Carbohydrates with high structural complexity are involved in key biological processes and serve as susceptible targets of environmental metabolic disruptors. However, the lack of a high-throughput and sensitive method for profiling carbohydrates limits our mechanistic understanding of the disruption of sugar metabolic pathways. In this study, we found that postcolumn addition of 10 μM (chloromethyl)triphenylphosphonium chloride (Ph3(CH2Cl)PCl) enabled the efficient ionization of saccharides and sugar alcohols as [M+Cl]- adducts in high-resolution mass spectrometry analysis, lowering instrumental detection limits by an average of 4- to 767-fold. A nontargeted method for polyol-containing compounds (PCCs) was subsequently established by leveraging their characteristic chlorinated isotope patterns. Approximately 68 PCCs, including saccharides with a degree of polymerization of 1-15, sugar alcohols, nonglucosyl saccharides, O-glycosides, nucleosides, amino sugars, a purine base, and a vitamin, were identified from microbially rich samples such as beer and mouse cecal contents. The method successfully identified disrupted saccharides and nucleosides in the cecal contents of mice exposed to a bisphenol A replacement (TGSA). Ph3(CH2Cl)PCl-enhanced ionization was further found to enhance analytical sensitivity and expand the species coverage in mass spectrum imaging, broadening the range of detectable saccharides (e.g., di- and trisaccharides) in biological samples. The imaging method confirmed alterations of PCCs in the central nervous system and liver tissues by medium-chain chlorinated paraffins during fetal development. This study provides an effective tool for the quantification, identification, and imaging of PCCs to screen for environmental disruptors of sugar metabolism.

BackgroundBody mass index (BMI) has been associated with improved outcomes in several malignancies, including prostate cancer. However, the impact of BMI on treatment efficacy, particularly with androgen receptor pathway inhibitors (ARPIs), in metastatic hormone-sensitive prostate cancer (mHSPC) remains insufficiently characterized. This study aimed to evaluate the association between BMI and radiographic progression-free survival (rPFS) in patients with mHSPC receiving first-line therapies.MethodsWe retrospectively analyzed 330 patients with newly diagnosed mHSPC. Patients were stratified by their treatment regime and baseline BMI. rPFS was assessed using Kaplan-Meier analysis and Cox proportional hazards models. Multivariate analyses included clinically relevant covariates and factors significant in univariate analyses.ResultsWith a median follow-up of 47 months, median rPFS for the entire cohort was 34 months. Patients with BMI≥25 kg/m[2] demonstrated significantly longer median rPFS. Higher BMI was associated with favorable baseline characteristics, including better ECOG performance status and higher hemoglobin levels. In multivariate analysis, liver metastases, high-volume disease, low hemoglobin, elevated alkaline phosphatase, and treatment modality remained independent predictors of rPFS, whereas BMI did not retain independent significance. Among overweight patients receiving ARPIs, enzalutamide showed a numerically longer rPFS compared with abiraterone, although this difference was not statistically significant.ConclusionHigher BMI was associated with prolonged rPFS in patients with mHSPC, supporting a potential obesity paradox in this population. While BMI was not an independent predictor in multivariate analysis, it may reflect underlying patient fitness and disease biology. These findings warrant prospective validation into metabolic factors influencing ARPI efficacy.

Liver-specific ERα knockout (LERKO) mice with normal ovarian function are not susceptible to steatosis and maintain mitochondrial function. Here, we tested whether loss of hepatic ERα blunts estrogen modulation of hepatic mitochondrial respiratory capacity and mitochondrial proteome following ovariectomy (OVX). Sham or OVX surgery was performed in middle-aged female mice (36-40 weeks), followed by AAV injection to generate Control (Con; GFP) or LERKO mice (Cre). All mice were placed on a high-fat diet (HFD) for 10 weeks following surgery. Half of the OVX mice received 17-beta estradiol (E2) replacement (OVX + E2) for the last 4 weeks. OVX mice had greater body mass and adiposity, which was reversed by E2 replacement in both Con and LERKO mice. E2 replacement reduced steatosis in both Con and LERKO OVX mice, but the LERKO OVX mice maintained greater hepatic triglycerides. E2 replacement promoted greater basal and ADP-stimulated (State 3) mitochondrial respiration in Con OVX but not in LERKO OVX mice. Changes in mitochondrial respiration could not be attributed to altered responses to changes in energy demand (GATP) or to alterations in mitochondrial H2O2 production. Conversely, maximal coupled branched-chain amino acid-supported respiration was universally suppressed by E2 replacement. Proteomics analysis revealed E2-mediated reductions in hepatic mitochondrial energy transduction in both Con and LERKO mice. In conclusion, post-ovariectomy estrogen treatment reduces steatosis in the absence of hepatic ERα; however, steatosis remains higher, and mitochondrial deficits persist despite similar proteomic changes, suggesting ERα is required for optimal response to estrogen in modulating hepatic mitochondrial respiration.

