The Journal of Steroid Biochemistry and Molecular Biology

Background Isotretinoin (ISO) is a systemic retinoid widely used for the treatment of acne vulgaris, yet its potential effects on male reproductive function remain unclear with previous studies reporting inconsistent findings. Methods and finding We conducted a multi-model study combining data from the Danish Young Men Study (DYMS) cohort with ex vivo cultured human testis tissue and an in vitro Retinoic Acid Receptor alpha (RAR) reporter gene assay. In the DYMS cohort semen parameters and reproductive hormone concentrations assessed on a specific examination day were compared between ISO-users (n=322) and non-users (n=4,065). Ex vivo human testis tissue was exposed to ISO and RA for 96 hours to assess changes in hormone secretion and germ cell dynamics, while the in vitro RAR assay evaluated ISO for effects on RA-induced RAR-signaling. In the DYMS cohort, ISO use was associated with reduced FSH concentrations, higher inhibin B/FSH ratio, and elevated estradiol concentrations, along non-significant tendencies of lower sperm counts. These associations persisted even after cessation of ISO use but were not evident in men first initiating ISO use 1 year after their examination date. In the ex vivo testis tissue, ISO exposure altered hormone secretion, including increased testosterone, but did not affect germ cell proliferation or apoptosis. ISO also inhibited RA-induced RAR-signaling in vitro in a competitive manner. Conclusions These findings suggests that ISO-use may influence male reproductive function, potentially with persistent effects. Further randomized clinical trials are needed to validate these effects.

Autoimmune diseases have shown biased proportion in female population, existing clinical investigations of sex hormones in autoimmune populations have been relatively limited in terms of patient size and types of hormones investigated. In this study, we examined the relationship of sexual hormones and autoimmune antibodies in a large cohort of US women. This retrospective study sample included a total of 15319 female subjects medical information that were collected between December 2015 to May 2019 and tested in the Vibrant America Clinical Laboratory. The present serum sample was limited to female participants who had ever menstruated at the time of blood collection and completed the testing of the autoimmune antibodies and sex hormones. We focused on a total of 13 clinically significant autoantibodies including antinuclear antibody (ANA), 11 anti-extractable nuclear antigens (anti-ENAs), anti-cyclic citrullinated peptide 3 (anti-CCP3), and 11 female sex hormones. First, the prevalence of serological autoantibodies in a large set of adult female subjects divided by the menopause age was investigated. Next, the levels of sex hormones were compared in the seropositive autoimmune subjects and seronegative controls across the pre- and post-menopausal female groups. The presented study involving a large cohort of females showed no statistically different levels of sex hormones in seropositive autoimmune subjects and matched controls except for DHEA-s.

Objective Sodium valproate (SV) is a widely used anti-epileptic drug with well-established reproductive and teratogenic effects, yet its impact on the developing prepubertal reproductive system remains poorly understood. This study investigated the effects of prepubertal SV exposure on gonadal development, including folliculogenesis, testicular architecture and steroidogenic gene expression in male and female mice in vivo. Methods CD1 mouse pups received intraperitoneal injections of saline (control), low or high dose SV (50 or 100 mg/kg, respectively) on postnatal days (PND) 6, 8, and 10. On PND17 the animals were culled and gonads dissected. Ovaries and testes were assessed histologically using haematoxylin and eosin staining. Ovarian follicle number, stage and health were quantified. Testicular tubule morphology and key testicular cell numbers (germ, Sertoli, spermatogonial stem cells, and interstitial/Leydig cells) were evaluated using immunofluorescence with automated image analysis. Expression of key steroidogenic genes (CYP11A1, STAR, CYP19A1 in ovaries; INSL3, STAR, CYP11A1 in testes) was measured by RT-qPCR. Results SV exposure did not significantly affect ovarian follicle number, distribution or health. Testicular morphology and the density of Sertoli, spermatogonial stem cells, and interstitial cells were likewise unaffected. However, a dose-dependent trend toward reduced germ cell density was observed in males at a high dose SV, though this did not reach statistical significance (P = 0.058). Steroidogenic gene expression was unaffected by SV exposure in both ovaries and testes across all treatment groups. Significance This study provides the first in vivo assessment of short-term prepubertal SV exposure on gonadal development in both sexes. The overall findings suggest that brief SV exposure during the prepubertal window does not cause overt gonadotoxicity. Nonetheless, the trend toward reduced testicular germ cell density warrants further investigation with longer exposure durations and functional fertility endpoints, to better inform clinical risk assessment in paediatric patients receiving SV.

