Steroid Concentrations Research Articles

Urinary haloacetic acid concentrations in relation to sex and thyroid hormones among reproductive-aged men

Sex and thyroid hormones are critical for male reproductive health. However, the associations between haloacetic acid (HAA) exposure – a known endocrine disruptor – and sex and thyroid hormones in humans remains unclear. We thus recruited 502 male participants seeking fertility evaluation from a reproductive center. We measured concentrations of sex and thyroid hormones in a single blood sample and dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) in repeated urine samples. Multivariable linear regression models were constructed to evaluate the associations between HAA concentrations and hormone measurements. After adjusting for potential confounders and urinary creatinine concentrations, urinary concentrations of TCAA were inversely associated with serum levels of sex hormone-binding globulin (SHBG), testosterone (T), T/luteinizing hormone ratio (T/LH), and thyroid stimulating hormone (TSH) (all P for trend < 0.10). Compared with participants in the lowest quartile of TCAA concentrations, those in the highest quartile had reduced serum levels of SHGB by 14.2 % (95% CI: −26.7, −3.0 %), T by 11.1 % (95% CI: −21.7, −1.3 %), T/LH by 21.0 % (95% CI: −36.7, −7.1 %), and TSH by 19.1 % (95% CI: −39.7, −1.5 %). Additionally, we observed inverse associations between continuous measurements of urinary HAAs and serum levels of free T, bioactive T, and estradiol. Our findings suggest that male HAA exposure may be associated with disrupted sex and thyroid function.

Phytochemical Screening of Ethanol Extract of Tarap Fruit Seeds (Artocarpus odoratissimus)

Tarap fruit (Artocarpus odoratissimus), also known as "terap", is a type of fruit that grows in various tropical regions in Southeast Asia. because almost every part of the fruit has pharmacological properties, it is often used as an ingredient in traditional medicine. The plant has been part of many different cultures for a long time, used as a food source, and is widely used to help improve health. This study aims to conduct phytochemical screening on tarap fruit seed extract to identify the secondary metabolite compounds contained in it with a focus on compounds such as polyphenols, alkaloids, tannins, saponins, steroids, and terpenoids. This study uses a qualitative experimental method to analyze phytochemistry by color test using various reagents from tarap fruit seed extract, to identify secondary metabolite compounds in the plant. Tarap fruit seed samples were obtained from local fruit sellers in Tarakan. Extraction of tarap fruit seeds was carried out using 96% ethanol with maceration technique, then examined the content of polyphenols, alkaloids, tannins, saponins, steroids and terpenoids. The results showed the presence of several secondary metabolites in tarap fruit seed extract, proving the content of polyphenols, alkaloids, saponins, and terpenoids, while tannins and steroids could not be detected. The conclusion is that the ethanol extract of tarap fruit seeds (Artocarpus odoratissimus) contains polyphenols, alkaloids, saponins, and terpenoids.

The effect of bisphenols on sex and thyroid hormone concentrations in cord blood among newborns

This study aims to investigate the associations of bisphenols with sex and thyroid hormones in cord blood among newborns. Four bisphenols, three hormones related to gonadal function, and four parameters related to thyroid function were measured in umbilical cord blood in 378 mother-newborn pairs. Multivariable linear regression, quantile-based g-computation (QGC), and Bayesian kernel machine regression were used. In the multivariable linear regression, bisphenol A (BPA) was associated with increased testosterone (TT) (regression coefficient, β = 0.049, 95% confidence interval, CI: 0.013,0.085; p = 0.007) and free tri-iodothyronine (FT3) levels (β = 0.019, 95% CI: 0.003, 0.035; p = 0.023), and decreased thyroid peroxidase antibody (TPOAb) (β = −0.053, 95% CI: 0.098, −0.008; p = 0.021). Consistently associations were observed in males, except TT, which was observed in females, and bisphenol AF (BPAF) was associated with decreased follicle-stimulating hormone (FSH) in females. These associations were also observed in a mixture of bisphenols. Moreover, we observed maternal prepregnancy body mass index (BMI) and delivery mode disparity in the relationship between bisphenols and sex and thyroid hormones. This study suggests that bisphenols may exert effects on sex and thyroid hormones in newborns, the effect may vary with sex differences, maternal prepregnancy BMI, and delivery mode.

