Computer Based Screening of Selected Phytoconstituents from Cyperus Rotundus Linn. Against 5 α Reductase Enzyme

Hirsutism is a condition of unwanted, male-pattern terminal hair growth in women. Electro epilation, laser treatment, intense pulsed light therapy, eflornithine cream, and oral antiandrogen medications are the various allopathic therapeutics have been employed for treatment of Hirsutism. However, a significant number of patients experience discomfort with reported procedures. Cyperus Rotundus Linn. have reported for its anti-androgenic activity effective against Hirsutism disorder. The main objective of present investigation is to screen the selected phytoconstituents of stated plant against 5 α Reductase Enzyme. In this study Cyperene, humulen, β- selinene, campholenic aldehyde, and α-pinene were docked with 5 α Reductase Enzyme using PyRx 0.8.Autodock and binding energies were obtained. The present investigation concludes that the molecular docking analysis of selected phytoconstituents with 5 α reductase enzyme shows good interaction. The binding affinity of Humulen is higher than others whereas campholenic aldehyde showed lowest affinity amongst all other constituents. Further studies need to be performed at laboratory level to support results of computational screening of present investigation.

Androstenedione Is the Preferred Substrate for Cytochrome P450 11β-hydroxylase Leading to the Production of 11β-Hydroxyandrostenedione in the Adrenal Gland

Adrenal cytochrome P450 11β-hydroxylase (CYP11B1) is a mitochondrial enzyme that catalyzes the final step of glucocorticoid synthesis, converting 11-deoxycortisol (S) and deoxycorticosterone (DOC) to cortisol (F) and corticosterone (CORT), respectively. CYP11B1 is predominantly located in the zona fasciculata of the adrenal gland with studies also showing that CYP11B1 catalyzes the conversion of androstenedione (A4) and testosterone (T) to 11β-hydroxyandrostenedione (11OHA4) and 11β-hydroxytestosterone (11OHT), respectively. Adrenal vein sampling in women has shown that 11OHA4 turnover is high, with a marked increase after treatment with ACTH which is known to upregulate CYP11B1 expression. We hypothesized that CYP11B1’s affinity for A4 as a substrate may be favored over glucocorticoids since 11OHA4 is one of the major androgens produced in the adrenalThis study aimed to elucidate the kinetic parameters (Km and Vmax) for A4 and T with respect to CYP11B1. A4 and T (0.2 µM to 5 µM) were assayed in HEK-293 cells transiently transfected with CYP11B1 and the adrenodoxin redox partner. Data was used to generate progress curves and fitted to the Michaelis-Menten equation. A4 had the lowest Km (0.21 μM) with a significantly higher Vmax (315.77 pmol/min/mg protein) in comparison to T, S and DOC. Results suggest that A4 binds more readily to CYP11B1 resulting in the high turnover of substrate. The androgenic activity of CYP11B1 catalyzed steroid products was determined using a luciferase assay conducted in CV1 cells. Both A4 and 11OHA4 showed no androgenic activity, however, the 11βHSD2 product of 11OHA4, 11-ketoandrostenedione (11KA4), elicited a response at 100 nM. 11OHT was an androgen receptor (AR) agonist, however, exhibiting a lower response when compared to T and 11-ketotestosterone over all concentrations tested (1, 10 and 100 nM). As confirmation of CYP11B1 activity in the adrenal, circulatory steroid levels were determined in healthy female subjects (n=88). CORT, F and 11OHA4 were measured at a frequency of 70–100%, compared to 11OHT (40–70%), while 11βHSD2 activity produced 11KA4 (<10%). In conclusion, the catalytic efficiency of CYP11B1 towards A4 is higher compared to T and the classical substrates. The high substrate affinity and turnover provides evidence for 11OHA4 in adrenal vein samples. Concurrently, our analysis suggests that agonism is diminished by the presence of the C11- hydroxyl group, while AR agonistic activity is gained upon its conversion to a keto group. Furthermore, these androgens could perhaps modulate cortisol production in the adrenal due to potential competition between precursor substrates.