Background: Male hypogonadism is associated with sexual, physical, and psychological manifestations, and testosterone replacement therapy (TRT) is used to restore testosterone levels and relieve symptoms. Objective: To evaluate the efficacy of TRT in adult men with hypogonadism across erectile function, hypogonadal symptoms, fatigue, body composition, waist circumference, urinary symptoms, and quality of life. Methods: This systematic review and meta-analysis followed PRISMA guidance. PubMed/MEDLINE, Google Scholar, and ScienceDirect were searched. Fourteen studies involving 1,388 participants were included. Effect estimates were synthesized as standardized mean differences using random-effects models, and heterogeneity was assessed with I2, Cochrans Q, and t2. Results: TRT was associated with favorable pooled effects on erectile function, hypogonadal symptoms, fatigue, lean mass, fat mass, waist circumference, and quality of life. No consistent effect was observed for lower urinary tract symptoms. Heterogeneity was high for erectile function, hypogonadal symptoms, waist circumference, and urinary symptoms; moderate for body-composition outcomes; and low for fatigue and quality of life. Randomized trials generally yielded smaller effect estimates than observational studies. Conclusions: TRT was associated with improvement across several clinical and body-composition outcomes in hypogonadal men, although effect magnitude varied substantially by outcome and study design. The high heterogeneity for several outcomes and the larger estimates in observational studies warrant cautious interpretation and support the need for longer, high-quality randomized trials.

Background and AimsPregnancy requires dynamic, stage-specific adaptations in maternal liver metabolism and growth to sustain fetal development while preserving systemic homeostasis. Estrogen signaling, which significantly increases during pregnancy, is primarily mediated in hepatocytes by estrogen receptor (ER). Although hepatic ER regulates female liver metabolism under non-pregnant conditions, its role in pregnancy-induced hepatic remodeling remains unclear. MethodsWe studied non-pregnant and pregnant control and liver-specific ER knockout (LERKO) mice across gestational stages using longitudinal physiological measurements, liver transcriptomics, targeted metabolomics, histological assessment of cell proliferation, and metabolic phenotyping. ResultsIn control mice, pregnancy elicited sequential hepatic remodeling characterized by early induction of cell-cycle programs, a mid-gestational peak in hepatocyte proliferation with transient suppression of selected metabolic pathways, and late reactivation of specific metabolic programs. Chronic hepatic ER deficiency alters this temporal pattern. LERKO livers showed premature activation of proliferative and anabolic transcriptional programs, changes in amino acid- and fatty acid-related metabolic pathways, and altered temporal regulation of AKT-mTORC1-related signaling. At mid-gestation, LERKO mice displayed reduced hepatocyte proliferation, altered expression of metabolic and insulin-related genes, blunted gestational glucose adaptation without overt evidence of systemic insulin resistance, and changes in the light/dark-phase metabolic patterns. ConclusionsThese findings suggest that hepatic ER is required for the appropriate stage-specific coupling of liver growth, metabolic remodeling, and insulin-responsive signaling during pregnancy. Its loss is associated with gestational hepatic maladaptation and systemic metabolic phenotypes, providing a framework for investigating estrogen-dependent mechanisms underlying pregnancy-associated metabolic and liver disorders. HighlightsHepatic ER is required for stage-specific liver remodeling during pregnancy. Loss of hepatic ER alters temporal coupling of liver growth and metabolism. LERKO mice show early changes in amino acid- and fatty acid-related pathways. Hepatic ER loss reduces proliferation and alters gestational glucose adaptation. Hepatic ER loss is associated with altered light/dark-phase metabolic organization. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC="FIGDIR/small/743939v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@d52bborg.highwire.dtl.DTLVardef@b27511org.highwire.dtl.DTLVardef@23b286org.highwire.dtl.DTLVardef@19d9314_HPS_FORMAT_FIGEXP M_FIG C_FIG