Impact of benzo[a]pyrene, PCB153 and sex hormones on human ESC-Derived thyroid follicles using single cell transcriptomics

IntroductionEndocrine disruptors are compounds of manmade origin able to interfere with the endocrine system and constitute an important environmental concern. Indeed, detrimental effects on thyroid physiology and functioning have been described. Differences exist in the susceptibility of human sexes to the incidence of thyroid disorders, like autoimmune diseases or cancer. MethodsTo study how different hormonal environments impact the thyroid response to endocrine disruptors, we exposed human embryonic stem cell-derived thyroid organoids to physiological concentrations of sex hormones resembling the serum levels of human females post-ovulation or males of reproductive age for three days. Afterwards, we added 10 µM benzo[a]pyrene or PCB153 for 24 h and analyzed the transcriptome changes via single-cell RNA sequencing with differential gene expression and gene ontology analysis. ResultsThe sex hormones receptors genes AR, ESR1, ESR2 and PGR were expressed at low levels. Among the thyroid markers, only TG resulted downregulated by benzo[a]pyrene or benzo[a]pyrene with the “male” hormones mix. Both hormone mixtures and benzo[a]pyrene alone upregulated ribosomal genes and genes involved in oxidative phosphorylation, while their combination decreased the expression compared to benzo[a]pyrene alone. The “male” mix and benzo[a]pyrene, alone or in combination, upregulated genes involved in lipid transport and metabolism (APOA1, APOC3, APOA4, FABP1, FABP2, FABP6). The combination of “male” hormones and benzo[a]pyrene induced also genes involved in inflammation and NFkB targets. Benzo[a]pyrene upregulated CYP1A1, CYP1B1 and NQO1 irrespective of the hormonal context. The induction was stronger in the “female” mix. Benzo[a]pyrene alone upregulated genes involved in cell cycle regulation, response to reactive oxygen species and apoptosis. PCB153 had a modest effect in presence of “male” hormones, while we did not observe any changes with the “female” mix. ConclusionThis work shows how single cell transcriptomics can be applied to selectively study the in vitro effects of endocrine disrupters and their interaction with different hormonal contexts.

Phytochemical Analysis and Antibacterial Property of Green Okra Fruit Extract Against Enterococcus Faecalis Bacteria Tooth Root Canals

Background: Root canal treatment aims to eliminate bacteria that cause pulp abnormalities. Enterococcus faecalis is a bacteria in the infected tooth pulp, even those that have been treated. Irrigation was carried out together with biomechanical preparation using a combination of NaOCl and EDTA. The material is antibacterial, dissolves the organic and inorganic tissue of the root canal dentin, lacks odor and irritates the soft tissue. An alternative irrigation material from Jember cultivation is green okra. The fruit contains flavonoids, tannins, steroids, terpenoids. This study aims to analyze the phytochemical and antimicrobial properties of green okra fruit extract on the growth of E. Faecalis bacteria. Method: Phytochemical analysis to see the content of alkaloids, flavonoids, tannins, saponins, steroids and terpenoids. Antimicrobial analysis included the inhibition test using disc diffusion, MIC and MBC tests using the dilution method. Samples were green okra fruit extract 100%, 50%, 25%, 12.5%, 6.25%, 3.125%, 1.563%, 2.5% NaOCl, 17% EDTA. The MIC and MBC tests were examined for Optical Density values ​​with a spectrophotometer and then subcultured to see whether there was growth of bacterial colonies. Analysis of the research data used ANOVA and LSD parametric statistical tests Result: Phytochemical content sequentially is Polyphenols, Alkaloids, Flavonoids, Saponins, Tannins, Steroids, Terpenoids. Antimicrobial properties Green okra fruit extract concentration of 6.25% had the smallest inhibition zone, the largest OD value and the highest colony count. The concentration of 100% has the largest inhibition zone, the lowest OD, there are no bacterial colonies. Antimicrobial power The concentration of 100% is close to that of 17% EDTA and 2.5% NaOCl. In the Kruskal Wallis test, there were differences in the Mann-Whitney test study group, there were significant differences between all groups Conclusion: The phytochemical content of green okra fruit extract has antimicrobial activity against E. faecalis approaching 2.5% NaOCl and 17% EDTA at 100% concentration, 6.25% MIC concentration and 12.5% ​​MBC concentration. Keywords: Phytochemical, Antibacterial, Green okra fruit extract, Enterococcus faecalis

Hair steroid before and after COVID-19 in preschoolers: the moderation of family characteristics

BackgroundFrequent or prolonged exposure to stressors may jeopardize young children’s health. The onset of the COVID-19 pandemic, coupled with disruptions in daily routines and social isolation resulting from public health preventive measures, have raised concerns about its potential impact on children’ experienced stress, particularly for young children and vulnerable families. However, whether the pandemic was accompanied by changes in physiological stress remains unknown as perceived stress is not a good proxy of physiological stress. This study examined if preschoolers showed increasing hair steroid concentrations following the onset of the COVID-19 pandemic and whether family characteristics may have exacerbated or buffered these changes. Methods136 preschoolers (2–4 years) provided hair for steroid measurement (cortisol, dehydroepiandrosterone (DHEA), cortisone, cortisol-to-DHEA ratio, cortisol-to-cortisone ratio) in October-November 2019 (T0) and in July-August 2020 (T1). A 2-centimeter hair segment was analyzed, reflecting steroid production over the two months leading up to collection. Family income, conflict resolution and lack of cohesion, as well as parents’ COVID-19 stress were reported by parents. Linear mixed models for repeated measures and Bayes factors were used. ResultsNo significant changes were noted from before to after the onset of the COVID-19 pandemic for most hair steroids. However, a moderating role of family conflict resolution was noted. Children living with parents with a better ability to resolve conflicts had lower levels of DHEA compared to those who had more difficulty managing conflicts. Additionally, lower levels of family cohesion and income were linked to some steroids, especially DHEA, suggesting that these factors may relate to children’s physiological stress. Finally, boys had higher DHEA levels than girls. ConclusionOur findings suggest that stress biomarkers were comparable from before to during the COVID-19 pandemic. This observation holds true despite the pandemic being perceived by many as a novel, unpredictable, and potentially threatening event. Findings further suggest that family characteristics are associated with hair steroid, especially DHEA, which deserves further investigation.