Disruption of Estrogenic and Androgenic Bioactivities in Human Fetuses Exposed to Maternal Smoking

Endocrine disruptors (EDs) interfere with hormonal signalling and, given that multiple developmental processes are hormone-driven, the prenatal period is a window of increased sensitivity. Maternal smoking is a real-life model of in utero exposure to a complex mixture of EDs. Cigarette smoke contains of >7,000 pollutants, including polycyclic aromatic hydrocarbons (PAHs), which are AhR ligands and cross-talk with the estrogen receptor (ER) system. Prenatal exposure to cigarette smoke is associated with adverse outcomes, including intrauterine growth restriction and increased risk of metabolic syndrome later in life. We aimed to evaluate ED effects associated with smoke exposure in human fetuses. Fetal tissues (plasma, n=48; placenta, n=30; liver, n=29) from elective terminations of normally progressing pregnancies, ranging from 10 to 20 gestation weeks, were collected (SAFeR and FEGO studies: REC 15/NS/0123, REC 04/S0802/21). PAHs and PAH-like compounds were extracted from placenta and fetal liver. Bioactivity levels in plasma, placenta and liver extracts were determined using ER and androgen receptor (AR) transactivation reporter gene assays. PAH burden was evaluated using the AhR-responsive DRhp-CALUX assay. Smoke exposure was associated with a 1.3-fold increase in plasma estrogenic activity. The developmental trajectory of androgenic activity was altered in plasma of smoke-exposed fetuses, with significant anti-androgenic activity in older fetuses (>16 weeks of gestation). In males, plasma androgenic activity was positively associated with testes weight and anogenital distance. In contrast, placentas from smoking mothers had significantly increased androgenic potential. Furthermore, AhR-like activity was 2.9-fold higher in smoke-exposed placentas compared to controls, and 2.3-fold higher in female compared to male fetal livers. Overall, all bioactivity levels were higher in placentas compared to fetal liver. Prenatal exposure to cigarette smoke is associated with higher placental AhR activation, indicative of increased xenotoxicants burden. We also report that smoke-exposed fetuses showed increased circulating estrogenic activity and disrupted androgenic potential, across 10-20 weeks of gestation, in both fetal plasma and placenta. This demonstrates that EDs present in cigarette smoke are able to interfere with hormonal signalling and alter dynamic endocrine activity profiles, which are critical to ensure appropriate, sex-specific, development. These ED effects are likely to disturb placental function and reprogramme fetal development and thus impacting on life-long health.

Bioanalytical Detection of Steroid Abuse in Sports Based on the Androgenic Activity Measurement

The anabolic androgenic steroids (AAS) are the most frequently consumed performance enhancing drugs (PED) in sports. In the anti-doping field, the detection of AAS is carried out by the analysis of the athlete’s urine using methodologies based on liquid/gas chromatography-mass spectrometry. Unfortunately, the detection of unknown compounds is not possible. BDS’s AR CALUX® bio detection technology was studied as an indirect method to detect the administration of a single dose of testosterone (T). Twelve T and placebo single dose administered men volunteers underwent a triple-blind crossover clinical trial. The UGT2B17 deletion was present among the volunteers and evenly distributed in heterozygous (ins/del), wild-type homozygous (ins/ins), and mutated homozygous (del/del) groups. A significant statistical difference in terms of bioluminescence was observed after the testosterone (T) administration for the three types of polymorphic groups. The ratio of means between the pre- and post-T administration periods, depending on the type of polymorphism, was in group ins/ins 3.31 (CI. 95%: 2.07–5.29), group ins/del 4.15 (CI 95%: 3.05–5.67), and group del/del 2.89 (CI 95%: 2.42–3.46). The results of the study are very promising, as they may offer us the possibility of designing a detection approach that, based on intra-individual monitoring of androgenic values, in the UGT2B17 deletion type.

Hirsutism Caused by Pregnancy Luteoma in a Low-Resource Setting: A Case Report and Literature Review

Background. Pregnancy luteomas are rare, benign, ovarian neoplasms resulting from increased androgenic activity during pregnancy. Often, they occur asymptomatically and are only diagnosed incidentally during imaging or surgery: cesarean section or postpartum tubal ligation. Most common symptoms associated with pregnancy luteoma include acne, deepening of voice, hirsutism, and clitoromegaly. Most pregnancy luteomas regress spontaneously postpartum. Thus, the management of pregnancy luteomas depends on the clinical situation. Case. We report a case of 28-year-old gravida 2, para 1 who presented at 39 + 1 weeks of gestation with prolonged labor and delivered by emergency cesarean. Intraoperatively, a huge left ovarian mass was identified and resected, and tissue was sent for histopathology and a diagnosis of pregnancy luteoma was made after the pathological report. Conclusion. The present report emphasizes that pregnancy luteoma is a benign neoplasm and imprudent surgical intervention should be reserved. Proper imaging techniques, preferably MRI or ultrasonography that visualize the size of the ovary and reproductive hormonal profiles, would suffice for the diagnosis and management of pregnancy luteoma.

