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Journal of Chromatography. A Jul 2021The research was done with partial filling micellar electrokinetic chromatography, microemulsion electrokinetic chromatography, and ultra-high performance liquid...
The research was done with partial filling micellar electrokinetic chromatography, microemulsion electrokinetic chromatography, and ultra-high performance liquid chromatography. The study focuses on determination of male and female steroids from cold and hot tap water of households in Helsinki City. The district´s raw water is made run from Päijänne Lake through a water tunnel to the purification plants in Helsinki area. The effluents delivered from the plants to households as tap water were sampled and used for the study. They were concentrated with solid phase extraction to exceed the detection limits of the three methods. With partial filling method the limits were 0.50, 0.48, 0.33, and 0.50 mg/L for androsterone, testosterone, progesterone, and testosterone-glucuronide, respectively. In microemulsion method the limit values were 1.33, 1.11, and 0.40 mg/L for androsterone, testosterone, and progesterone, respectively, and 0.83, 0.45, and 0.50 mg/L for hydrocortisone, 17-α-hydroxyprogesterone, and 17-α-methyltestosterone, respectively. In the tap water samples, progesterone concentrations represented the highest values being 0.22 and 1.18 ng/L in cold and hot water, respectively. They also contained testosterone (in all samples), its glucuronide metabolite (in 25% of the samples), and androstenedione (in 75% of the samples). The ultra-high liquid chromatographic method with mass spectrometric detection was used for identification of the steroids at µg/L level.
Topics: Chromatography, Liquid; Electrophoresis, Capillary; Gonadal Steroid Hormones; Lakes; Limit of Detection; Mass Spectrometry; Solid Phase Extraction; Steroids; Water; Water Pollutants, Chemical
PubMed: 34038782
DOI: 10.1016/j.chroma.2021.462233 -
The Journal of Steroid Biochemistry and... Dec 2019Estrogen receptor (ER) sequences vary between species and this suggests that there are differences in the ligand-specificity, leading to species-specific effects. This...
Estrogen receptor (ER) sequences vary between species and this suggests that there are differences in the ligand-specificity, leading to species-specific effects. This would indicate that it is not possible to generalize effects across species. In this study, we investigated the differences in activation potencies and binding affinities of ER´s alpha (α) and beta (β) in human, zebrafish and sea bream to elucidate species differences in response to estradiol, estrone, estriol and methyltestosterone. In vitro analysis showed that estradiol had the highest activity for all the ER´s except for human ERβ and seabream ERβ2. Alignment of the ligand binding domain and ligand binding pocket (LBP) residues of the three species showed that different residues were involved in the LBPs which led to differences in pocket volume, affected binding affinity and orientation of the ligands. By combining in silico and in vitro results, it was possible to identify the ligand specificities of ER´s. The results demonstrated that the human ER´s show lower resolution in ligand-dependent activation, suggesting higher promiscuity, than the zebrafish and seabream ER´s. These results show species-specificity of ER´s and suggest that species-specific differences must be taken into consideration when studying different exposure scenarios.
Topics: Androgens; Animals; Cell Line; Estradiol; Estriol; Estrogens; Estrone; Fish Proteins; Humans; Ligands; Methyltestosterone; Molecular Docking Simulation; Receptors, Estrogen; Sea Bream; Species Specificity; Zebrafish
PubMed: 31437548
DOI: 10.1016/j.jsbmb.2019.105450 -
Biology of Reproduction May 2020Genital tubercle has bisexual potential before sex differentiation. Females exposed to androgen during sex differentiation show masculinized external genitalia, but the...
Genital tubercle has bisexual potential before sex differentiation. Females exposed to androgen during sex differentiation show masculinized external genitalia, but the effects of different androgens on tubular urethral and penile formation in females are mostly unknown. In this study, we compared the masculinization effects of commonly used androgens methyltestosterone, dihydrotestosterone, and testosterone on the induction of penile formation in females. Our results suggested that prenatal treatment with low doses of methyltestosterone, but not same doses of dihydrotestosterone or testosterone, could induce penile formation in female mice. The minimum dose of dihydrotestosterone and testosterone for inducing tubular urethral formation in female mice was, respectively, 50 and 20 times higher than that of methyltestosterone. In vivo methyltestosterone treatment induced more nuclear translocation of androgen receptors in genital tubercles of female mice, affected Wnt signaling gene expressions, and then led to similar patterns of cell proliferation and death in developing genital tubercles to those of control males. We further revealed that low-dose methyltestosterone, but not same dose of dihydrotestosterone or testosterone, treatment induced penile formation in female guinea pigs. Exposure of female mouse genital tubercle organ culture to methyltestosterone, dihydrotestosterone, or testosterone could induce nuclear translocation of androgen receptors, suggesting that the differential effect of the three androgens in vivo might be due to the hormonal profile in mother or fetus, rather than the local genital tissue. To understand the differential role of these androgens in masculinization process involved is fundamental to androgen replacement therapy for diseases related to external genital masculinization.