Isolated polycystic liver disease (PLD) is rare and may cause debilitating mass-effect symptoms despite relatively preserved hepatic synthetic function. Because the Model for End-Stage Liver Disease (MELD) score does not capture nutritional, functional, and mechanical disease burden, affected patients may have low calculated MELD scores despite advanced symptomatic disease. Updated MELD exception guidance broadened consideration for PLD to include malnutrition and sarcopenia. We describe a patient with isolated PLD who was initially considered too well for transplantation but subsequently developed progressive weight loss, sarcopenia, and functional decline, prompting repeat evaluation, wait-list registration, and MELD exception approval. Several weeks after listing, he sustained traumatic hepatic cyst rupture, resulting in fatal hemoperitoneum and abdominal compartment syndrome. This case highlights the evolving clinical course of symptomatic PLD, the importance of longitudinal reassessment, and the potential severity of rare cyst-related complications.

Obesity is a global, complex, and multifactorial disease, and a major risk factor for diabetes, cardiovascular disease, fatty liver disease, and cancer. Despite its high prevalence, effective diagnostic and therapeutic strategies remain limited. Conventional anthropometric measurements, such as body mass index (BMI) and waist circumference, offer limited accuracy in assessing the impact of excess adiposity and related comorbidities on individual patients. Current anti-obesity pharmacotherapies also show inconsistent efficacy, are often associated with systemic side effects, and poor adherence to necessary lifestyle modifications, which limit their long-term success. Emerging evidence underscores the critical role of non-coding RNAs, including microRNAs, long non-coding RNAs, circular RNAs, and tRNA-derived small RNAs in key metabolic processes such as adipogenesis, insulin signalling, and appetite control. Notably, the profiles of many circulating miRNAs have been reported to differ between patients with obesity compared to healthy lean individuals, suggesting their potential as minimally invasive diagnostic biomarkers. In this mini-review, we discuss the clinical promise of RNA-based approaches for disease risk assessment and as therapeutic targets in obesity. In addition, we discuss key challenges, including biomarker validation and effective RNA delivery systems to the target organs. Multi-omics approaches may help overcome many of these limitations and enable the development of targeted and personalized RNA-based anti-obesity therapies.

Hypoglycemia is a frequent finding after simultaneous pancreas-kidney (SPK) transplantation, but its etiology is often multifactorial and challenging to interpret. We report a rare case of late-onset severe hypoglycemia secondary to a neuroendocrine tumor arising in a pancreatic allograft. A 60-year-old man with type 1 diabetes mellitus underwent SPK transplantation in 2007 with immediate graft function and long-term metabolic stability. Ten years post-transplant, partial pancreatic graft dysfunction and progressive vascular complications were noted. In 2024, 17 years after transplantation, the patient developed recurrent, documented hypoglycemic episodes despite withdrawal of glucose-lowering therapy, with fasting plasma glucose values as low as 40 mg/dL and a marked reduction in HbA1c. Biochemical evaluation suggested endogenous hyperinsulinism, with elevated chromogranin A levels. Imaging studies revealed a large hypervascular mass within the pancreatic graft, with high somatostatin receptor expression on 68Ga-DOTA-TOC PET-CT. Histology from biopsy and subsequent total graft pancreatectomy confirmed a well-differentiated grade 2 neuroendocrine tumor with focal insulin expression, consistent with insulinoma. Surgical resection resulted in complete resolution of hypoglycemia. This case highlights the need to consider rare but clinically significant causes of hypoglycemia in SPK recipients and underscores the importance of a comprehensive, multidisciplinary diagnostic approach integrating continuous glucose monitoring, biochemical markers, and advanced imaging.

