with physiological levels of DHT. Enhanced SNAT2 expression in muscle cells offers a viable epigenetic target in pathological conditions associated with altered amino acid transport.

Keywords: Ageing; Amino acid transport; LAT2; Muscle; SNAT2.

Steroid <em>5α</em>-reductase deficiency (5ARD) is a rare autosomal recessive disorder caused by mutation in the <em>5α</em>-reductase type 2 gene (<i>SRD5A2</i>). 5ARD results in the impaired conversion of testosterone (T) to dihydrotestosterone (<em>DHT</em>) and is characterized by undervirilization in 46XY individuals. We report a case series of three siblings presenting with ambiguous genitalia and different phenotypes. They did not meet the widely accepted biochemical criteria for 5ARD. In view of strong clinical suspicion, genetic analysis was performed which revealed pathogenic mutation in <i>SRD5A2</i>. This report highlights the importance of definitive diagnosis with molecular methods as the treatment and prognosis differs greatly among the close differential diagnoses. Reliance on the biochemical criteria alone may lead to misdiagnosis.