Peroxisome proliferator-activated receptor (PPAR) is a nuclear receptor that plays a central role in metabolic adaptation by regulating gene expression programs involved in lipid utilization and energy homeostasis. Although the hepatic functions of PPAR and its target genes are well characterized, less is known about its transcriptional activity in extrahepatic tissues. Pemafibrate (K-877), a selective PPAR modulator (SPPARM), is a high-affinity PPAR ligand with improved potency and selectivity compared with conventional fibrates. Here, we investigated pemafibrate-induced tissue-specific transcriptional responses comparing wild type and whole-body PPAR-deficient littermate mice. Twelve-week-old male PPAR+/+ and PPAR-/- mice were treated daily with pemafibrate (0.1 mg/kg) or vehicle by oral gavage for 14 days. We performed genome-wide transcriptomic analyses in the liver, jejunum, kidney, soleus muscle, and BAT. Pemafibrate-induced transcriptional changes were entirely dependent on PPAR, as no differentially expressed genes were detected in PPAR-/- mice. The magnitude and nature of the response were tissue-specific, with the strongest transcriptional effects observed in the liver and jejunum, whereas the kidney, soleus muscle, and BAT showed limited responses. In the liver, pemafibrate primarily induced the expression of genes involved in fatty acid metabolism, mitochondrial function, and energy production. In the jejunum, pemafibrate activated lipid metabolic programs together with tissue-specific pathways related to cellular homeostasis. Collectively, our findings show that pemafibrate is a highly selective PPAR activator with a preferential transcriptional activity in the liver and intestine after oral administration. This study provides a resource for defining tissue-specific PPAR-responsive gene networks and expands our understanding of PPAR functions beyond the liver.

Background: Accumulating evidence suggests that the prevalence of hypercortisolism in patients with cardiometabolic risk factors is much higher than previously thought. This study aimed to evaluate the prevalence of ACTH-independent hypercortisolism across the spectrum of cardiometabolic risk. Methods: Participants were prospectively recruited into three cohorts to undergo protocolized assessment of adrenal physiology:1) normotensive participants; 2) participants with hypertension and obesity, but without diabetes; 3) participants with diabetes and overweight/obesity. All participants (n=216) underwent overnight 1 mg dexamethasone suppression testing followed by cosyntropin stimulation test, and 24-hour urine free cortisol (UFC) testing. ACTH-independent hypercortisolism was defined as post-dexamethasone serum cortisol >1.8 g/dL (50 nmol/L) with a concomitant post-dexamethasone ACTH [&le;]10 pg/mL (2.2 pmol/L). Result: 16 of 216 (7.4%) participants were found to have ACTH-independent hypercortisolism, including 5.6% (4/71) in the normotensive cohort, 6.2% (5/81) in the hypertension-obesity cohort and 10.9% (7/64) in the diabetes-obesity cohort. Age, body mass index, hemoglobin A1c, blood pressure, renal function, morning ACTH, and 24-hour UFC were similar among those with and without ACTH-independent hypercortisolism. Following cosyntropin stimulation, those with ACTH-independent hypercortisolism had higher stimulated cortisol levels (greater ACTH-dependent responses) when compared to those without ACTH-independent hypercortisolism: 25.8 +/- 4.5 versus 20.6 +/- 4.1 g/dL (711 +/- 124 v. 568 +/- 113 nmol/L) (P<0.001). Conclusions: In this prospective study, the prevalence of ACTH-independent hypercortisolism paralleled the burden of cardiometabolic risk features. ACTH-independent hypercortisolism was associated with greater ACTH-stimulable cortisol production, identifying a unique biochemical phenotype. These findings reveal new insights into cortisol pathophysiology as a function of ACTH and cardiometabolic risk profiles.