Abstract PO4-27-04: GRACE-trial: A randomized active-controlled trial for vulvovaginal atrophy in breast cancer patients on endocrine therapy

Abstract Background: Breast cancer is the most common cancer type in women worldwide. Due to hormone receptor positivity in the majority of the breast cancer tumors is endocrine therapy a crucial part in the treatment landscape of breast cancer. Selective estrogen-receptor modulators or aromatase inhibitors are the used treatment options for endocrine therapy. These medicines generate a hypoestrogenic environment by reducing circulating estrogen or by altering the effect of estrogen on tissue cells by receptor blockade. As a common side effect, vulvovaginal atrophy occurs in a majority of breast cancer patients using endocrine therapy. Vulvovaginal atrophy has a significant impact on physical and psychological wellbeing due to negative influence on quality-of-life, self-esteem and sexuality. As a consequence, adherence of endocrine therapy for the standard duration of 5 to 10 years is challenging, resulting in higher rates of therapy interruption, leading to poorer prognosis with shorter distant disease free survival. The standard treatment for vulvovaginal atrophy in postmenopausal women is based on the use of local hormonal treatment. However, when a patient has a history of breast cancer, delay of treatment and undertreatment are ubiquitous. Methods &amp; design: In this first ever prospective randomized trial breast cancer patients on endocrine therapy with vulvovaginal atrophy will be treated with the available local treatment modalities with a 1:1:1:1 randomization: estrogen, dehydroepiandrosterone, moisturizers and a co-treatment of estrogen and probiotics. Patient-reported outcomes measurements will be implemented to investigate the efficacy of the implemented treatments. These outcome measures include symptom evaluation, impact on quality-of-life and sexuality. Safety of the treatments will be evaluated by assessing systemic sex hormones concentrations. Previous research indicated no increased recurrence risk, yet no randomized trials comparing the different modalities have been published. Primary objectives: Two primary objectives have been determined. Firstly, the efficacy of the different treatment modalities will be assessed. The assessment will be based on patient-reported outcome measurements (PROMs) and clinical evaluations such as vaginal pH and the vaginal maturation index, the latter a direct microscopic evaluation of the vaginal epithelium. Secondly, safety of the different groups will be assessed. Evaluation will be done be measuring sex hormone concentrations systemically, which will be a surrogate for safety evaluation. Secondary objectives: The secondary objective in this study is the identification of microbial alterations after treatment initiation. Identification of these alterations can help in further understanding of the pathophysiology of vulvovaginal atrophy and potentially create opportunities for new treatment strategies towards vulvovaginal atrophy in breast cancer patients on endocrine therapy. Registration: The current study is registered at the European Union Clinical Trials Register (EudraCT number 2021-001921-31). Trial status: Recruitment for the study was commenced in May 2022. At time of manuscript submission, patient recruitment was ongoing. Citation Format: Glenn Vergauwen, Piet Cools, Hannelore Denys, Tom Fiers, Koen van de Vijver, Liv Veldeman, Hans Verstraelen. GRACE-trial: A randomized active-controlled trial for vulvovaginal atrophy in breast cancer patients on endocrine therapy [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-27-04.

Abstract PS07-01: Exemestane and breast cancer prevention: how low can we go? Drug and biomarker tissue levels in a randomized presurgical trial on exemestane alternative dosing regimen

