Classical Steroid Research Articles

2α-Methyl-5α-androstan-17β-ol-3-one-17β-heptanoate

Drostanolone is a popular synthetic dihydrotestosterone derivative and an anabolic–androgenic agent which belongs to the steroid class. The crystal structure of a new polymorph of the esterified prodrug of drostanolone, namely drostanolone enanthate, was elucidated using single crystal X-ray diffraction. Furthermore, it was analyzed using the thermal DTA/TGA technique and FT-IR spectroscopy.

Comprehensive targeted profiling of multiple steroid classes in rodent plasma using liquid chromatography-mass spectrometry

BackgroundReliable quantification of multiple steroid classes in biological fluids within a single method remains an analytical challenge despite many previously published methods. Crosstalk of positional isomers, overlap of stereoisomer fragmentation patterns, differing proton affinities, in-source fragmentation, varying stability of protonated ions in the gas phase across steroid classes, and non-existence of steroid-free matrix are the main challenges limiting the number of simultaneously profiled steroids. ResultsIn this study, we focused on the development of a derivatization-free, achiral, high-throughput, and cost-effective UHPLC-MS/MS approach that allows simultaneous profiling of a spectrum of 38 steroids covering progestogens, androgens, corticosteroids, and estrogens, while properly addressing the hurdles of steroid analysis. Within a 20-min method, 16 stereoisomers and 15 positional isomers were fully resolved within a single run while separated from 7 additional non-interfering steroids and matrix interferences in rodent plasma. Protein precipitation (PP) and supported liquid extraction (SLE) methods using only 40 μL of sample were developed to achieve the lowest possible limits of quantification. Nevertheless, 5α-dihydroprogesterone and 3α,5α-THDOC could be only qualitatively assessed when using PP. In contrast, DHEA-S could not be quantified or identified when using SLE. A novel surrogate matrix-background subtraction approach, using rat plasma after the animal's adrenalectomy, has been implemented into the optimized PP-UHPLC-MS/MS workflow, successfully validated according to the unified ICH/EMA M10 guidelines, and compared to the traditional quantification strategies. Moreover, the validity of the newly adopted approach has been verified by the targeted profiling of multiple biologically active endogenous steroids in more than 500 samples of mouse plasma in total. SignificanceUnderestimation of hurdles associated with steroid analysis often compromises the accurate steroid quantification. Our comprehensive, fully validated UHPLC-MS/MS method targeting a wide spectrum of endogenous steroids, mitigating steroid crosstalk and using a minimal sample volume together with a novel surrogate matrix-background subtraction approach significantly advances steroid analysis for research and clinical applications covering multiple biological scopes.

Patterns of Corticosteroid Prescriptions among Patients at Community Pharmacies in Lahore, Pakistan

Background: Corticosteroids are a class of steroid hormones produced by the adrenal gland and are used to treat many diseases, including autoimmune diseases and inflammatory diseases. Besides their role in many diseases, long-term use, misuse, and overuse cause side effects including hypertension, diabetes mellitus, and osteoporosis. Objective: The aim of current study is to evaluate the prescription pattern of corticosteroids in different community pharmacies, among patients, in Lahore. Methods: The cross-sectional stratified convenient study performed using a validated questionnaire to analyse the prescription pattern of corticosteroids in patients at different community pharmacies in Lahore. Total 92 patients are participating in this study. Results: A total of 92 patients take part in the study. In this research 67.4% females are participated. The average age of patients is above 30 years. Asthma is the most common reason for corticosteroids administration (19.6%) followed by rheumatoid arthritis (12.0%) whereas in inflammatory bowel disease they are least prescribed (1.1%). Prednisolone is the most prescribed drug (33.7%) and Oral route is the frequently given route. Conclusion: Our data shows that the prescription pattern of corticosteroid must be improved. Health care providers should pay attention to counseling of patients and guide them about dose tapering to avoid adverse effects.