Obesity is a major public health challenge of the 21st century, particularly in low- and middle-income populations. Adipogenesis plays a central role in the development of obesity and associated metabolic disorders, as it determines adipocyte number, size, and function. The 14-3-3 protein family comprises seven paralogs in mammals that regulate multiple cellular processes, yet their specific roles during adipogenesis remain poorly understood. In this study, we characterized the expression profiles of 14-3-3 paralogs during the early and late stages of adipogenic differentiation using quantitative PCR under standard adipogenic differentiation medium and modified drug-supplemented conditions. We found that the expression of specific paralogs is strongly influenced by the composition of the differentiation medium. The absence of insulin led to an early increase in Ywhaz, which could not be maintained during late adipogenesis and was associated with impaired adipogenic differentiation. In contrast, stimulation with incretins in combination with insulin induced late expression of Ywhag and Ywhab paralogs and promoted the formation of a greater number of smaller lipid droplets. These findings indicate that individual 14-3-3 paralogs exert distinct and context-dependent effects on adipogenesis, highlighting their potential roles as modulators of adipocyte differentiation and metabolic function.

Background: Endometriosis is an estrogen-driven, inflammatory disorder affecting ~10% of menstruators, causing severe pain. Current treatments that reduce estrogen or inflammation have inconsistent efficacy and poor tolerability. Therapies identified via drug-repositioning methods, which target patient-specific pathways, offer a promising alternative. Simvastatin and primaquine were identified as potential treatments, strongly reversing endometriosis-associated gene expression pathways, and behavioral testing in an animal model showed that these drugs diminish endometriosis-associated pain. This paper examines their effects in human assembloids to provide proof-of-concept validation and insights into their mechanisms of action relevant to endometriosis. Methods: We established human endometrial assembloids using immortalized epithelial (12Z) and stromal fibroblast (iEc) cell lines, as well as primary tissue from patients. Assembloids were generated in 96-well agarose molds with both cell types. We tested ibuprofen, fenoprofen, primaquine, and simvastatin. Following a 24-hour exposure, assembloids and monolayer cultures were harvested and stranded mRNA-seq libraries were sequenced on an Illumina NovaSeqX Plus System. Results: Gene set enrichment analysis showed key pathway reversals: Primaquine reversed the chemical carcinogenesis ROS pathway in assembloids. Simvastatin reversed cytokine-cytokine receptor interaction in both epithelial and stromal cell lines, and the cytoskeleton in muscle cells pathway in stromal cell lines. Fenoprofen reversed the calcium signaling pathway in stromal cell lines. Conclusion: Simvastatin induced the most significant gene expression changes, notably reversing cytokine-cytokine receptor interactions in both cell lines, mirroring findings from our rat model. Despite the small sample size limitation, these experiments highlight the promise of assembloid models to test therapeutic candidates for endometriosis.

Introduction: The evidence synthesis on literature characterizing changes in body fatness and adiposity distribution with sex steroid measurements in different matrices (blood, tissue, urine and other excreted matrices) is lacking. Methods: PubMed and Web of Science were used to identify eligible studies published until June 2026. Random-effects meta-analyses were conducted, and the strength of evidence was determined based on the Continuous Update Project criteria. Results: Of 16,875 screened references, 140 studies deemed relevant: 96 on circulating blood biomarkers from 39 weight loss intervention (surgical and lifestyle) and 60 observational studies reporting direct measure of adiposity distribution (e.g., DEXA, MRI), 20 studies on biomarkers measured in urine or other excreted matrices (i.e., saliva, urine, breast ductal lavage), and 25 tissue-based studies (e.g., adipose, breast, endometrial, skin). Weight loss was associated with a reduction of total and free testosterone, and an increase in sex hormone binding globulin (SHBG). While no significant associations were found between weight loss and premenopausal estradiol, dehydroepiandrosterone sulfate, and androstenedione. Decrease in total and free estradiol and estrone were observed in postmenopausal women. Findings in observational studies generally supported those results, as well as tissue-based studies on estrogen-related markers despite the heterogeneous tissue types. Studies on biomarker measured in excreted matrices were largely inconclusive. Conclusion: Strong evidence of a probable association was found between body fatness and SHBG, androgens, and postmenopausal estrogens. Limited evidence remained for other sex steroids.