Abstract Exemestane is an effective drug to reduce breast cancer risk reaching an overall 65% reduction in breast cancer in the placebo–controlled phase III MAP.3 trial. To improve its acceptability in primary prevention programs, we are seeking the minimal effective dose. In a 3-arm presurgical trial of 4-6 weeks before breast surgery in 180 postmenopausal women with ER-positive breast cancer, we investigated the activity of alternative exemestane schedules: 25 mg per day (QD), 25 mg three times/week (TIW) or 25 mg per week (QW) and showed that in adherent participants TIW was not inferior to QD in reducing circulating estradiol (Serrano et al JAMA Oncol. doi:10.1001/jamaoncol.2023.0089). Moreover, Ki67 reduction was seen in all arms with no significant difference among arms. Here, we analyzed the concentration of sex steroids, exemestane, and its main metabolite in the cancer and adjacent non-cancerous breast tissue. Tissues samples were homogenized before liquid-liquid extraction. After reconstitution, samples were analyzed by coupling liquid chromatography with tandem mass spectrometry (Sciex QTRAP 6500, Nexera system, Shimadzu). We obtained breast cancer tissue from 93 and non-cancerous breast tissue from 117 participants to measure exemestane, 17-OH-exemestane, and sex steroids. Exemestane and 17-OH-exemestane concentrations were detectable only in the QD arm, while in TIW and QW arms levels were below the Lower Limit of Detection (&amp;lt; LLD). Median exemestane level was 3807 fmol/g and 17485 fmol/g and median 17-OH-exemestane level was 338 fmol/g and 1343 fmol/g in cancer and non-cancerous tissue, respectively. Interestingly, drug and its metabolite accumulated 4-5-fold in non-cancerous tissue compared to cancer tissue in the QD arm. Despite the between-arm drug concentration difference, estradiol was almost completely suppressed in all arms in the non-cancerous tissue, attaining level &amp;lt; LLD in QD and TIW arms, and barely detectable in QW arm. The median in the QW arm was &amp;lt; LLD (&amp;lt; LLD, interquartile range &amp;lt; LLD, 25.5 fmol/g) showing no differences in QD vs TIW and QD vs QW (p = 0.364 and p = 0.693 respectively). While a dose-response trend was observed in cancer tissue, estradiol level was &amp;lt; LLD (&amp;lt; LLD,52.2 fmol/g) on QD, 17.1 (&amp;lt; LLD, 125.3) on TIW, and 128 (&amp;lt; LLD, 224.8) on QW (p=0.046 QD vs TIW arms). Estrone showed a clear dose response trend among arms, whereas no differences were observed for testosterone and androstenedione for both cancer and non-cancerous tissue in all arms. The Ki-67 change was analyzed in the previous paper; here we report the data for those patients who had drug and hormones tissue concentration measured, where Ki-67 decreased in all arms: median Ki67 change from baseline was QD -8 (-10, -3), TIW -6 (-11, -2), QW -4 (-8, -1). Conclusions: Exemestane 25 mg three times a week maintains comparable activity to the standard dose on tissue estradiol suppression and Ki67 decrease. Considering the estradiol suppression in non-cancerous tissue of the lowest exemestane dose, QW might even be considered for breast cancer risk reduction in primary prevention. Further analyses are ongoing to investigate the correlation with other biomarkers including the role of polymorphic UGT2B17 genotype that could identify candidates to lower exemestane dosage. Citation Format: Davide Serrano, Harriet Johansson, Bjørn-Erik Bertelsen, Gunnar Mellgren, Parijatham Thomas, Katherine Crew, Nagi B Kumar, Debora Macis, Valentina Aristarco, Aliana Guerrieri Gonzaga, Sara Gandini, Mauro D’Amico, Tania Buttiron Webber, Irene Maria Briata, Stefano Spinaci, Viviana Galimberti, Giuseppe Viale, Lana A. Vornik, Eduardo Villar-Sanchez, Powel Brown, Brandy M Heckman-Stoddard, Eva Szabo, Bernardo Bonanni, Andrea De Censi. Exemestane and breast cancer prevention: how low can we go? Drug and biomarker tissue levels in a randomized presurgical trial on exemestane alternative dosing regimen [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PS07-01.

Lived Experiences of Sexuality and Sexual Functioning in Males with SCI: A Mixed-Methods Study.

Sexual dysfunction is highly prevalent in males with spinal cord injury (SCI) and has been recognized to be a key recovery priority. This cross-sectional, mixed-methods study aimed to investigate the major themes linked to sexual functioning in males with chronic (>1 year) SCI. Twenty male participants with SCI, aged 25 to 59 years, completed validated questionnaires exploring sexual function/satisfaction and health-related quality of life and a semi-structured interview with an experienced sexual medicine physician. Sex hormone concentrations and metabolic biomarkers, along with body composition and habitual physical activity levels, were assessed. Interview recordings were transcribed and thematic analysis performed using combined COM-B (Capability, Opportunity, Motivation, and Behavior) and biopsychosocial models to identify and organize major contributors and barriers to sexual functioning. Metabolic and hormonal biomarkers largely fell within normal physiological ranges despite reduced sexual functioning reported in our cohort (19/20 participants reported some degree of erectile dysfunction). Qualitative analysis of interview transcripts revealed 24 themes. Adaptability was important for improving sexual satisfaction. Attraction and attentiveness to sex and partners remained stable over time, while the desire for intimacy increased post injury. Sexual social norms, and comparisons to the able-bodied population, provided challenges for sexual activity and partnership. Environmental concerns regarding access to sexual health resources and accessible physical spaces during intimacy were relevant. Mood disorders and general life stressors negatively impacted sexual desire, while physical activity encouraged sexual activity. By considering a holistic view of sexuality in males with SCI, we identified key contributors and barriers to sexual functioning for the cohort studied.

