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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 -
Molecules (Basel, Switzerland) Sep 2017The catalytic activity of enzymes produced by an entomopathogenic filamentous fungus ( KCh J2) towards selected steroid compounds (androstenedione, adrenosterone,...
The catalytic activity of enzymes produced by an entomopathogenic filamentous fungus ( KCh J2) towards selected steroid compounds (androstenedione, adrenosterone, progesterone, 17α-methyltestosterone and dehydroepiandrosterone) was investigated. All tested substrates were efficiently transformed. The structure of the substrate has a crucial impact on regio- and stereoselectivity of hydroxylation since it affects binding to the active site of the enzyme. Androstenedione was hydroxylated in the 7α-position to give a key intermediate in the synthesis of the diuretic-7α-hydroxyandrost-4-ene-3,17-dione with 82% conversion. Adrenosterone and 17α-methyltestosterone were hydroxylated in the 6β-position. Hydroxylated derivatives such as 15β-hydroxy-17α-methyltestosterone and 6β,12β-dihydroxy-17α-methyltestosterone were also observed. In the culture of KCh J2, DHEA was effectively hydroxylated in the C-7 position and then oxidized to give 7-oxo-DHEA, 3β,7α- and 3β,7β-dihydroxy-17a-oxa-d-homo-androst-5-ene-17-one. We obtained 7β-OH-DHEA lactone with 82% yield during 3 days transformation of highly concentrated (5 g/L) DHEA.
Topics: Androstenedione; Androstenes; Animals; Biocatalysis; Biotransformation; Cordyceps; Dehydroepiandrosterone; Fungal Proteins; Hydroxylation; Lactones; Methyltestosterone; Molecular Structure; Progesterone; Spiders; Substrate Specificity
PubMed: 28891949
DOI: 10.3390/molecules22091511 -
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 -
The Journal of Veterinary Medical... Mar 2021Urethral sphincter mechanism incompetence (USMI) is a common cause of urinary incontinence in dogs. Although estrogen is often prescribed for the medical therapy of USMI...
Urethral sphincter mechanism incompetence (USMI) is a common cause of urinary incontinence in dogs. Although estrogen is often prescribed for the medical therapy of USMI for spayed female dogs, they are known to have limited effectiveness and potential adverse effects. In castrated male dogs with USMI, testosterone reagents have been attempted besides estrogen. In this study, the effect of testosterone drugs, mainly methyltestosterone, on spayed female dogs with USMI was retrospectively evaluated. Ten spayed female dogs with USMI were included. Diagnosis of USMI was based on the results of the dogs' medical history, clinical signs, and no abnormalities in physical examinations, urinalysis, ultrasonography, X-ray imaging, and neurological examinations. Methyltestosterone was administered at doses of 0.32-1.27 mg/kg BW p.o. semel in die (sid.) to twice a week. Nine of the ten dogs had good or excellent responses 2 to 4 weeks after the start of treatment. The minimum effective dose was 0.32 mg/kg/day. Although no severe adverse symptoms occurred in any dog, a mild increase in alanine aminotransferase was temporally observed at doses of 1.0 and 1.1 mg/kg/day in the two dogs. After dose reduction or withdrawal, two of eight dogs had recurrence of urinary incontinence. Resumption of testosterone treatment clearly improved the symptoms in the two dogs. These results indicate that testosterone reagents might be an option for treating USMI in spayed female dogs as well.
Topics: Animals; Dog Diseases; Dogs; Female; Male; Retrospective Studies; Testosterone; Urethra; Urinary Incontinence
PubMed: 33441521
DOI: 10.1292/jvms.20-0515 -
International Journal of Molecular... Jan 2024This study investigates the role of lysosomal acid lipase in sex hormone regulation and gonadal development in . The full-length cDNA was cloned, and its expression...
This study investigates the role of lysosomal acid lipase in sex hormone regulation and gonadal development in . The full-length cDNA was cloned, and its expression patterns were analyzed using quantitative real-time PCR (qPCR) in various tissues and developmental stages. Higher expression levels were observed in the hepatopancreas, cerebral ganglion, and testes, indicating the potential involvement of in sex differentiation and gonadal development. In situ hybridization experiments revealed strong signaling in the spermatheca and hepatopancreas, suggesting their potential role in steroid synthesis (such as cholesterol, fatty acids, cholesteryl ester, and triglycerides) and sperm maturation. Increased expression levels of male-specific genes, such as insulin-like androgenic gland hormone , sperm gelatinase , and mab-3-related transcription factor , were observed after (double-stranded ) injection, and significant inhibition of sperm development and maturation was observed histologically. Additionally, the relationship between and sex-related genes (, , and ) and hormones (17β-estradiol and 17α-methyltestosterone) was explored by administering sex hormones to male prawns, indicating that does not directly control the production of sex hormones but rather utilizes the property of hydrolyzing triglycerides and cholesterol to provide energy while influencing the synthesis and secretion of self-sex hormones. These findings provide valuable insights into the function of in and its potential implications for understanding sex differentiation and gonadal development in crustaceans. It provides an important theoretical basis for the realization of a monosex culture of .
Topics: Animals; Male; Palaemonidae; Semen; Gonadal Steroid Hormones; Cholesterol; Triglycerides; Arthropod Proteins
PubMed: 38338678
DOI: 10.3390/ijms25031399