Metabolit Sekunder dan Uji Bioaktivitas Sitotoksik Ekstrak Buah Sumpit (Brucea javanica (L.) Merr) terhadap Artemia salina Leach

Research carried out with the title of Secondary Metabolite Identification and bioactivity Fruit Extract Sumpit (Brucea javanica (L) Merr) to the shrimp larvae Artemia salina Leach. This study aims to identifying classes of secondary metabolites as well as test the bioactivity of secondary metabolites sumpit. Identifikasi fruit extracts performed to test the Chopsticks fruit extracts and fractions against reagent, while the bioactivity test was conducted using Brine Shrimp Lethality Test (BSLT). Data were analyzed using the method of Reed and Muench to get Lethality Concrentation value 50% (LC50). The results showed that the fruit extract Sumpit contain the alkaloid class of secondary metabolites, karatenoid, saponins, tannins, phenolic compounds, steroids, and terpenoids. Sumpit fruit bioactivity test results had LC50 values of the methanol extract of 3.10 ppm, n-hexane fraction of 2.09 ppm, 0.79 ppm ethyl acetate fraction and n-butanol fractions of 0.74 ppm.

Identification of bile acids in snake bile by hydrogen/deuterium exchange mass spectrometry and quantitative structure-retention relationship analysis

Natural bile acids, a class of steroids with a valeric acid side chain at the C-17 position, present significant challenges in separation and analysis due to structural similarities, isomerism, and large polarity differences. Therefore, advanced analytical methods are essential for the accurate identification and quantification of bile acids. This study conducted a comprehensive qualitative analysis of bile acids by integrating liquid chromatography-tandem mass spectrometry (LC-MS/MS), hydrogen-deuterium exchange tandem mass spectrometry (HDX-MS/MS), and quantitative structure-retention relationship (QSRR) methods. Firstly, LC-MS/MS conditions were optimized to enhance chromatographic separation and improve the reliability of characteristic fragment ions. MS/MS fragmentation rules for bile acids were derived from the mass spectral data of bile acid standards and validated through HDX-MS/MS experiments. Secondly, potential bile acids in snake bile were identified based on these validated fragmentation rules, and a QSRR model was established to predict the retention times of the proposed structures. Thirdly, HDX-MS/MS was applied to assist in identifying bile acid isomers. Finally, a total of 150 bile acids, including 11 free bile acids (free BA), 5 glyco-bile acids (GBA) and 134 tauro-bile acids (TBA), were detected in snake bile. Thirteen bile acids were accurately characterized by comparing their retention time and MS/MS spectra with standards. Forty-nine bile acids were reasonably annotated using the QSRR model and HDX-MS/MS. This study is notable for being the first to utilize the QSRR and HDX-MS/MS techniques for the annotation of bile acids in snake bile, providing a robust framework for the structural elucidation of these compounds.

Comsol Multiphysics modeling of an electrochemical biosensor using carbon nanotubes for detecting urinary estrogen receptor

A class of steroid hormones known as estrogens is essential for the health of the heart, bones, and reproductive system. Changes in estrogen levels have been connected to several health problems, such as endocrine disorders, metabolic syndromes, and cancer. In pharmaceutical applications, environmental monitoring, and medical diagnostics, biosensors that measure estrogen levels are essential. This study models estrogen detection biosensors based on urine liquid, horseradish peroxidase biorecognition, and carbon nanotubes (CNT) using Comsol Multiphysics. This study demonstrates that most interactions happen at the upper boundary of the concave pillars put inside the box. Besides, it shows that the velocity has the highest value between the concave pillars inside the box. The results demonstrate that the number of interactions (absorption and adsorption) rises with increasing the concave pillars' area, affecting the biosensor output.

Lack of membrane sex steroid receptors for mediating rapid endocrine responses in molluscan nervous systems.

