Glucocorticoid Research Articles

The rational use of glucocorticoids may reduce the risk of readmission in menopausal women with knee osteoarthritis: results from a five-year longitudinal study

ObjectiveTo investigate the effect of glucocorticoids (GCs) on the risk of readmission in menopausal women with knee osteoarthritis (OA).Information and methodsThe study cohort comprised 80 menopausal women with knee OA treated at a tertiary hospital affiliated with Shanxi Medical University and who underwent follow-up between September 2018 and September 2023. Then the collected longitudinal monitoring data were used to construct a semi-variable coefficient shared Gamma frailty model (VCSGF). Based on the results of this model, we explored the impact of GCs on menopausal women with knee OA and made risk predictions.ResultsThe mean patient age at study entry was 64.7 ± 9.3 (range 50–82 years). And during the research, patients were admitted to the hospital a mean of 2.4 ± 1.8 times (range 1–11 times). Compared to patients who have not used the drug, the risk of early readmission in patients who have used GCs was reduced by 96% (HR = 0.04, 95%CI: 0.006 ~ 0.284, ). This protective effect diminished over time (). In addition, the risk of hospital admission with recurrent symptoms was roughly 3.35-fold higher in patients who drink alcohol than in patients who do not (95%CI: 1.661~6.794, ).ConclusionGC use reduced the risk of readmission in menopausal women with knee OA, although this effect diminished over time. Therefore, in patients without further contraindications, GCs may be used in moderation to reduce the readmission risk. In clinical practice, additional research is needed to investigate the timing and appropriate use of GC in the treatment of menopausal women with knee OA, and to develop a more rational program for GC use.

Squamous and basal skin cancers in 17,207 solid organ transplant recipients: Real-world data from national health insurance database in Poland.

Immunosuppressive therapy in organ transplant ensures proper graft function for many years, but it is burdened with a negative impact on the development of skin cancer in them. To characterize the impact of immunosuppressive therapy in transplant recipients on the development of non-melanoma skin cancers (NMSC). A total of 17,207 Polish patients who underwent liver, heart or kidney transplants between 2010 and 2022 and were on immunosuppression were included in the study. Immunosuppression was most commonly achieved using a regimen of tacrolimus (TAC) or cyclosporine A (CsA) combined with mycophenolic acid (MPA) and glucocorticosteroids (GS). Data on NMSC incidence from the National Health Fund in this population were analyzed and compared against incidence of NMSC in general Polish population in the same period. Renal transplant recipients demonstrated a significantly elevated risk of NMSC compared to the general population, with a 1-year cumulative incidence of 0.09% vs 0.04% (p < 0.001), a 5-year incidence of 1.21% vs 0.18% (p < 0.001) and a 10-year incidence of 4.18% vs 0.36% (p < 0.001). Liver transplant recipients exhibited an elevated risk for the development of NMSC, which persisted and increased over time (incidence of 0.09% vs 0.04% at 1 year (p < 0.001), 0.83% vs 0.18% at 5 years (p < 0.001) and 2.65% vs 0.36% at 10 years (p < 0.001)). Heart transplant recipients also showed a significantly higher cumulative incidence of NMSC at 1 year (0.09% vs 0.04%, p < 0.001), 5 years (0.89% vs 0.18%, p < 0.001) and 10 years (4.06% vs. 0.36%, p < 0.001) post-transplantation. Organ transplant recipients have an 2 times at 1 year, 4,5 times after 5 years and 9 times after 10 years increased risk of NMSC on average as opposed to general Polish population in the same period.

Performance of ultra-high-frequency ultrasound in the evaluation of skin atrophy in patients with long-term oral glucocorticoid therapy in a tertiary rheumatology center.

Ultra-high-frequency ultrasound (UHF-US) allows visualization of the epidermis, dermis, and subcutis and precise measurement of skin thickness. The aim of this study was to assess the performance and interobserver reliability of UHF-US for measuring skin thickness in patients with long-term systemic glucocorticoid (GC) therapy compared to patients without GC therapy or treated for a shorter period.156 patients with known or suspected inflammatory rheumatic diseases underwent US evaluation for skin thickness by 3 experts in 3 anatomical sites (hand, distal, and proximal forearm). 87 patients were classified as "frequent users" who had received continuous oral GCs for at least one year or at least 3 years with various interruptions. 69 patients without any oral GC therapy in the past or treated for a shorter period were classified as "non-frequent users".UHF-US allowed a precise measurement of skin thickness. Skin thickness at all 3 anatomical sites was significantly decreased in "frequent users" of GCs compared to "non-frequent users" (distal and proximal forearm: p < 0.001; hand: p < 0.05). At all 3 anatomical sites, skin thickness was decreased in patients with clinically assessed parchment-like skin compared to patients without parchment-like skin (distal and proximal forearm: p < 0.001; hand: p < 0.05). Interobserver variability was excellent [hand intraclass correlation coefficient (ICC) = 0.99; proximal forearm ICC = 0.85; distal forearm ICC = 0.84].These data support the idea of UHF-US as an objective and reliable imaging tool for monitoring skin atrophy as adverse effects of GC therapy.