The Effectiveness of Aqueous Extract of Ginger on Some Sex Hormones and Biochemical Parameters in Female Laboratory Mice (Mus Musculus L.)

Objective: Ginger is a flowering tropical plant that commonly used as a spice and flavoring having various pharmacological properties, several phytochemical compounds which has biological activity. The main bioactive compound in ginger is gingerol, have a powerful anti-inflammatory effect will greatly affect on fertility. Also, it has an effective role in many laboratory experiments to study the physiological activities of ginger extract on these functions. Aim: This study aimed to investigate the effects of ginger extract on some sex hormones like estrogen, progesterone, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in female mice, also its effect on total protein, albumin and globulin Methods: Thirty adult female mice of (Mus Musculus L.) weight (22-24g) were derived into three groups one control and two treated. Ginger water extract be ready to prepare with affecting doses according to experimental test on mice (3.5 and 7 mg body weight/mice). The concentration of serum female sex hormones (FSH, LH, progesterone and estrogen) were measured. Results: Ginger water extract caused a significant increase in the level of estrogen in both doses and significant increase in the level of FSH and LH in high dose only (P less than 0.01). While progesterone level show no significant changes. On the other hand, the serum total protein, albumin and globulin shows a significant increase only in total protein and globulin levels (P less than 0.01) in turn albumin level show no significant changes in comparison to control. Conclusion: The results of this study, ginger extract could increase the level of female sex hormones (estrogen, FSH and LH) and conceder as adjunctive natural therapy for improvement of female sex hormones as well as lead to increase in total protein and globulin level which is essential for boosting the immunity.

Renal Function and Transporters in the Na/K-ATPase α1S/S Mouse Model of High Affnity for Cardiotonic Steroids

Genetic ablation of ATP1A1 in the mouse renal proximal tubule (RPT) revealed that Na/K-ATPase (NKA) α1 tonically inhibits sodium and water reabsorption. Mechanistically, findings in KO mouse RPT and renal epithelial cells were consistent with a prevalent role of a NKA/Src-dependent downregulation of Na+/H+ exchanger 3 (NHE3). Pharmacologically, functional activation of this pathway has been obtained in vitro and ex vivo using low concentrations of NKA archetypal ligands cardiotonic steroids (CTS). However, the physiological implication of an activation of the endogenous CTS (eCTS)/NKA/Src on renal transporters including NHE3 has not been explored. We assessed the renal impact of CTS/NKA α1 receptor activation by comparing mice expressing the naturally resistant NKA α1 and age-matched mice genetically engineered to express a mutated NKA α1 with high affnity for CTS (R111Q/D122N double substitution). Mice obtained from established colonies at the University of Cincinnati were a gift from Dr. Jerry Lingrel. CTS-sensitive (α1S/S) male mice aged 3-6 months on a C57Bl6 background and age-matched controls (α1R/R) were used. Mice were born with a normal Mendelian ratio, with no observable difference between α1S/S and α1R/R. Renal Na/K-ATPase inhibitory dose-response curve to the CTS ouabain was shifted to the left, as expected, and occurred without detectable change of total renal NKA activity, NKA α1 or β1 protein contents. Metabolic cage studies revealed that increased α1 sensitivity to CTS resulted in a significant increase in urine output in α1S/S mice compared to age-matched α1R/R controls (3.9±0.3 vs. 2.4±0.2 ml/24h; n=6, p&lt;0.01). Consistent with an activation of the NKA/Src receptor by eCTS, western blot analyses of renal cortical preparation revealed an increased phosphorylation of NKA α1 at Y260 (Src target residue) and decreased NHE3 content (43.6 %; n=5, p&lt;0.01). RNA-seq analysis of whole kidneys from α1S/S and α1R/R littermate mice using the Illumina NovaSeq 6000 system (Novogene) and Volcano Plot (DEG data using Python) further showed differential expression of 296 genes (147 upregulated and 149 downregulated). Gene Set Enrichment Analysis (GSEA) using the WebGestalt toolkit showed that the altered pathways were mostly involved in solute transport. Taken together, these results provide genetic evidence of the physiological role of eCTS/NKAα1 on renal function in the intact mouse. Increased affnity to eCTS is suffcient to lead to an activation of NKAα1/Src, a downregulation of NHE3, and transcriptional regulation of renal transporters associated with increased urine output. This work was supported by AHA, Predoctoral fellowship #19PRE34450095, NIH DK129937, and NIH Grant P20GM103434 to the West Virginia IDeA Network of Biomedical Research Excellence. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Knock out of ELAVL1 affects steroid synthesis in adrenocortical carcinoma cell line NCI-H295R and leads to a less malignant phenotype