Exposure to non-persistent pesticides and puberty timing: a systematic review of the epidemiological evidence

Background: Numerous modern non-persistent pesticides have demonstrated estrogenic/anti-androgenic activity and have been classified as endocrine disrupting chemicals (EDCs). Processes involved in puberty development are vulnerable to EDCs, such as compounds that interfere with the metabolism or activity of sex steroids. Objective: To conduct a systematic review of epidemiological studies on the relationship between early-life exposure to non-persistent pesticides and puberty timing and/or sexual maturation in girls and boys. Methods: A systematic search was carried out using MEDLINE and SCOPUS databases, including original articles published up to November 2020. Results: Thirteen studies were selected after excluding non-original and non-human studies. Exposure to different types of pesticides has been associated with altered puberty timing in girls and/or boys in eight studies. In utero exposure to atrazine has been related to earlier age of menarche in girls; exposure to organophosphate (OP) pesticides has been related to delayed sexual development in boys and girls; childhood pyrethroid exposure has been associated with pubertal delay in girls and pubertal advancement in boys; and prenatal/childhood exposure to multiple pesticides has been linked to earlier puberty onset in girls and pubertal delay in boys. Conclusions: Most of the reviewed studies describe a relationship between pesticide exposure and changes in the age of puberty onset or sex hormone levels, although the quality of the evidence is generally low. Further well-designed longitudinal studies are warranted on specific classes of pesticides and on possible interactions between different types of compounds.

Endocrine Disrupting Compounds Removal Methods from Wastewater in the United Kingdom: A Review

Endocrine disrupting compounds (EDCs) are contaminants with estrogenic or androgenic activity that negatively impact human and animal communities. These compounds have become one of the most significant concerns for wastewater treatment in recent decades. Several studies have evaluated EDC removal methods from wastewater across the globe, including the United Kingdom (UK). Accordingly, the current study reviews EDC removal methods from municipal/domestic wastewater in the United Kingdom (UK) for the period of 2010–2017. The current study analysed original research articles (250), review articles (52), short communication (43), and other associated documents via the ScienceDirect.com database. A total of 25 published articles, which covered EDC removal methods from UK wastewaters, were reviewed rigorously. The research highlights that despite the relative efficacy of existing chemical and physical methods for removing certain EDCs from wastewater, there is emerging evidence supporting the need for more widespread application of nature-based and biological approaches, particularly the use of biofilms. The analysis reveals that there have been relatively few research studies on EDC removal methods carried out in the UK in the 2010–2017 period. Only four papers addressed the removal of specific endocrine disrupting compounds from UK municipal wastewater, and none of the studies addressed EDC removal by using direct biofilms. Finally, this review suggests that more research is needed to remove EDCs, particularly through the application of biofilms, from municipal wastewater in current scenarios.

A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models

In recent years, there has been an increasing demand for predictive and sensitive in vitro tools for drug discovery. Split complementation assays have the potential to enlarge the arsenal of in vitro tools for compound screening, with most of them relying on well-established reporter gene assays. In particular, ligand-induced complementation of split luciferases is emerging as a suitable approach for monitoring protein–protein interactions. We hereby report an intracellular nanosensor for the screening of compounds with androgenic activity based on a split NanoLuc reporter. We also confirm the suitability of using 3D spheroids of Human Embryonic Kidney (HEK-293) cells for upgrading the 2D cell-based assay. A limit of detection of 4 pM and a half maximal effective concentration (EC50) of 1.7 ± 0.3 nM were obtained for testosterone with HEK293 spheroids. This genetically encoded nanosensor also represents a new tool for real time imaging of the activation state of the androgen receptor, thus being suitable for analysing molecules with androgenic activity, including new drugs or endocrine disrupting molecules.