Postmenopausal women are vulnerable to metabolic dysfunction. Compelling evidence suggests this is because they lack the protective effects of estrogens. We have shown that circulating estrogens regulate daily eating and activity rhythms in female mice and protect them from obesity and metabolic dysfunction. However, few studies have investigated whether postmenopausal women have disrupted eating and sleep-activity rhythms that could contribute to metabolic dysfunction. The purpose of this cross-sectional study was to investigate the relationship between sleep and eating rhythms and metabolic risk in postmenopausal women with overweight or obesity. Seven days of sleep and food intake behavior data were collected from postmenopausal women with overweight or obesity. Sleep timing was assessed using wrist-worn actigraphy and sleep logs. First and last calorie intake times were collected daily using text messaging. Visceral adipose tissue (DXA), insulin resistance (Matsuda index), blood pressure, and triglycerides were analyzed as markers of metabolic risk. In 122 postmenopausal women (mean {+/-} SD: 58.1 {+/-} 3.6 yr, 33.2 {+/-} 5.2 kg/m2 BMI), we found that later sleep onset was associated with greater visceral adipose tissue (p=0.0007), but not insulin resistance, blood pressure, or triglycerides. Next, we used a random forest model to quantify the relative contributions of sleep and meal timing variables to predict visceral adipose tissue. The top predictors of visceral adipose tissue were time of sleep onset, sleep duration, and time of last caloric intake. These data suggest that interventions that advance sleep onset may reduce metabolic risk in postmenopausal women.

Ketogenic (keto) diets have gained popularity due to their potential benefits on weight loss and metabolic conditions. However, the consequences of consuming a keto diet before, during, and after pregnancy on maternal and offspring health remain incompletely understood. To investigate this, female Swiss Webster mice were randomly assigned to either a keto or control diet. After 6 weeks on diet, females were mated and maintained on their respective diets throughout pregnancy and lactation. In experiment 1, glucose tolerance tests (GTT) were performed on pregnant dams at gestational day (GD) 16.5. At GD 17.5, dams were euthanized, urine was collected for ketone analysis, and fetal and placental weights were recorded. In experiment 2, dams delivered naturally and one male and one female offspring per dam were retained and weighed weekly. At 12 weeks of age, offspring underwent GTT, followed by euthanasia and serum collection for insulin and leptin analysis. Compared with controls, keto dams exhibited increased body weight, impaired glucose tolerance, elevated urinary ketones, and reduced circulating insulin during late gestation (p<0.05). Fetuses from keto dams were significantly smaller (p<0.05), whereas placental weight and placental efficiency were unchanged. Offspring from keto dams weighed less than controls during early postnatal development but demonstrated catch-up growth by 12 weeks of age. At 12 weeks, keto offspring exhibited a modest but statistically significant improvement in glucose tolerance compared to control offspring (p<0.05). Male keto offspring had significantly decreased serum insulin (p<0.05) compared to male control offspring, while no differences were observed in females. Serum leptin levels did not differ between diet groups, although sex differences present in control offspring were not observed in keto offspring. Together, these findings suggest that maternal ketogenic diet consumption is associated with altered maternal glucose homeostasis during pregnancy and reduced fetal growth in this mouse model. Although offspring exhibited only modest metabolic alterations during early adulthood, the observed changes in growth trajectory and sex-specific metabolic profiles support further investigation into the long-term consequences of maternal ketogenic diet exposure during pregnancy.