Topics: Active Transport, Cell Nucleus; Androgen Antagonists; Animals; Animals, Newborn; Cell Death; Cell Proliferation; Dihydrotestosterone; Drug Therapy, Combination; Female; Flutamide; Genitalia, Female; Guinea Pigs; Male; Methyltestosterone; Mice; Penis; Pregnancy; Receptors, Androgen; Sex Determination Analysis; Testosterone
PubMed: 32219310
DOI: 10.1093/biolre/ioaa035 -
The Journal of Steroid Biochemistry and... Sep 2024Methyltestosterone (MT) is one of the most frequently misused anabolic androgenic steroids detected in doping control analysis. The metabolism of MT in humans leads to...
Methyltestosterone (MT) is one of the most frequently misused anabolic androgenic steroids detected in doping control analysis. The metabolism of MT in humans leads to several phase І metabolites and their corresponding phase Ⅱ conjugates. Previous studies have postulated the 3α-sulfoconjugate of 17α-methyl-5β-androstane-3α,17β-diol (S2) as principal sulfate metabolite of MT, with a detection window exceeding 10 days. However, a final direct and unambiguous confirmation of the structure of this metabolite is missing until now. In this study, we established an approach to detect and identify S2, using intact analysis by liquid chromatography hyphenated with tandem mass spectrometry (LC-MS/MS) without complex sample pretreatment. An in vitro study yielded the LC-MS/MS reference retention times of all 3-sulfated 17-methylandrostane-3,17-diol diastereomers, allowing for accurate structure assignment of potentially detected metabolites. In an in vivo excretion study with a single healthy male volunteer, the presence of the metabolite S2 was confirmed after a single oral dose of 10 mg MT. The reference standard was chemically synthesized, characterized by accurate mass mass spectrometry (MS) and nuclear magnetic resonance (NMR), and quantified by quantitative NMR (qNMR). Thus, this study finally provides accurate structure information on the S2 metabolite and a direct analytical method for detection of MT misuse. The availability of the reference material is expected to facilitate further evaluation and subsequent analytical method validation in anti-doping research.
Topics: Male; Humans; Methyltestosterone; Tandem Mass Spectrometry; Chromatography, Liquid; Substance Abuse Detection; Doping in Sports; Anabolic Agents; Adult; Liquid Chromatography-Mass Spectrometry
PubMed: 38710312
DOI: 10.1016/j.jsbmb.2024.106527 -
Molecules (Basel, Switzerland) Oct 2022Molecularly imprinted polymers@ethylenediamine-modified carbon dots grafted on cotton fabrics (MIPs@EDA-CDs/CF) and smartphone-based fluorescence image analysis were...
Molecularly imprinted polymers@ethylenediamine-modified carbon dots grafted on cotton fabrics (MIPs@EDA-CDs/CF) and smartphone-based fluorescence image analysis were proposed and used for the first time for the detection of 17 α-methyltestosterone (MT). The EDA-CDs were synthesized and grafted on cotton fabric before coating with the MIPs. The MIPs were synthesized using the MT as a template molecule, methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a cross-linker, and azobisisobutyronitrile (AIBN) as an initiator. The MIPs@EDA-CDs/CF were characterized using FTIR, SEM-EDS, and RGB fluorescence imaging. The fluorescence images were also taken using a smartphone and the ImageJ program was used for RGB measurement. The Δ red intensity was linearly proportional to MT concentration in the range of 100 to 1000 μg/L (R = 0.999) with a detection limit of 44.4 μg/L and quantification limit of 134 μg/L. The MIPs@EDA-CDs/CF could be stored at 4 °C for a few weeks and could be reused twice. The proposed method could apply for the specific determination of MT in water and sediment samples along with satisfactory recoveries of 96-104% and an acceptable relative standard deviation of 1-6% at the ppb level.
Topics: Molecular Imprinting; Carbon; Molecularly Imprinted Polymers; Methyltestosterone; Polymers
PubMed: 36364082
DOI: 10.3390/molecules27217257 -
Animals : An Open Access Journal From... Jan 2024The average daily weight gain (ADG) is considered a crucial indicator for assessing growth rates in the swine industry. Therefore, investigating the gastrointestinal...