Astaxanthin Prevents Glucocorticoid-Induced Femoral Head Osteonecrosis by Targeting Ferroptosis through the JAK2/STAT3 Signaling Pathway.

Glucocorticoid (GC) is extensively used in clinical practice, and the osteonecrosis of the femoral head caused by them is a common issue in orthopedic surgery, yet the underlying mechanisms remain unclear. Astaxanthin (AST), a potent natural antioxidant, has an unexplored impact on GC-induced osteonecrosis of the femoral head (GIONFH). This study explores the effects and mechanisms of AST in counteracting dexamethasone (Dex)-induced ferroptosis and GIONFH. We developed a rat model of GIONFH using intraperitoneal Dex injections and conducted in vitro analysis by culturing osteoblasts (OBs) with Dex treatment. We assessed the impact of AST on Dex-treated OBs using C11-BODIPY and FerroOrange staining, mitochondrial functionality tests, and protein expression analyses through Western blot and immunofluorescence. The influence of AST on bone microarchitecture of femoral head in rat was assessed using micro-CT, hematoxylin and eosin staining, immunofluorescence, and immunohistochemistry at imaging and histological levels. Our findings suggest that AST exerts an inhibitory effect on Dex-induced ferroptosis and GIONFH. In vitro, AST treatment increased glutathione and decreased malondialdehyde, lipid peroxidation, and mitochondrial-reactive oxygen species. Additionally, AST treatment also enhances the phosphorylation of STAT3, upregulates glutathione peroxidase 4 and osteogenic-related proteins, and stimulates bone formation. To delve deeper into the mechanism, the findings revealed that AST triggered activation of JAK2/STAT3 signaling. Moreover, the use of siRNA-STAT3 blocked the beneficial effect of AST in OBs cultivated with Dex. In brief, AST combats GIONFH by activating the JAK2/STAT3 pathway to inhibit ferroptosis.

Short-Term Metformin Protects Against Glucocorticoid-Induced Toxicity in Healthy Subjects: A Randomized, Double-Blind, Placebo-Controlled Trial.

Glucocorticoids (GCs) are potent anti-inflammatory drugs, but strategies to prevent side effects are lacking. We investigated whether metformin could prevent GC-related toxicity and explored the underlying mechanisms. This single-center, randomized, placebo-controlled, double-blind, crossover trial compared metformin with placebo during high-dose GC treatment in 18 lean, healthy, male study participants. The trial was conducted at the University Hospital Basel, Switzerland. Participants received prednisone 30 mg/d in combination with metformin or placebo for two 7-day periods (1:1 randomization). The primary outcome, change in insulin sensitivity, was assessed using a two-sided paired t test. Before and after each study period, we conducted a mixed-meal tolerance test, blood metabolomics, and RNA sequencing of subcutaneous adipose tissue biopsy specimens. Metformin improved insulin sensitivity as assessed by the Matsuda index (n = 17; mean change: -2.73 ± 3.55 SD for placebo, 2.21 ± 3.95 for metformin; mean difference of change -4.94 [95% CI, -7.24, -2.65)]; P < 0.001). Metabolomic and transcriptomic analyses revealed that metformin altered fatty acid flux in the blood and downregulated genes involved in fatty acid synthesis in adipose tissue. Metformin reduced markers of protein breakdown and bone resorption. Furthermore, metformin downregulated genes responsible for AMPK inhibition and affected GLP1 and bile acid metabolism. Metformin prevents GC-induced insulin resistance and reduces markers of dyslipidemia, myopathy, and, possibly, bone resorption through AMPK-dependent and -independent pathways.

Activation of Local 11β-Hydroxysteroid Dehydrogenase Type 1 by Diosmetin Enhances Endogenous Glucocorticoid Levels to Alleviate Skin Inflammation: Insights Into a Novel Therapeutic Strategy for Atopic Dermatitis.