Introduction: Adrenocortical carcinoma (ACC) is a rare malignancy of the adrenal gland. With a 5-year-survival rate below 50% and no curative treatment options in advanced stages new therapeutic options in ACC are needed. Analyzing the ACC cohort of the cancer genome atlas we found a significant negative correlation of the expression level of a gene called Embryonic Lethal Abnormal Vision Like 1 (ELAVL1) and ACC patient survival. ELAVL1 has been described to be unfavorable in a lot of different tumor entities, but its function in ACC progression is still unknown. Methods: To evaluate the function of ELAVL1 in ACC progression we performed a CRISPR/Cas9 mediated knock out (KO) of the ELAVL1 gene in the ACC cell line NCI-H295R. The KO was confirmed on DNA level via Sanger sequencing and on protein level via immunofluorescence and western blot. Transcriptome changes were analyzed by next generation RNA sequencing (NGS). To characterize changes in cell function we performed soft agar colony forming assays and evaluated changes in steroid production via mass spectrometry. Results: Via CRISPR/Cas9 we knocked out ELAVL1 in NCI-H295R cells. NGS data showed 6926 significantly deregulated genes in ELAVL1-KO cells compared to control cells. There was an upregulation of 3468 genes, while 3458 genes showed a lower expression in ELAVL1 KO cells. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed a significant upregulation of 29 genes associated with aldosterone synthesis and a significant downregulation of 87 genes associated with pathways in cancer. Analyzing cell function via soft agar colony forming assays an impaired colony forming ability in ELAVL1 KO cells was seen. Furthermore, we could prove significant changes in steroid production. ELAVL1 KO led to an inhibition of androgen and glucocorticoid synthesis and an enhancement in mineralocorticoid synthesis. These changes could not only be proven by measuring steroid concentrations in cell culture supernatants, but also by NGS showing a downregulation of the genes CYP17A1 and CYP11B1 and an upregulation of CYP21A2 and CYP11B2. Conclusion: Our study showed that ELAVL1 affects the regulation of around 7000 genes in the ACC cell line NCI-H295R. Especially the inhibition of glucocorticoid synthesis shows that ELAVL1 may be a new target for therapeutic approaches in ACC treatment since hypercortisolism is one of the main symptoms in ACC and a negative prognostic factor for patient survival. The authors have nothing to disclose. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The Antityrosinase Activity of Mitragyna speciosa Korth Leaf Ethanolic Extract

The kratom plant (Mitragyna speciosa Korth) is a the plant that is traditionally used to soften the skin. Kratom leaves have very strong antioxidant activity so they are predicted also to have anti-tyrosinase activity. The potential inhibition of tyrosinase enzyme activity is thought to come from the content of flavonoid compounds, polyphenols, steroids, tannins, and quinones. This research aims to determine the tyrosinase inhibitory activity of ethanol extract of M. speciosa leaves. The inhibitory activity of the tyrosinase enzyme using L-DOPA as substrate and kojic acid as positive control. Antityrosinase testing was carried out by in vitro method using UV-vis spectrophotometer through determination of IC50 value. The results of the antityrosinase activity test of ethanol extract of M. speciosa leaves showed an IC50 value of 2117.708 ppm. The ethanol extract of M. speciosa leaves is scientifically proven to have antityrosinase activity with a weak category.

Comparison of assay methods for quantifying sex hormone concentrations across the menstrual cycle in rhesus macaques†.

Immunoassays have been the preferred method for steroid hormone analysis for more than 50years. Automated immunoassays (AIAs) offer high throughput, rapid data turnaround, and low cost for measuring steroid hormone concentrations. The application of liquid chromatography-tandem mass spectrometry (LC-MS/MS) for steroid quantification provides greater specificity and selectivity for individual steroids, the ability to simultaneously analyze multiple steroids, and high throughput and automation. We compared AIA and LC-MS/MS for analysis of 17beta-estradiol (E2) and progesterone (P4) over the course of several menstrual cycles in 12 rhesus macaques (Macaca mulatta). Serum samples were collected every 4 days across four menstrual cycles from each monkey. AIAs were performed on a Roche cobas e411 analyzer. LC-MS/MS analysis was performed on a Shimadzu-Nexera-LCMS-8060 instrument. Scatter plots with Passing-Bablok regression showed excellent agreement between AIA and LC-MS/MS for both E2 and P4. Bland-Altman plots revealed no bias for either method; however, AIA overestimated E2 at concentrations >140pg/ml and underestimated P4 at concentrations >4ng/ml compared to LC-MS/MS. A comparison of testosterone concentrations measured by AIA and LC-MS/MS in the same samples was also performed. In contrast to E2 and P4, AIA and LC-MS/MS yielded significantly different results for testosterone concentrations, with AIA consistently underestimating concentrations relative to those obtained by LC-MS/MS. Well-characterized automated immunoassays are an excellent tool for daily monitoring of monkey menstrual cycles or providing single data points requiring fast turnaround. In certain situations where AIAs may provide inaccurate estimations of E2 and P4 concentrations, LC-MS/MS assays are preferable.