Several potentially potent anticancer drugs have been identified by in vitro evaluation, such as Andrographolide. These compounds show strong anticancer activity in vitro, but struggle to reach effective concentrations in the bloodstream when taken orally because they dissolve poorly in water or break down rapidly in the body. Bioenhancers, which are compounds that have potential to improve drug stability in the body, offer an alternative solution to overcome this limitation. Naringin and Quercetin have been identified as candidate bioenhancers, and have been hypothesized to potentially slow rapid first pass metabolism of poorly bioavailable drugs. Our work focuses on testing Naringin and Quercetin because they are flavonoids with therapeutic potential, due to their anti-inflammatory and antioxidant properties. Data from the hepatic microsomal assays performed on Naringin and Quercetin suggest moderate to proficient periods of stability in the body, with Naringin having 91.86% remaining, while Quercetin had 74.84% remaining. When administered alongside Andrographolide, a drug known to rapidly degrade in the body, Naringin raised its metabolic stability from 38.93% to 80.77% and on the other hand, Quercetin raised Andrographolide metabolic stability from 38.93% to 86.70%. In addition, plasma protein binding assays show the percentage of compounds available at the target site where Naringin was observed to be 49.32% bound and Quercetin found to be 50.14% bound, implying 50.68% of Naringin, and 49.86% of Quercetin available at the target site, respectively. This preliminary study explores whether Quercetin and Naringin could act as bioenhancers by remaining stable and available in plasma and by slowing the metabolism of poorly bioavailable drugs such as Andrographolide.

Background: Dietary sodium intake is a modifiable determinant of blood pressure, and impaired renal sodium handling is implicated in the pathogenesis of hypertension. Whether dietary sodium intake modulates renal tubular sodium transporter abundance among normotensive adults at risk for hypertension is unknown. Methods: Forty-six normotensive adults with normal kidney function at risk of developing hypertension were prospectively enrolled to complete a very low sodium (LS) diet followed by a high sodium (HS) diet. Urinary extracellular vesicle (uEV) proteins were quantified from a 24-hour urine collection at the end of each diet. We compared normalized uEV protein concentrations between diets and tested whether diet modified uEV protein associations with serum aldosterone, urinary aldosterone, and serum cortisol-cortisone ratio under either diet using Wilcoxon rank sums and linear mixed effects models, respectively. Results: Participants achieved median [IQR] urine sodium 13.2 [7.7-19.9] mEq/24h on the LS diet and 244.0 [197.1-297.0] mEq/24h on the HS diet. LS intake induced greater renin activity, greater plasma and urinary aldosterone, and lower urinary cortisol, when compared to HS intake. In parallel, NCC, ENaC, SGLT2, OXSR1, and Pendrin uEV concentrations were significantly higher during LS intake than HS intake (p<0.05 for all), whereas TRPV5 and AQP2 uEV concentrations were lower during LS intake than HS intake (p<0.001 for both). Conclusions: Controlled dietary sodium modulation induced nephron-segment specific changes in uEV protein abundance. These findings demonstrate the ability of uEVs to serve as a non-invasive tool to probe in vivo renal sodium, water, and electrolyte handling.

Polyendocrine Metabolic Ovarian Syndrome (PMOS, formerly PCOS) is associated with chronic low-grade inflammation. This study aimed to characterize the peripheral cytokine landscape in women with PMOS and identify cytokines associated with disease pathogenesis and phenotypic variation. Plasma concentrations of 48 cytokines were quantified using a multiplex bead-based assay in 102 women with PMOS and 114 healthy controls. We performed receiver operating characteristic (ROC) analysis, odds ratio estimation, correlation analysis, and network analyses to characterise cytokine alterations and identify phenotype-associated inflammatory signatures. Fourteen cytokines were significantly elevated in PMOS compared with healthy controls: IL-18, IL-1Ra, IL-12p70, IL-13, IL-16, MCP-1, MCP-3, CTACK, Eotaxin, RANTES, IP-10, M-CSF, HGF, and {beta}-NGF. ROC analysis identified IL-16, CTACK, and IP-10 as potential discriminators of phenotype A, whereas IL-18, M-CSF, and HGF demonstrated promising discriminatory performance for phenotype B. Odds ratio analysis identified 11 cytokines with odds ratios greater than 1, with IL-16 showing the strongest association with PMOS. Correlation analysis revealed 10 strongly correlated cytokine pairs (r [&ge;] 0.7) in the PMOS group, suggesting coordinated regulation of inflammatory mediators. Gene ontology and network analysis highlighted alterations in chemotaxis and immune cell migration pathways, implicating these processes in PMOS-associated immune dysregulation. These findings reveal a distinct peripheral inflammatory cytokine signature and phenotype-associated differences in PMOS. IL-16 emerged as a prominent candidate for further investigation, given its strong association with PMOS. Peripheral cytokine profiling may provide a minimally invasive approach to characterising immune alterations and identifying candidate biomarkers for disease characterisation and phenotype stratification.