Glucocorticoids (GCs) are synthesised de novo by peripheral tissues and the adrenal cortex of the hypothalamic-pituitary-adrenal axis. Skin expresses an enzyme called 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which reduces cortisone to the active hormone cortisol which activates GC receptors. 11β-HSD1 plays a significant role in alleviating atopic inflammation through the elevation of the concentrations of active GC in the skin. This study aimed to investigate the role of diosmetin as an activator of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). In human keratinocytes, diosmetin was found to upregulate 11β-HSD1 protein expression and cortisol levels, as well as the transcriptional expression of 11β-HSD1 mRNA. However, this upregulation of 11β-HSD1 mRNA was abrogated in keratinocytes transfected with 11β-HSD1 small interfering RNA (siRNA). In an atopic dermatitis (AD) murine model, topical administration of diosmetin significantly attenuated basal transepidermal water loss (TEWL) and the Eczema Area and Severity Index (EASI), while enhancing stratum corneum (SC) hydration. Diosmetin also increased corticosterone levels in the SC and upregulated 11β-HSD1 expression in both the serum and epidermis. Furthermore, diosmetin treatment led to a marked reduction in serum immunoglobulin E (IgE) and tumour necrosis factor-α (TNF-α) levels, and suppressed mRNA expression of thymic stromal lymphopoietin (TSLP), interleukin-1β (IL-1β), IL-4, and IL-13 in the epidermis. Collectively, these findings suggest that diosmetin promotes the endogenous activation of glucocorticoids via local 11β-HSD1 activation, underscoring its potential as a novel topical therapeutic agent for the management of inflammatory skin disorders, such as AD.

New anti-inflammatory mechanism of glucocorticoids uncovered

Glucocorticoids (GCs) are potent anti-inflammatory drugs. A new study by Auger et al. found that GCs increase itaconate, an anti-inflammatory tricarboxylic acid (TCA) cycle intermediate, by promoting movement of cytosolic pyruvate dehydrogenase (PDH) to mitochondria. Itaconate was sufficient for mediating the anti-inflammatory effects of GCs in mice, overriding the notion that nuclear glucocorticoid receptor (GR) is necessary for inflammation inhibition.

Network Pharmacology and Experimental Validation of Jinwei Decoction for Enhancement of Glucocorticoid Anti-Inflammatory Effect in COPD through miR-155-5p.

Jinwei decoction can enhance the anti-inflammatory effect of glucocorticoid (GC) on chronic obstructive pulmonary disease (COPD) by restoring the activity of human histone deacetylase-2 (HDAC2). However the upstream mechanism of Jinwei decoction on HDAC2 expression is not clear. To explore the target of Jinwei decoction to enhance the anti-inflammatory effect of GC on COPD through microRNA155-5p (miR-155-5p) by network pharmacology and experimental verification. The TCMSP database was used to screen active ingredients and target genes of Jinwei decoction, and miRWalk2.0 was used to predict downstream target genes of miR-155-5p. COPD-related genes were identified by searching GeneCards, Grugbank and OMIM databases; Venny 2.1 was used to screen intersection genes; Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of intersection genes were analyzed by R software. Protein-protein interactions (PPIs) were analyzed by Cytoscape 3.7.2 software to identify core genes. Finally, interactions between main compounds and potential targets were verified by molecular docking. A COPD cell model was established by 5% cigarette smoke extract (CSE)- induced bronchial epithelial cell (BEAS-2B), and the results of network pharmacology were verified by in vitro experiments. Two hundred thirty-one active ingredients, 352 Jinwei decoction drug targets, 5949 miR-155-5p target genes, 8286 COPD target genes, and 127 intersection genes were identified. Twelve core proteins of PPI networks may be involved. GO enrichment analysis showed that regulation of membrane potential， response to steroid hormone, and histone modification were involved; KEGG pathway enrichment analysis concentrated in the PI3K-Akt, mitogen-activated protein kinase (MAPK), HIF-1, and other signaling pathways. The molecular docking results showed that quercetin, luteolin and stigmasterol have higher affinity with PTGS2, HIF1A and AKT1. The results of cell experiments revealed that Jinwei decoction not only enhances the anti- inflammatory effect of GC in the COPD cell model but also reverses the high expression of miR-155-5p、PI3k、Akt, and low expression of HDAC2, thereby inhibiting the inflammatory response of COPD. Jinwei decoction can regulate HDAC2 activity and enhance the anti-inflammatory effect of GC on COPD by modulating miR-155-5p. Its mechanism of action may be related to its effect on the PI3K-Akt through miR-155-5p.

Two Decades Rituximab Therapy in Anti-Neutrophil Cytoplasmic Antibody Associated Vasculitis.

