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Zhongguo Zhong Yao Za Zhi = Zhongguo... Mar 2021Protein kinase C(PKC) is a kind of kinase which is widely involved in cell proliferation and development. PKC(Wp-PKC) in Whitmania pigra body belongs to classic PKC. In...
Protein kinase C(PKC) is a kind of kinase which is widely involved in cell proliferation and development. PKC(Wp-PKC) in Whitmania pigra body belongs to classic PKC. In order to investigate the effect of Wp-PKC on the development of Wh. pigra germ cells, 17β-estradiol(17β-E2)(100 ng·mL~(-1)) and methyltestosterone(MT)(150 μg·L~(-1)), 150 μg·L~(-1)(MT)+0.5 mg·L~(-1) PKC, 0.5 mg·L~(-1) PKC inhibitor were added to Wh. pigra culture water, and no addition group(control group) was added, and the effects on the development of Wh. pigra germ cells and the expression of Wp-PKC were observed. The results showed that: Wp-PKC in male gonads was always higher than that in female gonads; MT promoted the development of male gonads in Wh. pigra, while the expression of Wp-PKC was significantly higher than that in the control; 17β-E2 promoted the development of female gonads in Wh. pigra and Wp-PKC expression significantly lower than that of the control; while the development of the female and male gonads in the PKC inhibitor group was inhibited, the expression of Wp-PKC was significantly lower than that of the control. In summary, Wp-PKC may promote the development of Wh. pigra, especially the development of male gonads.
Topics: Animals; Estradiol; Female; Gonads; Leeches; Male; Methyltestosterone; Ovary
PubMed: 33787134
DOI: 10.19540/j.cnki.cjcmm.20201224.102 -
Drug Testing and Analysis Jun 2021The knowledge of the biotransformation of compounds prohibited by the World Anti Doping Agency is of high concern as doping analyses are mostly based on the detection of... (Comparative Study)
Comparative Study
The knowledge of the biotransformation of compounds prohibited by the World Anti Doping Agency is of high concern as doping analyses are mostly based on the detection of metabolites instead of the parent compounds abused by athletes. While the self-administration of doping-relevant compounds is from an ethical point of view a rather problematic method to investigate metabolism, the usage of cell culture systems allows for studies on biotransformation in vitro. Five cell culture models with different tissue origin (liver, ovary, skin, kidney, and testis) were comparatively incubated with testosterone and epitestosterone as well as with the synthetic testosterone derivatives 17α-methyltestosterone and 4-chlorotestosterone to investigate the impact of synthetic modifications on phase I metabolic pathways. Cell culture supernatants were analyzed by high-performance liquid chromatography-tandem mass spectrometry. All cell lines possessed the default steroid phase I biotransformation reactions. The highest conversion rate was observed in ovarian (BG-1) and liver cells (HepG2). For BG-1 and skin cells (HaCaT), the 5α-reductase products 5α-dihydrotestosterone (for both) and 5α-androstane-3α/β,17β-diol (for BG-1 solely) were found to be prevailing after testosterone incubation. In kidney (COS-1) and HepG2 cells, the 17β-hydroxysteroid dehydrogenase activity was predominant as supported by the observation that the 17α-OH (epitestosterone) and the methyl group (17α-methyltestosterone) impeded the conversion rate in these cell lines. In conclusion, future work should extend the characterization of the BG-1 and HepG2 cells on phase II metabolic pathways to examine whether they are suitable models for the generation of metabolite reference collections comparable to those obtained by human excretion studies.
Topics: Animals; COS Cells; Cell Culture Techniques; Cell Line; Chlorocebus aethiops; Chromatography, High Pressure Liquid; Doping in Sports; HaCaT Cells; Hep G2 Cells; Humans; Tandem Mass Spectrometry; Testosterone
PubMed: 33527655
DOI: 10.1002/dta.3009 -
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 -
Journal of Fish Biology Jul 2021The cyp11 includes cyp11a and cyp11b in most mammals and teleosts, encoded cholesterol side chain lyase and 11β-hydroxylase, respectively. It is essential in steroid...
The cyp11 includes cyp11a and cyp11b in most mammals and teleosts, encoded cholesterol side chain lyase and 11β-hydroxylase, respectively. It is essential in steroid hormone synthesis. However, studies on the regulation of cyp11 are limited, especially in teleosts. In this study, the molecular characterization and function of cyp11a and cyp11b of black rockfish was investigated. Both of them showed high homology with other teleost counterparts by phylogenetic analysis. The expression of cyp11a and cyp11b exhibited a clear sexually dimorphic pattern, with a higher expression level in testis than that of in ovaries. During the different developmental stages (40 dpf, 80 dpf, 190 dpf, 360 dpf, 720 dpf), the expression of cyp11a was earlier than cyp11b. In situ hybridization results showed that cyp11a and cyp11b were mainly expressed in oogonia and oocytes of the ovary. They were located in spermatogonia and interstitial compartment in the 1.5-year-old gonads, and spermatocytesgonia and the peritubular myoid cell of the testis in the 2.5-year-old gonads. To explore the distinct roles of cyp11a and cyp11b in gonads, oestrogen and androgens were used to stimulate the primary testicular and ovarian cells. The expressions of cyp11a and cyp11b were tested under different dose of 17α-methyltestosterone (17α-MT) and 17β-estradiol (E2). The results showed cyp11a was significantly increased at 10 mol ml of 17α-MT and 10 mol ml of E2 in ovary and 10 mol ml of 17α-MT and E2 in testis, while cyp11b was significantly decreased after 17α-MT and E2 treatment. These results indicated that cyp11a and cyp11b were likely to have different functions, and also implied they might play an important roles in the differentiation of gonads and the synthesis of steroids in black rockfish.