Despite the lack of endogenous synthesis and relevant nuclear receptors, several papers have been published over the decades claiming that the physiology of mollusks is affected by natural and synthetic sex steroids. With scant evidence for the existence of functional steroid nuclear receptors in mollusks, some scientists have speculated that the effects of steroids might be mediated via membrane receptors (i.e. via non-genomic/non-classical actions) -a mechanism that has been well-characterized in vertebrates. However, no study has yet investigated the ligand-binding ability of such receptor candidates in mollusks. The aim of the present study was to further trace the evolution of the endocrine system by investigating the presence of functional membrane sex steroid receptors in a mollusk, the great pond snail (Lymnaea stagnalis). We detected sequences homologous to the known vertebrate membrane sex steroid receptors in the Lymnaea transcriptome and genome data: G protein-coupled estrogen receptor-1 (GPER1); membrane progestin receptors (mPRs); G protein-coupled receptor family C group 6 member A (GPRC6A); and Zrt- and Irt-like protein 9 (ZIP9). Sequence analyses, including conserved domain analysis, phylogenetics, and transmembrane domain prediction, indicated that the mPR and ZIP9 candidates appeared to be homologs, while the GPER1 and GPRC6A candidates seemed to be non-orthologous receptors. All candidates transiently transfected into HEK293MSR cells were found to be localized at the plasma membrane, confirming that they function as membrane receptors. However, the signaling assays revealed that none of the candidates interacted with the main vertebrate steroid ligands. Our findings strongly suggest that functional membrane sex steroid receptors which would be homologous to the vertebrate ones are not present in Lymnaea. Although further experiments are required on other molluscan model species as well, we propose that both classical and non-classical sex steroid signaling for endocrine responses are specific to chordates, confirming that molluscan and vertebrate endocrine systems are fundamentally different.

Mechanisms of Cancerprotective Effects of Phytosterols. Literature Review

Cancer is recognized as the second leading cause of mortality in the Russian Federation. Prolonging the life of oncology patients involves treatment with toxic drugs, causing multiple side effects. Today, scientists around the world are striving to find non-toxic drugs. The present study explores phytosterols. Phytosterols refer to a class of steroids widely distributed in plants as an essential component of plant cell membranes. They include stigmasterol, beta-sitosterol, and campesterol. Stigmasterol has been found to increase the expression of proapoptotic genes (Bax, p53) and decrease the expression of the antiapoptotic gene Bcl-2 in HepG2 liver cancer cells. Stigmasterol is able to induce cell arrest in G0-G1 phase (stationary phase), resulting in fewer cells in the G2/M phase (division phase). It induces apoptosis and protective autophagy in gastric cancer cells while inhibiting the Akt/mTOR signaling pathway. β-sitosterol exhibits growth inhibitory and cytotoxic effects against a number of established cancer cell lines in vitro and in vivo, and remains free from acute/subacute toxic effects. β-sitosterol is widely used to treat chronic prostate diseases. In 2020, spendings on dietary supplements rich in beta-sitosterol accounted for $24 827 065 in the USA. Campesterol induces cell apoptosis via the mitochondrial pathway. It appears cytotoxic to U937 hepatocellular cancer cells. Campesterol induces cell apoptosis and activates proapoptotic signaling in ovarian cancer cell lines of a person. The present literature review demonstrates that specific substances in this group, beta-sitosterol, stigmasterol, and campesterol, provide pronounced antitumor effects.

Structure and the Anticancer Activity of Vitamin D Receptor Agonists.

Vitamin D is a group of seco-steroidal fat-soluble compounds. The two basic forms, vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol), do not have biological activity. They are converted in the body by a two-step enzymatic hydroxylation into biologically active forms, 1α,25-dihydroxyvitamin D2 [ercalcitriol, 1,25(OH)2D2] and 1α,25-dihydroxyvitamin D3 [calcitriol, 1,25(OH)2D3], which act as classical steroid hormones. 1,25(OH)2D3 exerts most of its physiological functions by binding to the nuclear vitamin D receptor (VDR), which is present in most body tissues to provide support to a broad range of physiological processes. Vitamin D-liganded VDR controls the expression of many genes. High levels of 1,25(OH)2D3 cause an increase in calcium in the blood, which can lead to harmful hypercalcemia. Several analogs of 1,25(OH)2D3 and 1,25(OH)2D2 have been designed and synthesized with the aim of developing compounds that have a specific therapeutic function, for example, with potent anticancer activity and a reduced toxic calcemic effect. Particular structural modifications to vitamin D analogs have led to increased anticancer activity and reduced calcemic action with the prospect of extending work to provide future innovative therapies.

DNA methylation-mediated 11βHSD2 downregulation drives the increases in angiotensin-converting enzyme and angiotensin II within preeclamptic placentas.