The average daily weight gain (ADG) is considered a crucial indicator for assessing growth rates in the swine industry. Therefore, investigating the gastrointestinal microbiota and serum metabolites influencing the ADG in pigs is pivotal for swine breed selection. This study involved the inclusion of 350 purebred Yorkshire pigs (age: 90 ± 2 days; body weight: 41.20 ± 4.60 kg). Concurrently, serum and fecal samples were collected during initial measurements of blood and serum indices. The pigs were categorized based on their ADG, with 27 male pigs divided into high-ADG (HADG) and low-ADG (LADG) groups based on their phenotype values. There were 12 pigs in LADG and 15 pigs in HADG. Feces and serum samples were collected on the 90th day. Microbiome and non-targeted metabolomics analyses were conducted using 16S rRNA sequencing and liquid chromatography-mass spectrometry (LC-MS). Pearson correlation, with Benjamini-Hochberg (BH) adjustment, was employed to assess the associations between these variables. The abundance of and in LADG was significantly higher than in HADG, while , , , , and in LADG were significantly lower than in HADG. The concentration of glutamine, etiocholanolone glucuronide, and retinoyl beta-glucuronide in LADG was significantly higher than in HADG, while arachidonic acid, allocholic acid, oleic acid, phenylalanine, and methyltestosterone in LADG were significantly lower than in HADG. The - networks (, , methyltestosterone, phenylalanine, oleic acid, arachidonic acid, glutamine, 3-ketosphingosine, L-octanoylcarnitine, camylofin, 4-guanidinobutyrate 3-methylcyclopentadecanone) were identified as the most influential at regulating swine weight gain. These findings suggest that the gastrointestinal tract regulates the daily weight gain of pigs through the network of and . However, this study was limited to fecal and serum samples from growing and fattening boars. A comprehensive consideration of factors affecting the daily weight gain in pig production, including gender, parity, season, and breed, is warranted.
PubMed: 38254447
DOI: 10.3390/ani14020278 -
Cancer Research Communications Jul 2022Inhibiting the androgen receptor (AR), a ligand-activated transcription factor, with androgen deprivation therapy is a standard-of-care treatment for metastatic prostate...
UNLABELLED
Inhibiting the androgen receptor (AR), a ligand-activated transcription factor, with androgen deprivation therapy is a standard-of-care treatment for metastatic prostate cancer. Paradoxically, activation of AR can also inhibit the growth of prostate cancer in some patients and experimental systems, but the mechanisms underlying this phenomenon are poorly understood. This study exploited a potent synthetic androgen, methyltestosterone (MeT), to investigate AR agonist-induced growth inhibition. MeT strongly inhibited growth of prostate cancer cells expressing AR, but not AR-negative models. Genes and pathways regulated by MeT were highly analogous to those regulated by DHT, although MeT induced a quantitatively greater androgenic response in prostate cancer cells. MeT potently downregulated DNA methyltransferases, leading to global DNA hypomethylation. These epigenomic changes were associated with dysregulation of transposable element expression, including upregulation of endogenous retrovirus (ERV) transcripts after sustained MeT treatment. Increased ERV expression led to accumulation of double-stranded RNA and a "viral mimicry" response characterized by activation of IFN signaling, upregulation of MHC class I molecules, and enhanced recognition of murine prostate cancer cells by CD8 T cells. Positive associations between AR activity and ERVs/antiviral pathways were evident in patient transcriptomic data, supporting the clinical relevance of our findings. Collectively, our study reveals that the potent androgen MeT can increase the immunogenicity of prostate cancer cells via a viral mimicry response, a finding that has potential implications for the development of strategies to sensitize this cancer type to immunotherapies.
SIGNIFICANCE
Our study demonstrates that potent androgen stimulation of prostate cancer cells can elicit a viral mimicry response, resulting in enhanced IFN signaling. This finding may have implications for the development of strategies to sensitize prostate cancer to immunotherapies.
Topics: Male; Humans; Animals; Mice; Receptors, Androgen; Androgens; Prostatic Neoplasms; Androgen Antagonists; CD8-Positive T-Lymphocytes; DNA
PubMed: 36923279
DOI: 10.1158/2767-9764.CRC-21-0139 -
Physiology & Behavior Jan 2021Fishes can change their physiological responses when threatened by the presence of predators. Such physiological plasticity, however, usually implies costs that may...