Topics: Animals; Female; Male; Methyltestosterone; Ovary; Perciformes; Phylogeny; Testis
PubMed: 33252824
DOI: 10.1111/jfb.14631 -
Fish Physiology and Biochemistry Feb 2021Gonadotropin-inhibitory hormone (GnIH) plays a critical role in regulating gonadotropin-releasing hormone (GnRH), gonadotropin hormone (GtH), and steroidogenesis. The...
Gonadotropin-inhibitory hormone (GnIH) plays a critical role in regulating gonadotropin-releasing hormone (GnRH), gonadotropin hormone (GtH), and steroidogenesis. The Lpxrfa (the piscine ortholog of GnIH) system has been found to regulate fish reproduction. To gain insight into the role of Lpxrfa in the regulation of spotted scat (Scatophagus argus) reproduction, spotted scat Lpxrfa (ssLpxrfa), and its receptor (ssLpxrfa-r) were cloned and analyzed. Tissue distribution and expression patterns at the hypothalamo-pituitary-gonadal axis (HPG axis) of sslpxrfa and sslpxrfa-r mRNA were also investigated during gonadal development of spotted scat. The open reading frame (ORF) of the sslpxrfa was 606 bp encoding 201 amino acids and includes a putative signal peptide and two mature ssLpxrfa peptides with LPXRFamide motif at their C-terminus. The sslpxrfa-r ORF was 1449 bp encoding 482 amino acids and contracted a seven-hydrophobic transmembrane (TM) domain structure. The tissue distribution showe d that the sslpxrfa was highly expressed in hypothalami, gill, and the gonads. In addition, sslpxrfa-r was highly expressed in hypothalami, pituitaries, and the gonads. Quantitative real-time polymerase chain reaction (qPCR) revealed that sslpxrfa had the highest expression in the hypothalami and pituitaries, and the lowest expression in the gonads in stage V. During gonadal development, the expression of sslpxrfa-r was gradually increased in the hypothalami but reduced in the gonads. However, no obvious trend was observed in the pituitaries. The expression of sslpxrfa and sslpxrfa-r decreased significantly after injection with 17β-estradiol (E). However, the expression of both sslpxrfa and sslpxrfa-r was not changed after injection with 17α-methyltestosterone(17α-MT) in the hypothalami. In addition, no changes were observed in the expression of fshβ and lhβ in the pituitaries after injecting ssLpxrfa-1. However, ssLpxrfa-2 could downregulate the expression of sbgnrh and fshβ in the hypothalami and pituitaries, respectively. Taken together, these findings suggested that ssLpxrfa may participate in E feedback in reproduction and regulate the reproductive axis of spotted scat.
Topics: Amino Acid Sequence; Animals; Estradiol; Female; Fish Proteins; Fishes; Gene Expression Regulation, Developmental; Gonads; Hypothalamo-Hypophyseal System; Hypothalamus; Male; Methyltestosterone; Neuropeptides; Phylogeny; Pituitary Gland; Receptors, Neuropeptide; Reproduction
PubMed: 33215297
DOI: 10.1007/s10695-020-00898-2 -
Steroids Jan 2021There is some evidence that marketable supplements contain hormones not declared on the product label. The presence of these androgenic anabolic steroids (AAS) in sports...
There is some evidence that marketable supplements contain hormones not declared on the product label. The presence of these androgenic anabolic steroids (AAS) in sports supplements can be considered an adulteration and affect the health of consumers, who are predominantly athletes. This study aimed to measure anabolic hormones (methyltestosterone and 4-androstenedione) in sport supplements. Ultra Performance Liquid chromatography coupled mass spectrometry (UPLC-MS/MS) with electrospray ionization (ESI) in positive mode was employed under the Multiple Reaction Monitoring (MRM) ion program. To overcome matrix effects and quantify the selected analyte, the calibration curve was made using Matrix Match method. The LOQ and LOD were 1 ng/g and 0.3 ng/g for both analytes. The recovery of 4-androstenedione and methyltestosterone was in the range of 86.87-107.35 and 77.31-113.98, respectively. In terms of reproducibility, CV % for 4-androstenedione and methyltestosterone ranged from 6.56 to 16.87% and 1.45-15.12%, respectively. 4-androstenedione was found in 11 samples including 9 whey as 1.578 ± 0.154 ng/g and 2 whey albumin samples with an amount of 1.134 ng/g and 1.474 ng/g. Consequently, continuous controlling of sport supplements comprising intentionally or unintentionally added androgens could be important for health and discuss in the context of compliance with anti-doping.