Remission failure and relapse numerate as one of the main problems in patients with antineutrophil cytoplasmic antibody-associated vasculitis (AAVs). The need for new agents that provide effective and safe induction accompanied by sustained remission seems to be urgent in clinical care. The efficacy and safety of rituximab (RTX) for AAVs therapy has been reported in various studies. RTX therapy offers several advantages to treating AAVs patients compared to other therapeutic approaches including reduction of Glucocorticoids (GCs) and conventional Immunosuppressive therapy (IST) usage during both the induction of remission and maintenance phases. This reduction can lead to a lower rate of serious complications making RTX therapy a safer option. It seems that RTX may provide improved clinical outcomes in these patients mediated via B-lymphocyte depletion, Proteinase 3-antineutrophilic cytoplasmic antibody (PR3-ANCA), and myeloperoxidase-antineutrophilic cytoplasmic antibody (MPO-ANCA) titers reduction. In this regard, some uncertainties have been reported to validate the association between such depletion and clinical improvement, as suggested by other sources of autoreactive B cells that did not target with RTX. Due to the prolonged B cell depletion, fixed intervals and adjusted dosage of RTX may be required in patients with AAVs. In this narrative review, we aimed to insight better understand regarding the efficacy of RTX for effective induction and sustained remission in patients with AAVs. It seems that discovering new biomarkers predicting relapse in AAVs patients can lead to future targeted therapy.

C16 peptide and angiopoietin-1 alleviate the side effects of glucocorticoids in a rat multiple sclerosis model.

Natural glucocorticoids (GCs) have been widely used to treat acute multiple sclerosis (MS) attacks. However, they also cause significant side effects related to immunosuppression. Our previous study found that C16 peptide combined with angiopoietin-1 (Ang-1) inhibited inflammatory cell infiltration and protected blood vessels in animal models of inflammatory neurodegenerative diseases. An acute experimental autoimmune encephalomyelitis model was established in Lewis rats to explore the effects of these drugs on MS. One hundred rats were equally and randomly assigned into five groups: normal control, vehicle, low-dose methylprednisolone (MP), high-dose MP, and C16+Ang-1 (C+A). Histological examinations, behavioral tests, and high-throughput 16S rRNA gene sequencing were conducted to determine inflammation levels in the central nervous system, neuronal survival, functional recovery and gut microbiota. The results illustrated that C+A exerted a neuroprotective effect in MS rats, with fewer side effects observed in the C+A group than in the high-dose MP group. The abundance of Campylobacter was increased in vehicle-treated rats, indicating an imbalance of the gut microbiota after MS. The abundance of probiotic Lactobacillus plantarum was increased in the C+A group. Low-dose MP failed to reverse the gut microbiota imbalance, whereas both the C+A and high-dose MP groups exhibited gut microbiota profiles more similar to those of the normal controls, with C+A displaying superior efficacy. C16 plus Ang-1 might serve as a complement to GCs for the treatment of MS. Changes in the abundance of Campylobacter and L. plantarum suggest their essential roles in the pathogenesis of MS.

Exploration of rituximab treatment strategies for membranous nephropathy adapted to the Chinese healthcare environment

PurposeThis study aimed to explore the specific efficacy of rituximab (RTX) in the treatment of membranous nephropathy (MN) and compare and analyze the differences in effectiveness among various treatment regimens, with the objective of identifying the optimal treatment protocol suitable for the medical environment in China.Patients and methodsThis retrospective study focused on patients with MN who were treated with RTX and hospitalized at the First Medical Center of PLA General Hospital between January 1, 2019, and December 30, 2022. These patients were followed up for more than one year. We collected clinical data from these patients and categorized them into three groups on the basis of their RTX treatment background: the combined glucocorticoids (GCs) and/or immunosuppressants (IMS) and RTX monotherapy treatment groups, the initial and non-initial treatment groups, and the standard RTX and non-standard RTX treatment groups. The study evaluated the comprehensive outcomes of complete or partial remission during follow-up, as well as relapses after remission. Additionally, Cox regression analysis was conducted to identify risk factors influencing patient remission and relapse.ResultsA total of 126 patients were enrolled in this study, with an average age of 49.0 ± 13.4 years. Among them, males accounted for up to 77.8%, with an average BMI of 26.7 ± 4.0. Among these patients, 59.5% (75/126) received RTX combined with GCs and/or IMS. Statistical results revealed that the combined use of GCs and/or the IMS had no significant effect on renal remission (P = 0.439), but it accelerated the process of renal remission (P = 0.010). A total of 34.9% (42/126) of patients chose RTX as the initial treatment. Compared with the non-initial treatment group, this choice did not significantly differ in terms of efficacy or faster remission speed (all P > 0.05). On the other hand, 39.7% (50/126) of patients received the standard RTX treatment regimen. Compared with the non-standard group, the standard RTX treatment group presented a better remission rate (P < 0.001) and a faster remission speed (P = 0.027). During 13.0 (12.0, 20.0) months of follow-up, the cumulative remission rate reached 73% (92/126), including 47.6% (60/126) of patients with partial remission (PR) and 25.4% (32/126) of patients with complete remission (CR). The cumulative relapse rate was 20.7% (26/126). In addition, 17.5% (22/126) of patients experienced adverse reactions. Multivariate Cox regression analysis revealed that the standard RTX treatment regimen was associated with a better remission rate, whereas comorbid diabetes reduced the remission rate. Older age and higher white blood cell counts may lead to a higher relapse rate.ConclusionThis study revealed that RTX treatment has a high remission rate and a low relapse rate in MN patients. The standard RTX treatment regimen can provide better benefits. However, our experience is limited by its retrospective design and relatively small sample size, and further large-scale randomized controlled studies are needed to confirm our preliminary findings.