Preeclampsia (PE) is a complex human-specific complication frequently associated with placental pathology. The local renin-angiotensin system (RAS) in the human placenta, which plays a crucial role in regulating placental function, has been extensively documented. Glucocorticoids (GCs) are a class of steroid hormones. PE cases often have abnormalities in GCs levels and placental GCs barrier. Despite extensive speculation, there is currently no robust evidence indicating that GCs regulate placental RAS. This study aims to investigate these potential relationships. Plasma and placental samples were collected from both normal and PE pregnancies. The levels of angiotensin-converting enzyme (ACE), angiotensin II (Ang II), cortisol, and 11β-hydroxysteroid dehydrogenases (11βHSD) were analyzed. In PE placentas, cortisol, ACE, and Ang II levels were elevated, while 11βHSD2 expression was reduced. Interestingly, a positive correlation was observed between ACE and cortisol levels in the placenta. A significant inverse correlation was found between the methylation statuses within the 11βHSD2 gene promoter and its expression, meanwhile, 11βHSD2 expression was negatively correlated with cortisol and ACE levels. Invitro experiments using placental trophoblast cells confirmed that active GCs can stimulate ACE transcription and expression through the GR pathway. Furthermore, 11βHSD2 knockdown could enhance this activating effect. An invivo study using a rat model of intrauterine GCs overexposure during mid-to-late gestation suggested that excess GCs in utero lead to increased ACE and Ang II levels in the placenta. Collectively, this study provides the first evidence of the relationships between 11βHSD2 expression, GCs barrier, ACE, and Ang II levels in the placenta. It not only contributes to understanding the pathological features of the placental GCs barrier and RAS under PE conditions, also provides important information for revealing the pathological mechanism of PE.

The translational aspects of glucocorticoid biorhythmicity in modern therapeutics

Glucocorticoids are a class of steroid hormones, vital for mammalian life. They have a plethora of biological effects, mainly supporting metabolic, cognitive, and immunological functions. The pharmacological use of glucocorticoids makes them one of the most frequently prescribed drugs across all continents, and in all types of forms. Nevertheless, a number of serious adverse effects accompany the prolonged treatment with high doses of glucocorticoids. Research developments over the last 20 years have gradually reshaped the way we think about glucocorticoid-based therapeu¬tics. Aside their circadian rhythm and their delayed regulatory influence over an extensive number of sensitive genes, glucocorticoids also possess an underlying, ultradian rhythm, and also exert rapid, non-genomic effects. The notion that chronicity of glucocorticoid stimulation may differentially modulate the type of biological effects of the hormone brings various chronopharmacological concepts on the table of modern glucocorticoid-based therapeutics.

Cytochrome P450 Mining for Bufadienolide Diversification.

Bufadienolides are a class of steroids with a distinctive α-pyrone ring at C17, mostly produced by toads and consisting of over 100 orthologues. They exhibit potent cardiotonic and antitumor activities and are active ingredients of the traditional Chinese medicine Chansu and Cinobufacini. Direct extraction from toads is costly, and chemical synthesis is difficult, limiting the accessibility of active bufadienolides with diverse modifications and trace content. In this work, based on the transcriptome and genome analyses, using a yeast-based screening platform, we obtained eight cytochrome P450 (CYP) enzymes from toads, which catalyze the hydroxylation of bufalin and resibufogenin at different sites. Moreover, a reported fungal CYP enzyme Sth10 was found functioning in the modification of bufalin and resibufogenin at multiple sites. A total of 15 bufadienolides were produced and structurally identified, of which six were first discovered. All of the compounds were effective in inhibiting the proliferation of tumor cells, especially 19-hydroxy-bufalin (2) and 1β-hydroxy-bufalin (3), which were generated from bufalin hydroxylation catalyzed by CYP46A35. The catalytic efficiency of CYP46A35 was improved about six times and its substrate diversity was expanded to progesterone and testosterone, the common precursors for steroid drugs, achieving their efficient and site-specific hydroxylation. These findings elucidate the key modification process in the synthesis of bufadienolides by toads and provide an effective way for the synthesis of unavailable bufadienolides with site-specific modification and active potentials.

The impact of estradiol on serotonin, glutamate, and dopamine systems.