Phytochemical screening and GC-FID identification of bioactive compounds in n-hexane, ethylacetate and methanol fractions of methanolic leaves extract of Azanza garckeana

Azanza garckeana is a tropical African plant commonly used for food and medicine. This study was aimed at evaluating the antioxidant, phytochemicals and the bioactive compounds present in n-hexane, ethylacetate and methanol fractions of the methanolic extract of Azanza garckeana leaves. Phytochemical screening and antioxidant assayed were carried out using standard methods, while bioactive compounds were analyzed using gas chromatography-flame ionization detector (GC-FID). The phytochemical compositions revealed considerable level of tannins and saponins with moderate concentration of flavonoids and steroids in the methanol fraction; whereas n-hexane fraction also showed considerable amount of tannins followed by saponins and a low concentration of steroids. However, ethylacetate fraction had the highest yield of tannins, steroids, and terpenoids which were significantly (p < 0.05) higher than methanolic fraction and n-hexane fraction respectively. The antioxidant activity of the methanolic crude extract of A. garckeana leaves showed that the % DPPH radical scavenging activity of the crude extract increased with decreased in concentration when compared with the standard antioxidant (ascorbic acid). The result of GC-FID analysis revealed the presence of nineteen constituents in n-hexane fraction, while ethylacetate and methanol fractions each have twenty constituents. This result showed that Azanza garckeana leaves contain health-promoting bioactive compounds with potent antioxidants.

Associations of pentachlorophenol exposure during pregnancy with maternal and infant reproductive hormones based on a birth cohort

Pentachlorophenol (PCP), a typical environmental endocrine disruptor and a new persistent organic pollutant, has been extensively used as a pesticide worldwide. Although its use has been restricted for decades, PCP remains prevalent in both the environment and human bodies. Despite the known endocrine-disrupting and exogenous hormonal effects of PCP, few epidemiological studies examined such impact, especially among sensitive populations and during critical periods. Based on a prospective birth cohort in Wuhan, China, we collected maternal (first trimester; 13.0 ± 1.02 gestational weeks) and infant urine samples (1.16 ± 0.22 months postpartum) from 720 mother-infant pairs. We aimed to examine the association of PCP exposure during early pregnancy with maternal and infant urinary sex steroid hormones, including estrogens (estrone, E1; estradiol, E2; estriol, E3), progestogens (progesterone, P4; pregnenolone, P5; 17α-OH-Progesterone, 17OHP4; 17α-OH-Pregnenolone, 17OHP5), and androgens (testosterone, Testo; dihydrotestosterone, DHT; dehydroepiandrosterone, DHEA; androstenedione, A4). Additionally, gonadotropins [follicle-stimulating hormone (FSH) and luteinizing hormone (LH)] were measured in infant urine. Detection frequencies of all the sex steroid hormones in the maternal urine samples (>99 %) were higher than those in the infants' [most ≥80 %, except for E1 (3.36 %) and E2 (21.4 %)]. Maternal urinary PCP concentration was found to be significantly related with increased maternal sex steroid hormone concentrations; each interquartile increase in PCP concentration was positively related with percent change of the hormones (%Δ) ranging from 26.6 % to 48.5 %. On the other hand, maternal PCP exposure was associated with significantly increased P4 in male infants [%Δ (95 % confidence interval): 10.5 (0.56, 21.4)] but slightly decreased P4 in female infants [−11.9 (−21.8, 0.68)]. In addition, maternal PCP exposure was significantly associated with decreased FSH [%Δ (95 % CI): −9.90 (−17.0, −2.18)] and LH [−8.44 (−16.0, −0.19)] in the female infants, but not in the male infants. Sensitivity analyses, excluding infertility related treatment, pregnancy complications, preterm birth, or low birth weight, showed generally consistent results. Our findings implied that maternal/prenatal PCP exposure might disrupt the homeostasis of maternal and infant reproductive hormones. However, further studies are needed to confirm the findings.

Canine and feline foetal fluids: Volume, hormonal and biochemical characterization during pregnancy.

This study aimed to evaluate the volume, the concentration of steroid hormones, and biochemical composition of the foetal fluids at different gestational ages in dogs and cats. Following the ovariohysterectomy, the allantoic and amniotic fluid samples were collected from pregnant bitches and queens and were assigned to different groups according to their gestational age. The canine and feline allantoic fluid volume increased during pregnancy, reached its maximum values on days 40-49 and then decreased. The canine and feline amniotic fluid volume increased steadily by the last days of pregnancy. In spite of significant changes of sex hormones in the foetal fluids, their concentration and ratios were not significantly different between male and female fetuses. The canine amniotic cortisol concentration increased until days 40-49 and decreased significantly afterwards. The maximum cortisol concentrations in the feline allantoic and amniotic fluids were observed on days 50-60 and 40-49, respectively. During the canine pregnancy, the concentrations of calcium, phosphorus, chloride, sodium, triglyceride, cholesterol, total protein, albumin and the activities of aminotransferase (AST), alkaline phosphatase (ALP), amylase and gamma-glutamyl transferase (GGT) in the amniotic fluid were higher than the allantoic fluid. The magnesium, potassium, lactate dehydrogenase (LDH) activity, creatine and lipase were higher in the allantoic fluid. In the feline allantoic fluid, potassium, magnesium, phosphorus, creatinine, albumin and glucose concentrations and the activities of creatine kinase (CK), GGT, LDH and lipase were higher. The ALP, AST activities, sodium and calcium concentrations were higher in the amniotic fluid (p<0.05). Volume of foetal fluids was determined in dogs and cats. Concentration of sex hormones did not different between male and female fetuses.