Review of Current First-Line Pharmacotherapy Strategies for Thyroid Eye Disease

Thyroid eye disease (TED) is a challenging autoimmune disorder that often requires therapeutic intervention. Glucocorticoids (GCs), particularly in intravenous (IV) regimens, remain the first-line treatment due to their strong anti-inflammatory and immunosuppressive effects. This paper focuses on the efficacy, protocols, and safety profiles of selenium supplementation and glucocorticoid therapy, emphasizing its central role in TED management. Alternative therapies, such as teprotumumab, rituximab, and tocilizumab, are briefly discussed as potential options for patients with moderate-to-severe TED, who are unresponsive or intolerant to glucocorticoids. Adherence to recommended dosing protocols and careful monitoring of adverse effects are essential to optimize treatment outcomes.

Response to initial treatment with glucocorticoids in TAFRO syndrome and implications for secondary treatment.

The study aimed to investigate the therapeutic effect of various initial treatments incorporating glucocorticoid (GC) in TAFRO syndrome (thrombocytopenia, anasarca, fever, reticulin fibrosis, and organomegaly). Cases of TAFRO syndrome up to November 2023 were retrospectively collected. Overall survival (OS) and resistance to GC therapy were assessed, with resistance analyzed based on the time to the next treatment or death (TTNTD). The study included 95 patients, including 5 diagnosed at our hospital. OS did not differ significantly between patients who received GC monotherapy and those who had a second-line therapy added within 2weeks (100-day OS rate: 86.6% vs. 77.7%; p = 0.338). Moreover, 100-day OS did not differ between patients who received GC pulse therapy within 2weeks and those who did not (77.5% vs. 93.1%, p = 0.129). In multivariate analyses, pretreatment severity score ≥ 8 (hazard ratio [HR], 2.99; 95% confidence interval [CI] 1.05-8.50) and platelets ≥ 6.9 × 10^4/µL (HR, 2.26; 95% CI 1.01-5.02) were significantly associated with shorter TTNTD. Additional second-line or GC pulse therapy provided no advantage in the hyperacute phase. Higher severity scores and platelet values may predict resistance to GC therapy.