Fishes can change their physiological responses when threatened by the presence of predators. Such physiological plasticity, however, usually implies costs that may impede organismal development and reproduction and reduce the ability to cope with other biotic and abiotic stresses. Here, we evaluated the growth and stress biomarker responses in sexually reversed Nile tilapia, Oreochromis niloticus, fingerlings indirectly threatened by the presence of the aquatic insect predator Belostoma anurum (Hemiptera: Belostomatidae). We also evaluated whether the presence of B. anurum would affect growth in fingerlings that received food containing the masculinizing hormone 17 α-methyltestosterone. The antioxidant responses were evaluated by measuring the activity of enzymes (e.g., superoxide dismutase, catalase, and glutathione-S-transferase). Oxidative stress biomarkers (e.g., malondialdehyde and nitric oxide) and blood glucose and lactate responses were also evaluated. Our results revealed that predator exposure did not affect growth in O. niloticus fingerlings reared in the presence or absence of the masculinizing hormone. However, sexually reversed tilapia fingerlings significantly increased not only the glucose and lactate blood levels, but also exhibited increased activities of superoxide dismutase and glutathione-S-transferases enzymes when threatened by the presence of B. anurum nymphs. Collectively, our findings indicate that despite not exhibiting reduced growth performance, sexually reversed tilapia fingerlings were physiologically stressed by the presence of the predator, which may reduce their ability to face environmental and abiotic stresses.
Topics: Animals; Antioxidants; Biomarkers; Cichlids; Insecta; Liver; Oxidative Stress
PubMed: 33039381
DOI: 10.1016/j.physbeh.2020.113202 -
Animals : An Open Access Journal From... Apr 2023The steroid 17α-methyltestosterone (MT) inhibits ovarian function and is often used to induce sex reversal artificially in vertebrates. In the present study, different...
The steroid 17α-methyltestosterone (MT) inhibits ovarian function and is often used to induce sex reversal artificially in vertebrates. In the present study, different concentrations of MT were added as dietary supplementation, and the effects on sex ratio, growth, and gonadal development were examined. After 40 days, the sex ratio (male:female) in each group increased at different degrees with 50 (1.36:1), 100 (1.57:1), and 200 (2.61:1) mg/kg MT, and neo-males with testis-ovary coexistence were observed in the 200 mg/kg MT group. Furthermore, 50 and 100 mg/kg MT could induce female reversion in neo-males. Histologically, the development of the testes in experimental groups was slower, but the ovaries of the experimental and control groups had similar developmental rates. The expression levels of , , and in males at 200 mg/kg MT were 8.65-, 3.75-, and 3.45-fold greater than those of the control group. In crustaceans, sex reversal through vertebrate sex hormones can be observed. Neo-males (sex-reversed female prawns) were maintained by exogenous androgen, and over-reliance led to slow testis growth, small body size, and low growth rate, but sperm was still produced. In female prawns, MT inhibited ovary development and promoted growth.
PubMed: 37106932
DOI: 10.3390/ani13081369 -
International Journal of Molecular... Aug 2019A-type K channels contribute to regulating the propagation and frequency of action potentials in smooth muscle cells (SMCs). The present study (i) identified the...
A-type K channels contribute to regulating the propagation and frequency of action potentials in smooth muscle cells (SMCs). The present study (i) identified the molecular components of A-type K channels in rat vas deferens SMs (VDSMs) and (ii) showed the long-term, genomic effects of testosterone on their expression in VDSMs. Transcripts of the A-type K channel α subunit, Kv4.3L and its regulatory β subunits, KChIP3, NCS1, and DPP6-S were predominantly expressed in rat VDSMs over the other related subtypes (Kv4.2, KChIP1, KChIP2, KChIP4, and DPP10). A-type K current (I) density in VDSM cells (VDSMCs) was decreased by castration without changes in I kinetics, and decreased I density was compensated for by an oral treatment with 17α-methyltestosterone (MET). Correspondingly, in the VDSMs of castrated rats, Kv4.3L and KChIP3 were down-regulated at both the transcript and protein expression levels. Changes in Kv4.3L and KChIP3 expression levels were compensated for by the treatment with MET. These results suggest that testosterone level changes in testosterone disorders and growth processes control the functional expression of A-type K channels in VDSMCs.
Topics: Animals; Blotting, Western; Castration; Down-Regulation; Electrophysiology; Kv Channel-Interacting Proteins; Male; Methyltestosterone; Muscle, Smooth; Rats; Rats, Wistar; Testosterone; Vas Deferens
PubMed: 31438481
DOI: 10.3390/ijms20174073