Estradiol, the most potent and prevalent member of the estrogen class of steroid hormones and is expressed in both sexes. Functioning as a neuroactive steroid, it plays a crucial role in modulating neurotransmitter systems affecting neuronal circuits and brain functions including learning and memory, reward and sexual behaviors. These neurotransmitter systems encompass the serotonergic, dopaminergic, and glutamatergic signaling pathways. Consequently, this review examines the pivotal role of estradiol and its receptors in the regulation of these neurotransmitter systems in the brain. Through a comprehensive analysis of current literature, we investigate the multifaceted effects of estradiol on key neurotransmitter signaling systems, namely serotonin, dopamine, and glutamate. Findings from rodent models illuminate the impact of hormone manipulations, such as gonadectomy, on the regulation of neuronal brain circuits, providing valuable insights into the connection between hormonal fluctuations and neurotransmitter regulation. Estradiol exerts its effects by binding to three estrogen receptors: estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), and G protein-coupled receptor (GPER). Thus, this review explores the promising outcomes observed with estradiol and estrogen receptor agonists administration in both gonadectomized and/or genetically knockout rodents, suggesting potential therapeutic avenues. Despite limited human studies on this topic, the findings underscore the significance of translational research in bridging the gap between preclinical findings and clinical applications. This approach offers valuable insights into the complex relationship between estradiol and neurotransmitter systems. The integration of evidence from neurotransmitter systems and receptor-specific effects not only enhances our understanding of the neurobiological basis of physiological brain functioning but also provides a comprehensive framework for the understanding of possible pathophysiological mechanisms resulting to disease states. By unraveling the complexities of estradiol's impact on neurotransmitter regulation, this review contributes to advancing the field and lays the groundwork for future research aimed at refining understanding of the relationship between estradiol and neuronal circuits as well as their involvement in brain disorders.

Phytoconstituents of Leaves and Roots Ethanolic Extract of Talinum paniculatum and Their Biological Activities

Javanese ginseng (Talinum paniculatum) is known to the people of Indonesia to have properties in medicine and food. People's culture generally uses leaves and roots that, until now, constituent information has yet to bereported. Therefore, variations of phytoconstituents in the leaves and roots of T. paniculatum were investigated using Gas Chromatography-Mass Spectrometry (GC-MS). The roots of T. paniculatum contain 17 active compounds dominated by N,N'-Dibutyl-N,N'-dimethylurea. The leaf has only 16 active compounds, but Python compounds from the diterpenoids class dominate it. In general, T. paniculatum contains a variety of steroid class compounds, namely Stigmasterol; Stigmast-5-en-3-ol; and 9,19-Cycloergost-24(28)-en-3-ol, 4,14-dimethyl-, acetate, (3,β., 4.α., 5.α.) - in the leaves, and stigmasterol and .γ.Sitosterol in the root. Python is known to have high efficacy as an antimicrobial, antifungal, antibacterial, antiparasitic, antimutagenic, and antioxidant. At the same time, steroid compounds are anti-cancer, antioxidant, anti-tumor, antidiabetic, and anti-inflammatory agents. The results of identifying compounds in T. paniculatum can be used as a reference in optimizing the use of T. paniculatumin the future.

Innovative bioremediation of dexamethasone in aquatic ecosystems using Rhodococcus sp. D32: Pathway discovery and reduction of ecotoxicity

In response to the widespread environmental concern of endocrine disruptors, our research investigated the aerobic biodegradation of dexamethasone (DEX), a widely used synthetic glucocorticoid with endocrine-disrupting properties, by strain Rhodococcus sp. D32. This innovative strain showcases remarkable DEX-degrading capabilities, with the half-life for degrading 500 μg/L DEX in MH broth being just 7.56 h. Utilizing high-resolution mass spectrometry, we identified six transformation products (TPs) and revealed a unique steroid nucleus-opening pathway marked by A-ring lactonization and subsequent lactone hydrolysis. This newly discovered pathway deviates from the classical steroid 9,10-seco pathways and previous transformations that preserved the intact steroid nucleus, thus expanding our knowledge of microbial steroid transformations. Additionally, the degradation process progressively converts the parent DEX into less ecotoxic products, significantly reducing the overall ecotoxicity of the system. This reduction remains evident when considering the combined toxicity of DEX and its TPs, indicating a promising approach for lessening environmental harm. Strain D32 also proved effective in various authentic aquatic environments, including fishpond water, urban sewage, river water, and reservoir water, where its introduction markedly expedited the dissipation of DEX. Our findings underscore the potential of strain D32 as a powerful tool for the bioremediation of DEX, presenting an efficient and eco-friendly strategy to alleviate the effects of this endocrine disruptor in aquatic ecosystems.