P-2294. Listeriosis in cancer patients: Uncommon and protean

Abstract Background Historically cancer patients (pts) are considered to be at an increased risk of Listeria monocytogenes infection (listeriosis, LT), especially those with ongoing chemotherapy or receipt of glucocorticoids (GCs). However, the features of this uncommon infection have not been described in contemporary cohorts of cancer pts. Methods We performed a 12-year retrospective chart review of cancer pts with LT as confirmed by positive culture from blood or other sterile clinical samples at MD Anderson Cancer Center (2011-2023). Results We identified 20 cancer pts with LT. Twelve (60%) had an underlying hematologic malignancy (Figure 1). Lymphopenia &amp;lt; 1000/µL (17, 85%, severe &amp;lt; 200/µL in 20%) and monocytopenia &amp;lt; 1000/µL (18, 90%, severe &amp;lt; 200/µL in 35%) were common at diagnosis, as well as chemotherapy (13, 65%) and GC use (11, 55%) within 30 days prior to LT diagnosis. Most (15 pts) were &amp;lt; 65 years old. Fever was seen in 10 pts. Vomiting/diarrhea were common (50%), while 45% of pts had headache or altered mental status. LT most often presented with bacteremia (17, 85%) and gastroenteritis (9, 45%) (Figure 2). 7/9 pts with gastroenteritis were bacteremic. Central nervous system (CNS) manifestations (meningitis, brain abscess) were documented in 2 pts who had a lumbar puncture. Focal non-CNS infections were seen in 30% of pts in unusual sites: cellulitis, abscess, endocarditis, cholangitis, urinary tract infection, and peritonitis. Of interest, 11pts (55%) had low SOFA score at presentation and were given antibiotics only after blood cultures were positive for Listeria. In fact, most pts (17, 85%) either did not receive empiric (n=11) or had inappropriate empiric antibiotics (n=6) until LT diagnosis. The 30-day mortality from LT diagnosis was 25% and correlated with high SOFA score ≥ 5 (5/8 pts, P = 0.0005). Delayed appropriate antibiotics were not significantly associated with increased 30-day mortality in this small cohort (Table 1). Conclusion Although uncommon, LT in cancer pts has pleotropic manifestations and variable morbidity, from non-bacteremic gastroenteritis to septicemia, CNS and non-CNS focal infections. Consider LT in ill cancer pts with lymphopenia/monocytopenia and recent chemotherapy or GCs, especially if they have GI symptoms, even in the absence of fever or CNS involvement. Disclosures Sebastian Wurster, MD, MSc, Astellas Pharma: Grant/Research Support|Gilead Sciences: Grant/Research Support Dimitrios P. Kontoyiannis, MD, AbbVie: Advisor/Consultant|Astellas Pharma: Advisor/Consultant|Astellas Pharma: Grant/Research Support|Astellas Pharma: Honoraria|Cidara Therapeutics: Advisor/Consultant|Gilead Sciences: Advisor/Consultant|Gilead Sciences: Grant/Research Support|Gilead Sciences: Honoraria|Knight: Advisor/Consultant|Merck: Advisor/Consultant|Scynexis: Advisor/Consultant

Using Faecal Cortisol Metabolites to Assess Adrenocortical Activity in Wild-Living Alpine Marmot Marmota marmota: A Biological Validation Experiment.

Faecal cortisol/corticosterone metabolites (FCMs) have become increasingly popular as an easy-to-sample, non-invasive and feedback-free alternative to assess glucocorticoid (GC) levels, key components of the neuroendocrine stress response and other physiological processes. While FCMs can be a powerful aid, for instance, for gaining insights into ecological and evolutionary processes, as well as to assess animal welfare or impacts of anthropogenic stressors on wildlife populations, this method comes with specific challenges. Because GCs are heavily metabolised before excretion, it is critical to validate the enzyme immunoassays (EIAs) used to measure FCMs. Additionally, because species may differ in metabolite profiles, assay validation must be performed separately for each focal species. Despite this, the use of unvalidated assays remains widespread. We performed a biological validation experiment to test a set of EIAs to measure FCMs and adrenocortical activity in free-living Alpine marmots Marmot marmota. We capitalised on capture and handling as part of a relocation project of marmots under the assumption that capture, and handling represent a stressful event and tracked changes in FCM levels over the following 48 h. Faeces samples collected at capture were assumed to return baseline FCM levels. Of the three EIAs tested, only the 11-oxoetiocholanolone '72T' EIA detected an increase in FCM levels about 18 h after capture. This result paves the way for future studies using FCMs to investigate the adrenocortical activity in this species.

Isovitexin targets SIRT3 to prevent steroid-induced osteonecrosis of the femoral head by modulating mitophagy-mediated ferroptosis

The death of osteoblasts induced by glucocorticoid (GC)-mediated oxidative stress plays a crucial role in the development of steroid-induced osteonecrosis of the femoral head (SIONFH). Improving bone formation driven by osteoblasts has shown promising outcomes in the prognosis of SIONFH. Isovitexin has demonstrated antioxidant properties, but its therapeutic effects on GC-induced oxidative stress and SIONFH remain unexplored. In this study, we analyzed clinical samples obtained from SIONFH patients using proteomic and bioinformatic approaches. We found an imbalance in mitochondrial homeostasis and ferroptosis-induced impairment of osteogenic capacity in SIONFH. Subsequently, we investigated the cause-and-effect relationship between mitochondria and ferroptosis, as well as the regulatory role of mitophagy in maintaining mitochondrial homeostasis and controlling ferroptosis. We then identified the critical involvement of SIRT3 in modulating mitochondrial homeostasis and ferroptosis. Furthermore, molecular docking and co-immunoprecipitation confirmed the strong interaction between SIRT3 and BNIP3. Strikingly, restoring SIRT3 expression significantly inhibited pathological mitophagy mediated by the BNIP3/NIX pathway. Additionally, we discovered that Isovitexin, by promoting SIRT3 expression, effectively regulated mitophagy, preserved mitochondrial homeostasis in osteoblasts, suppressed ferroptosis, and restored osteogenic capacity, leading to remarkable improvements in SIONFH. These findings reveal the effects and molecular mechanisms of Isovitexin on SIONFH and highlight the potential of targeting SIRT3 as a promising strategy to suppress mitophagy-mediated ferroptosis in osteoblasts and against SIONFH.