Omega-3 polyunsaturated fatty acids (n-3 PUFA) influence white adipose tissue (WAT) lipid buffering capacity; however, sex-specific regulation remains understudied. We recently reported that male mice fed a diet containing high alpha-linolenic acid (ALA) had increased WAT mass, reduced serum triglycerides and elevated lipolysis compared to mice fed a diet containing recommended levels of ALA, independent of background dietary protein. The current study examined whether subcutaneous and visceral WAT (scWAT, vWAT) lipolytic activity was altered in female C57BL/6N mice (n=16/group) fed low-ALA (1% energy) or high-ALA (3% energy) diets containing skim milk protein (SMP) or soy protein isolate (SPI) for 8 weeks. Body weight, WAT depot weights and serum triglycerides were unchanged in response to ALA content. Lipolytic markers were mostly unchanged by ALA content except for an increase in adipose triglyceride lipase (ATGL) in vWAT, while diets containing SPI modestly reduced serum cholesterol levels and increased total hormone-sensitive lipase (HSL) content in vWAT. Collectively, WAT lipolytic markers in female mice showed minimal response to diets containing high ALA content, unlike that previously reported in male mice. These results highlight the importance of considering both sexes to ensure generalizability of findings when investigating diet regulation of WAT lipid metabolism.

The success of modern antiretroviral therapy (ART) has increased the life expectancy of people living with HIV to levels approaching that of uninfected individuals. For women living with HIV (WLWH), this means that more will survive to undergo menopause and experience the consequences of decreased ovarian hormone levels, particularly estrogen (E2). The recent change in federal guidance for use of postmenopausal hormone therapy is increasing demand for both E2-alone and E2+progestogen formulations to control adverse symptoms of menopause. The consequences and efficacy of hormone therapy in WLWH are thus an important issue for WLWH and their healthcare providers. The role of E2 replacement in postmenopausal WLWH is a significant issue because of its potential effects on control the viral reservoir and its demonstrated beneficial metabolic effects in uninfected postmenopausal women. To address these questions, we employed a novel nonhuman primate model of postmenopausal WLWH undergoing E2 replacement. Reproductively competent female rhesus macaques were infected with simian immunodeficiency virus (SIV) and then subjected to a daily ART regimen. After complete suppression of plasma viremia, all animals were ovariectomized (OVX) and then implanted with Silastic capsules containing either cholesterol vehicle or sufficient E2 to restore pre-OVX plasma levels. Plasma and cell-associated viral dynamics, immune responses, body composition, systemic and tissue-specific metabolic parameters, cytokine profiles, and parameters of bone health were followed longitudinally from baseline through 34 weeks of E2 deficiency or replacement. We found that E2 status did not significantly affect plasma or tissue viral dynamics or overall metabolic homeostasis. However, E2 replacement exerted beneficial effects on several aspects of bone health in spite of a chronic inflammatory state that persisted following effective ART suppression of the SIV reservoir. Our findings suggest that hormone therapy, specifically E2 replacement, offers benefit to WLWH, particularly with respect to bone loss.