The Role of Endogenous Brassinosteroids in the Mechanisms Regulating Plant Reactions to Various Abiotic Stresses

Plants are vulnerable to many abiotic stresses, resulting in reduced plant productivity. Its adaptation to unfavorable environments relies on transmitting external stress signals into internal signaling pathways. A series of stress response mechanisms have been developed. Among them, brassinosteroids (BRs) are a class of steroid hormones that are widely involved in plant growth, development, and stress response. Via genetics, proteomics, and genomics studies, the major components of signaling and signaling pathways through a series of phosphorylation cascade reactions have been identified in model plants such as Arabidopsis. Numerous studies have shown that BRs play important roles in plant responses to drought, temperature, salt, heavy metals, and other environmental stresses. The application of BRs to improve plant stress resistance has become the focus of research in recent years, especially the regulation of stress via endogenous BRs. Therefore, this paper systematically summarizes the research progress related to endogenous BR levels and provides an overview of BR biosynthesis and the signaling pathway, as well as the function of endogenous BRs in the response to abiotic stresses.

Bioinspired Skeletal Reorganization Approach for the Synthesis of Steroid Natural Products.

ConspectusSteroids, termed "keys to life" by Rupert Witzmann, have a wide variety of biological activities, including anti-inflammatory, antishock, immunosuppressive, stress-response-enhancing, and antifertility activities, and steroid research has made great contributions to drug discovery and development. According to a chart compiled by the Njardarson group at the University of Arizona, 15 of the top 200 small-molecule drugs (by retail sales in 2022) are steroid-related compounds. Therefore, synthetic and medicinal chemists have long pursued the chemical synthesis of steroid natural products (SNPs) with diverse architectures, and vital progress has been achieved, especially in the twentieth century. In fact, several chemists have been rewarded with a Nobel Prize for original contributions to the isolation of steroids, the elucidation of their structures and biosynthetic pathways, and their chemical synthesis. However, in contrast to classical steroids, which have a 6/6/6/5-tetracyclic framework, rearranged steroids (i.e., abeo-steroids and secosteroids), which are derived from classical steroids by reorganization of one or more C-C bonds of the tetracyclic skeleton, have started to gain attention from the synthetic community only in the last two decades. These unique rearranged steroids have complex frameworks with high oxidation states, are rich in stereogenic centers, and have attractive biological activities, rendering them popular yet formidable synthetic targets.Our group has a strong interest in the efficient synthesis of SNPs and, drawing inspiration from nature, we have found that bioinspired skeletal reorganization (BSR) is an efficient strategy for synthesizing challenging rearranged steroids. Using this strategy, we recently achieved concise syntheses of five different kinds of SNPs (cyclocitrinols, propindilactone G, bufospirostenin A, pinnigorgiol B, and sarocladione) with considerably rearranged skeletons; our work also enabled us to reassign the originally proposed structure of sarocladione. In this Account, we summarize the proposed biosyntheses of these SNPs and describe our BSR approach for the rapid construction of their core frameworks. In the work described herein, information gleaned from the proposed biosyntheses allowed us to develop routes for chemical synthesis. However, in several cases, the synthetic precursors that we used for our BSR approach differed substantially from the intermediates in the proposed biosyntheses, indicating the considerable challenges we encountered during this synthetic campaign. It is worth mentioning that during our pursuit of concise and scalable syntheses of these natural products, we developed two methods for accessing synthetically challenging targets: a method for rapid construction of bridged-ring molecules by means of point-to-planar chirality transfer and a method for efficient construction of macrocyclic molecules via a novel ruthenium-catalyzed endoperoxide fragmentation. Our syntheses vividly demonstrate that consideration of natural product biosynthesis can greatly facilitate chemical synthesis, and we expect that the BSR approach will find additional applications in the efficient syntheses of other structurally complex steroid and terpenoid natural products.