Glucocorticoids on bone remodeling in systemic lupus erythematosus mice.

Systemic lupus erythematosus requires glucocorticoids for management. This study investigates how glucocorticoids influence bone in a SLE mouse model, focusing on bone mineral density (BMD), microstructure, and remodeling markers. MRL/lpr and C57BL/6 mice were administered dexamethasone or saline as a control for 4-weeks. Bone assessments included analyses of BMD, bone structure, and serum levels of RANKL and OPG. Dexamethasone decreased BMD and altered cortical and trabecular bone thickness in both MRL/lpr and C57BL/6 mice. In C57BL/6 mice, cortical bone exhibited increased catabolism while trabecular bone showed signs of increased anabolism, whereas MRL/lpr mice did not show significant changes in bone turnover. Both strains experienced weight loss, with a significant decrease in femur length observed only in C57BL/6 mice. Dexamethasone exacerbated BMD reduction in MRL/lpr mice and halted its increase in C57BL/6 mice. C57BL/6 mice exhibited notable changes in cortical and trabecular bone structure, while MRL/lpr mice didn't. After receiving dexamethasone, both strains showed higher serum RANKL levels, especially in C57BL/6 mice. OPG decreased in both strains. Both glucocorticoids and SLE contribute to abnormal bone remodeling through RANKL/OPG pathway. Glucocorticoid (GC) treatment in a mouse model of systemic lupus erythematosus (SLE) leads to significant changes in bone parameters, including decreased bone mineral density (BMD) and alterations in bone structure. Those change are associated with the modulation of RANKL and OPG expression. Both GC and inflammation in SLE contribute to BMD reduction, and GC may have a certain protective effect on bone in the early stage of chronic inflammation. GC can upregulate RANKL expression and downregulate OPG expression in vivo. During a state of chronic inflammation, RANKL expression increases. However, OPG may not exert a significant influence on inflammatory stimulation.

Impact of Repeated Glucocorticoid Oral Administration on the Urinary Steroid Profile.

The detection of endogenous anabolic androgenic steroids (EAAS) is performed with the Steroidal Module of the Athlete Biological Passport (ABP). Glucocorticoids (GC) could be a confounding factor to the ABP Steroidal Module because they inhibit the hypothalamic-pituitary-adrenal axis, and ABP metabolites have partial adrenal origin. In previous studies, single-dose systemic GC administrations have been shown to reduce the urinary ratios A/T and 5αdiol/E. In this work, the impact of repeated oral doses of GCs on the urinary steroid profile (SP) has been evaluated. The treatments administered consisted of multiple oral administrations of methylprednisolone (12 mg/24 h for 3 days, n = 8) and dexamethasone (2 mg/12 h for 5 days, n = 8). Urine samples were collected before, during, and after the GC treatments, and the SP was measured in all samples using gas chromatography-tandem mass spectrometry. The multiple-dose oral administration of GCs resulted in a treatment-dependent reduction of the excretion rates of some urinary metabolites (5αAdiol, A, and Etio) and the urinary SP ratios A/T and 5αAdiol/E. The T/E ratio was not significantly affected. Overall, although the consumption of GC could result in atypical profiles for A/T and 5αAdiol/E ratios, according to the cost/benefit assessment, GC should not be considered a confounding factor to the urinary SP because misunderstandings would only take place in very specific situations and, in those cases, the analysis by isotope ratio mass spectrometry of the urine triggering the atypical profile would demonstrate the endogenous origin of the SP metabolites.

P0513 Comparative Evaluation of Clinical Scores for Predicting Response to Systemic Steroid Treatment in Hospitalized Adult Patients with Acute Severe Ulcerative Colitis: A Retrospective Study