Disentangling the relationships of body mass index and circulating sex hormone concentrations in mammographic density using Mendelian randomization.

Mammographic density phenotypes, adjusted for age and body mass index (BMI), are strong predictors of breast cancer risk. BMI is associated with mammographic density measures, but the role of circulating sex hormone concentrations is less clear. We investigated the relationship between BMI, circulating sex hormone concentrations, and mammographic density phenotypes using Mendelian randomization (MR). We applied two-sample MR approaches to assess the association between genetically predicted circulating concentrations of sex hormones [estradiol, testosterone, sex hormone-binding globulin (SHBG)], BMI, and mammographic density phenotypes (dense and non-dense area). We created instrumental variables from large European ancestry-based genome-wide association studies and applied estimates to mammographic density phenotypes in up to 14,000 women of European ancestry. We performed analyses overall and by menopausal status. Genetically predicted BMI was positively associated with non-dense area (IVW: β = 1.79; 95% CI = 1.58, 2.00; p = 9.57 × 10-63) and inversely associated with dense area (IVW: β = - 0.37; 95% CI = - 0.51,- 0.23; p = 4.7 × 10-7). We observed weak evidence for an association of circulating sex hormone concentrations with mammographic density phenotypes, specifically inverse associations between genetically predicted testosterone concentration and dense area (β = - 0.22; 95% CI = - 0.38, - 0.053; p = 0.009) and between genetically predicted estradiol concentration and non-dense area (β = - 3.32; 95% CI = - 5.83, - 0.82; p = 0.009), although results were not consistent across a range of MR approaches. Our findings support a positive causal association between BMI and mammographic non-dense area and an inverse association between BMI and dense area. Evidence was weaker and inconsistent for a causal effect of circulating sex hormone concentrations on mammographic density phenotypes. Based on our findings, associations between circulating sex hormone concentrations and mammographic density phenotypes are weak at best.

Analisis Fitokimia Terhadap Tepung Bayam (Amaranthus Viridis)

Various plants can be developed into traditional medicines and raw materials for making healthy food products because they contain bioactive compounds in the form of potential secondary metabolites. One plant that contains various potential secondary metabolites and can be developed as a health product is spinach. Green spinach leaf powder is reported to contain alkaloids, flavonoids, saponins, tannins, anthraquinones, steroids, coumarins, carotenoids, and phenols. This research aims to analyze the content of tannins, alkaloids, saponins, triterpenes, steroids, and flavonoids. Spinach flour is obtained by drying spinach leaves using a dehydrator for ± 19 hours at a temperature of 60 oC. The dried spinach leaves are ground with a blender until smooth, then sieved with a 70-mesh sieve. Phytochemical analysis was carried out qualitatively using color reagents. The research results showed that spinach flour contains bioactive compounds, namely tannins, alkaloids, triterpenes, and flavonoids. Spinach flour has the potential to be used as a raw material in the development of functional food products that are rich in potential bioactive compounds to improve public health

Menstrual Cycle Phase Has No Influence on Performance-Determining Variables in Endurance-Trained Athletes: The FENDURA Project.

This study aimed to investigate the effect of the MC and endogenous sex hormone concentrations on performance-determining variables in three distinct MC phases in endurance-trained females. Twenty-one eumenorrheic trained/highly trained endurance athletes completed a standardized test battery during the early follicular phase (EFP), ovulatory phase (OP), and midluteal phase (MLP) for either one ( n = 7) or two test cycles ( n = 14). MC phases were determined using calendar-based counting, urinary ovulation testing, and verified with serum hormone analysis. MCs were retrospectively classified as eumenorrheic or disturbed. Disturbed MCs were excluded from analysis. The test battery consisted of 4-6 × 5-min submaximal stages with stepwise speed increases, a 30-s all-out double-poling ski ergometer test, and a maximal incremental treadmill running test. At a group level, there was no effect of MC phase or the serum concentrations of estrogen and progesterone on peak oxygen uptake (V̇O 2peak ), oxygen uptake at 4 mmol·L -1 blood lactate concentration, time to exhaustion, running economy, or mean 30-s power output (MPO 30s ). Serum testosterone concentration was positively associated with MPO 30s ( P = 0.016). Changes in V̇O 2peak from EFP to MLP were inconsistent between individuals and across cycles. None of the measured performance-determining variables were influenced by MC phase or serum estrogen or progesterone concentrations. Although some individual patterns could be observed, there was no indication that any single MC phase is consistently associated with improved or impaired V̇O 2peak on a group level.