METABOLOMICS OF LONGEVITY AND LIFESPAN

Abstract Serum metabolomics has been an important source of biomarkers of aging and longevity for years. This symposium will bring together investigators from large studies of human longevity to provide an overview of recent discoveries on serum metabolomics of aging and extreme human longevity, their connections to genetic variations, and highlight the challenges of correlating metabolomic profiles of aging in human studies and across multiple species. Dr. Sebastiani will describe results from analyses of serum metabolomics of participants enrolled in the Long Life Family Study, highlight similarities and differences between metabolomic profiles of old age and extreme old age, and some connections with genetics of extreme human longevity. Dr. Rappaport will connect specific variations of the APOE alleles to metabolomic profiles and describe a possible role of bioenergetics pathways in mediating the effect of APOE to longevity and resistance to Alzheimer’s disease. Dr. Monti will expand the characterization of metabolomics of aging and extreme human longevity in a large metabolomic study of very old centenarians by using traditional statistical analyses and novel machine learning techniques. His analysis identifies rich signatures of aging and longevity that include well known metabolites and point to bile acids and several classes of steroids as important marker of longevity. Analytical innovations will be taken further by Dr. Schork who will introduce a novel approach based on distance of profiles to analyze multiple metabolites simultaneously and show the value of this approach to analyze metabolomic profiles of maximum lifespan across multiple species. This is a Geroscience Interest Group Sponsored Symposium.

SERUM METABOLOMICS SIGNATURES OF EXTREME OLD AGE AND LONGEVITY

Abstract A total of 1,495 chemicals including 1,213 compounds of known identity and 282 compounds of unknown structural identity were profiled in serum samples collected at enrollment at Metabolon, Inc. from the blood serum of 213 subjects, including centenarians (n=80), offspring (n=70), and controls (n=63), mean ages of 105, 70, and 70, respectively, enrolled in the New England Centenarian Study (NECS). We performed metabolite- and metabolite module-based regression analyses on age, differential analyses comparing centenarian to offspring and control, and Cox proportional hazard-based survival analyses. We annotated the derived signatures by enrichment analysis using metabolite sets curated by Metabolon, RefMet, and Pathbank, among others. We identified 323 (436) metabolites that change with age at an FDR-corrected q-value of 0.01 (0.05), and 298 (407) metabolites differentially abundant between centenarians, their younger offspring, and controls. Our results confirm and expand upon previous metabolomics-based studies of aging, including decreased abundance with age of Tryptophan, DHEA-S, Pregnenolone, Glutathione, and Androgen, among others, and increased abundance of Amino acids (Tyrosines), Phenylacetylglutamine, Ornithine, Urea, Kynurenine, and Creatine, among others. To distinguish metabolites associated with longevity from those associated with extreme old age, we performed age-adjusted survival analysis, and rank-based enrichment analysis identified several metabolites significantly associated with survival independent of age, including secondary and primary (C 24) bile acids, and multiple classes of steroids (androgens, pregnenolone, progestin), among others. Additional analyses are ongoing to further annotate and characterize these findings. If validated, these results could contribute to the identification of novel healthy aging therapeutics.

Mitochondrial morphology in the mouse adrenal cortex: Influence of chronic psychosocial stress

Mitochondria within the adrenal cortex play a key role in synthesizing steroid hormones. The adrenal cortex is organized in three functionally specialized zones (glomerulosa, fasciculata, and reticularis) that produce different classes of steroid hormones in response to various stimuli, including psychosocial stress. Given that the functions and morphology of mitochondria are dynamically related and respond to stress, we applied transmission electron microscopy (TEM) to examine potential differences in mitochondrial morphology under basal and chronic psychosocial stress conditions. We used the chronic subordinate colony housing (CSC) paradigm, a murine model of chronic psychosocial stress. Our findings quantitatively define how mitochondrial morphology differs among each of the three adrenal cortex zones under basal conditions, and show that chronic psychosocial stress mainly affected mitochondria in the zona glomerulosa, shifting their morphology towards the more typical glucocorticoid-producing zona fasciculata mitochondrial phenotype. Analysis of adrenocortical lipid droplets that provide cholesterol for steroidogenesis showed that chronic psychosocial stress altered lipid droplet diameter, without affecting droplet number or inter-organellar mitochondria-lipid droplet interactions. Together, our findings support the hypothesis that each adrenal cortex layer is characterized by morphologically distinct mitochondria and that this adrenal zone-specific mitochondrial morphology is sensitive to environmental stimuli, including chronic psychosocial stressors. Further research is needed to define the role of these stress-induced changes in mitochondrial morphology, particularly in the zona glomerulosa, on stress resilience and related behaviors.