Topics: Androstenedione; Doping in Sports; Methyltestosterone; Reproducibility of Results
PubMed: 33161054
DOI: 10.1016/j.steroids.2020.108758 -
Molecular and Cellular Endocrinology Jan 2021In teleost fish, sex can be determined by genetic factors, environmental factors, or both. Unlike in gonochoristic fish, in which sex is fixed in adults, sex can change... (Review)
Review
In teleost fish, sex can be determined by genetic factors, environmental factors, or both. Unlike in gonochoristic fish, in which sex is fixed in adults, sex can change in adults of hermaphroditic fish species. Thus, sex is generated during the initial gonadal differentiation stage (primary sex differentiation) and later during sexual fate alternation (secondary sex differentiation) in hermaphroditic fish species. Depending on the species, sex phase alternation can be induced by endogenous cues (such as individual age and body size) or by social cues (such as sex ratio or relative body size within the population). In general, the fluctuation in plasma estradiol-17β (E2) levels is correlated with the sexual fate alternation in hermaphroditic fish. Hormonal treatments can artificially induce sexual phase alternation in sequential hermaphroditic fishes, but in a transient and reversible manner. This is the case for the E2-induced female phase in protandrous black porgy and the methyltestosterone (MT)- or aromatase inhibitor (AI)-induced male phase in protogynous grouper. Recent reviews have focused on the different forms of sex change in fish who undergo sequential sex change, especially in terms of gene expression and the role of hormones. In this review, we use the protandrous black porgy, a nonsocial cue-influenced hermaphroditic species, with digonic gonads (ovarian and testis separated by a connective tissue), as a model to describe our findings and discuss the molecular and cellular regulation of sexual fate determination in hermaphroditic fish.
Topics: Animals; Aquaculture; Disorders of Sex Development; Female; Male; Models, Animal; Models, Biological; Perciformes; Sex Differentiation
PubMed: 33127483
DOI: 10.1016/j.mce.2020.111069 -
Steroids Dec 2020This paper deals with quantification of the following steroids in mixture: hydrocortisone (1), deoxycorticosterone (2), progesterone (3) and methyltestosterone (4) by...
This paper deals with quantification of the following steroids in mixture: hydrocortisone (1), deoxycorticosterone (2), progesterone (3) and methyltestosterone (4) by means of mass spectrometry and implementing our innovative stochatic dynamic functional relationship between the analyte concentration in solution and the experimental variable intensity. The mass spectrometric data are correlated independently using chromatography. Chemometric analysis is carried out.
Topics: Complex Mixtures; Mass Spectrometry; Steroids; Stochastic Processes
PubMed: 33069721
DOI: 10.1016/j.steroids.2020.108750 -
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 -
Brazilian Journal of Biology = Revista... 2021Synthetic androgens (male hormones) administered to fish nursery are being used in aquaculture to avoid sexual differentiation and unwanted spawning at the eggs or the...
Synthetic androgens (male hormones) administered to fish nursery are being used in aquaculture to avoid sexual differentiation and unwanted spawning at the eggs or the first feeding fry stage of fish. Present trial was conducted with the aim to produce male common carp (Cyprinus carpio) by egg immersion technique. Through this little insight, the effect of different hormone concentrations (17α-methyltestosterone @ HC:150, 300, 450 and 600 µgl-1) with immersion times (IT: 24, 48 and 72 hrs) and their interaction effect (HC x IT) on the hatching percentage of Cyprinus carpio eggs, percent survival and percent of male's production was evaluated specifically. Results showed that egg hatching percentage decreased with increased IT likewise, survival of treated fry was affected by increasing the IT (P<0.001). The main interaction effect of HC x IT showed that the highest percent of male individuals (95%) was obtained at 450-600 µgl-1 HC for 72 hrs IT, followed by 88-92.50% at 150-300 µgl-1 HC for 72-hrsof IT, 87.50% at 48-hrs of IT for rest of the hormone treatments, and lowest 47.50% was recorded in control (P<0.05). Increased percent male of Cyprinus carpio was obtained with increasing HC across all ITs. It was observed that the immersion treatment at 600µgl-1 for 72 hours was more effective to change the sex ratio of pre hatch Cyprinus carpio. A comparative outlook made from this experimental trial that sex induction of Cyprinus carpio by eggs immersion using synthetic male steroid hormone is an alternative safe technique of fish sex reversal in contrast to oral administration of hormone in fish feed.
Topics: Androgens; Animals; Aquaculture; Carps; Immersion; Male
PubMed: 32901653
DOI: 10.1590/1519-6984.224681