Background/Objectives: Aldehyde dehydrogenase 2 (ALDH2), a mitochondrial enzyme, mitigates cellular stress by detoxifying reactive aldehydes produced during alcohol exposure and endogenous metabolic processes. ALDH2*2 polymorphism, which results in deficient catalytic activity, is prevalent in East Asian populations and is associated with an increased risk of cancer and other diseases. While the effects of ALDH2 deficiency on cellular stress responses are well documented, the specific metabolic dysregulations arising from this deficiency require further investigation. Methods; An ALDH2 knockout subline of SKBR-3 cells was established using CRISPR/Cas9 strategy. The metabolomic profiles of control (SKBR-3/C) and ALDH2 knockout (SKBR-3/ADKO) cells were evaluated via ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Results: ALDH2 knockout in SKBR-3 cells resulted in increased cellular stress, evidenced by elevated levels of 8-OHdG, reactive oxygen species (ROS), and malondialdehyde (MDA). Metabolomic analysis of the paired cell lines assigned a total of 5,033 signals, of which 281 were significantly different (fold change [&ge;] 2, p-value <0.05) between the two groups. Pathway analysis with MetaboAnalyst identified significant metabolic pathways associated with ALDH2 deficiency, particularly those involving arginine biosynthesis, and arginine and proline metabolism. ALDH2 deficiency reprogramed arginine metabolism toward polyamine pathways while suppressing the urea cycle. Network analysis further revealed alterations in amino acid and lipid metabolism in ALDH2-deficient cells. Conclusions; This study provides a novel insight into the role of ALDH2 in cancer metabolism and may have broader implication for cancer biology.

AimThe aim of the present study was to determine whether pharmacological inhibition of aldosterone synthesis during the stress-hyporesponsive period (SHRP) affects behaviour and adrenocortical stress responsiveness later in development and whether these effects differ between males and females. MethodsNewborn Wistar rat pups (males n=40, females n=40) were treated with aldosterone synthase inhibitor FAD286 (30 mg/kg per day, orally) or vehicle from PND3 to PND9. To verify the pharmacodynamic action of FAD286, serum and adrenal glands from 10-day-old pups were analysed. The remaining pups were weaned on PND21 and underwent open-field (PND23), elevated plus-maze (PND29) and salt-preference testing. At PND46, half of each group was exposed to restraint stress for 120 min. ResultsIn 10-day-old pups, treatment with FAD286 resulted in increased gene expression of CYP11B2 (aldosterone synthase) and CYP11B1 (11-beta-hydroxylase) in the adrenal glands, increased serum levels of corticosterone, and decreased concentrations of serum aldosterone. FAD286 did not modify the general locomotor activity assessed in juvenile rats. Inhibition of aldosterone synthase by FAD286 resulted in altered anxiety-like behaviour in a sex-dependent manner. Postnatal FAD286 treatment led to increased anxiety-like behaviour in female, but not male rats. During adolescence, early FAD286 treatment increased overall aldosterone concentrations without altering the aldosterone response to restraint. Basal corticosterone concentrations were unchanged, whereas the response to restraint was enhanced. ConclusionsThe present study demonstrates that transient inhibition of aldosterone synthesis during the SHRP led to alterations in anxiety-related behaviour and adrenocortical regulation later in development, with some behavioural effects being sex-dependent.

Uterine leiomyomas, commonly known as fibroids, are benign tumors of the female reproductive tract. They are the most common tumor of the female reproductive system and the leading cause of hysterectomy in the United States. Progesterone, estrogen, and their receptors are important drivers for the development and growth of leiomyomas. How estrogen drives leiomyoma growth remains unclear. We have previously shown that simvastatin can inhibit leiomyoma growth by modulating intracellular calcium. This suggests that calcium signaling is important for leiomyoma growth. We therefore hypothesized that estrogen may drive leiomyoma growth by stimulating calcium release through the membrane estrogen receptor GPR30 (also known as GPER). We found that estrogen dose-dependently induced calcium release in leiomyoma cells. The GPR30-specific agonist G-1 also induced calcium release, indicating that GPR30 mediates estrogen-mediated calcium release. The GPR30 antagonist G-15 blocked estrogen-induced calcium release and significantly reduced leiomyoma proliferation. Our results demonstrate that GPR30 is a major driver of estrogen-dependent growth in leiomyoma cells by modulating intracellular calcium. These findings may have high therapeutic relevance to leiomyomas and other estrogen-dependent tumors.