Abstract Background Approximately 30% of hospitalized patients with acute severe ulcerative colitis (ASUC) may be refractory to systemic glucocorticoid (GC) therapy, administered orally or intravenously necessitating salvage therapy 1. Clinical tools predicting response to GC could allow for early initiation of salvage therapy thus reducing GC exposure and improving overall outcomes. However, comparative studies for available prognostic scores are limited and yield conflicting results 2,3. We aimed to determine the best performing score on a cohort of patients with ASUC from a large tertiary hospital. Methods A retrospective cohort study was conducted between 2017-2022. Hospitalized patients with ASUC, as defined by Truelove and Witts criteria and treated with GC, were included. Patients were classified as GC-refractory (GC-Ref) if they had no clinical and biochemical response, requiring salvage therapy. Assessments for GC response were performed on the third and seventh day of hospitalization. The predictive value of the Mayo endoscopic and Adams scores was evaluated on admission, while the Gibson, Ho, Lindgren, and Oxford scores were assessed on the third day. We compared scores using a multivariate logistic regression and ROC analysis. Results Ninety-six patients met the inclusion criteria and were included, 56 (58%) were GC-responsive (GC-Res) and 40 (42%) were GC-Ref. The mean age of admission was similar between the groups (GC-Res: 42.2 ± 16.7 years; GC-Ref: 37.15 ± 15.3; P=0.1). The GC-Ref group had more extensive disease (65% Vs 38.2%, P=0.01), (Table 1). On admission, the GC-Ref group had significantly more bowel movements per day (14.6 Vs 11.3, P=0.05), elevated CRP levels (91.5 mg/L Vs 58.4 mg/L, P=0.005), higher MAYO score (2.8 Vs 2.3, P=0.028) and lower albumin levels (32 g/L Vs 35.4 g/L, P=0.008) and body mass index (20.3 kg/m2 Vs 23.8 kg/m2, P=0.02). The Gibson score (based on three or more stools per day and a CRP-to-albumin ratio (CAR) above 0.85) and Mayo Endoscopic Score demonstrated appropriate sensitivity and significant predictive value for identifying refractory patients (aOR: 11.3 (95% CI 1.87-68), P=0.008 and aOR: 7.9 (95%CI 1.8-34), P=0.006); respectively. The Gibson score, showed the best overall performance with a sensitivity of 0.7, specificity of 0.75 and positive predictive value of 0.71 (Table 2). Of the individual elements of the various scores, CAR measured on the third day of hospitalization had the highest predictive value (aOR: 12.9 (95% CI 2.5-67), P=0.002) with sensitivity of 0.72, and specificity of 0.65. Conclusion In our cohort, the Gibson score proved to be the most effective predictive tool for identifying patients with steroid refractory ASUC. This need to be confirmed in larger prospective cohorts.

P0137 Primary bile acids promote epithelial necroptosis via Stat1 signaling

Abstract Background Gut barrier dysfunction and microbial dysbiosis play a crucial role in the pathogenesis of Inflammatory Bowel Disease (IBD), including Ulcerative colitis (UC) and Crohn´s disease (CD)1. Although IBD is marked by dynamic changes in the intestinal microbiome, little is known about the corresponding changes in the gut metabolism, the molecular interface between the host and its microbiota. In CD, changes in bile acid (BA) metabolism and composition have been observed, with a notable the enrichment of the primary BA chenodeoxycholic acid (CDCA) specifically in CD2, suggesting a potential involvement in disease pathogenesis. However, it is still poorly understood how BAs influence epithelial homeostasis. Methods Serum bile acid (BA) profiles of CD patients and healthy controls were analyzed using BA-targeted metabolomics. Data normalization and statistical analyses were performed to identify significant differences in BA composition between groups. Human ileal organoids were established from biopsy samples obtained from CD patients, while murine small intestinal organoids were derived from wildtype, Stat1-/-, and farnesoid X receptor (FXR) deficient mice. Bulk RNA sequencing was performed on organoids treated with CDCA to investigate transcriptional changes and identify molecular mechanisms underlying CDCA-induced toxicity. A cell death inhibitor screen was applied to delineate the pathways contributing to CDCA-induced IEC toxicity. Results Our findings reveal a significant difference in BA profiles between CD patients and healthy controls, with a notable enrichment of the primary BA, CDCA, in CD patients. CDCA exerts a pronounced toxic effect on both human and murine small intestinal organoids (SIO) in a dose-dependent manner, whereas the secondary BA, deoxycholic acid (DCA), does not. Importantly, the cytotoxicity of CDCA was markedly reduced when it was conjugated with glycine or taurine, suggesting that conjugation mitigates CDCA's impact on IEC viability. Mechanistically, CDCA disrupts BA signaling pathways, enhances the interferon (IFN) response, and activates cell death pathways within SIO models. CDCA specifically promotes epithelial necroptosis via STAT1 signaling, highlighting a pathway through which CDCA may exacerbate epithelial damage in CD. Therapeutically, both tofacitinib (Tofa) and glucocorticoids (GCs) demonstrate a protective effect, significantly suppressing the CDCA-induced IFN response. These findings suggest that modulation of BA signaling and inhibition of the IFN response may be promising strategies to protect intestinal epithelial integrity in CD patients. Conclusion These findings suggest that modulation of BA signaling may be promising strategies to protect intestinal epithelial integrity in CD patients.