Micafungin is recommended as a first-line antifungal agent for the treatment of invasive candidiasis. However, the optimal dosing regimen in critically ill patients remains uncertain due to marked interindividual pharmacokinetic variability. This study aimed to develop a population pharmacokinetic (popPK) model for micafungin and an electronic nomogram to support the selection of the optimal initial dosing regimen. A retrospective, monocentric pharmacokinetic study was conducted in a digestive ICU. PopPK modelling and covariate analysis were performed using Monolix.2024R1. Monte Carlo simulations (MCS) were subsequently used to build the e-nomogram. Overall, 404 micafungin plasma concentrations obtained from 100 patients were analysed, including 40% with cirrhosis and/or a history of liver transplantation. The concentration-time data were adequately described by a linear one-compartment model. The estimated PK parameters (between-subject variability) were 17.3 L (15%) for volume of distribution (Vd) and 0.85 L/h (25%) for clearance (Cl). Total body Weight (TBW) and total bilirubin were identified to influence Vd and/or Cl. External model evaluation showed acceptable predictive performance for both population and individual predictions. MCS suggested that the currently recommended regimen of 100 mg/day may be insufficient to achieve the PK/PD target for all Candida species when clinical breakpoint MIC values are considered. The resulting RShiny application provides clinicians with a practical decision support-toolfor selecting the best loading and maintenance doses according to TBW, BILI and the target pathogen, thereby maximising the probability of PK/PD target attainment.

Deep characterization of intact proteoforms remains an analytical challenge in functional proteomics, particularly for heterogenous multi-site post-translational modifications at distinct amino acid residues. Histones are among the most dynamically and diversely post-translationally modified proteins in eukaryote cells, carrying multiple, co-occurring and reversible modifications that can give rise to isomeric proteoform species. Tandem mass spectrometry with multimodal fragmentation capabilities is a promising approach for deep characterization of intact proteoforms, such as modified histones. We applied the novel timsOmni mass spectrometer, which incorporates the Omnitrap platform enabling multimodal MS[n] workflows using controlled in vitro acetylation of recombinant histones H3.1 and H4 by GCN5, PCAF and p300, followed by analysis of endogenous H4 proteoforms. Complementary MS[2] electron- and collision-based dissociation (ECD, EID, RCID and ECciD), together with MS[3] strategies, produced complete or near-complete backbone fragmentation of intact protein ions (>92% amino acid sequence coverage). For monoacetylated species generated by the more site-selective lysine acetyltransferases, the dominant proteoform matched the known catalytic preferences of the enzymes (H3.1K14ac for GCN5 and PCAF, and H4K8ac for PCAF), while minor positional isomers were also identified and their relative abundance estimated. In contrast, the broader substrate specificity of p300 produced a wide distribution of H4 proteoforms bearing up to seven acetylated lysine residues. Species carrying six and seven acetylations were characterized by multimodal MS[2]/MS[3] experiments, enabling localization of individual acetylation sites and discrimination of positional isomers. Finally, top-down sequencing of endogenous intact H4 proteoforms from human liver extracts yielded amino acid sequence coverages of 92-93% for the most abundant species and confident localization of multiple, distant PTMs (acetylation and methylation). These results demonstrate that multimodal MS[n] fragmentation of intact proteins supports residue-level assignment of combinatorial histone marks and coexisting positional isomers in controlled and endogenous proteoform mixtures.

Mancozeb is widely used as a broad-spectrum fungicide, and its potential effects on the gut-liver-kidney axis remain incompletely understood. In this study, we used an experimental oral exposure model to identify dose-associated effects and underlying mechanisms associated with mancozeb exposure. Following exposure to mancozeb at doses of 1, 10, and 100 mg/kg body weight, the fungicide was found to accumulate primarily in the digestive system (including the colon, cecum, and their contents) and feces of mice. Mancozeb exposure may exert adverse effects on the biomarkers related to liver function in mice, as evidenced by elevated activities of alanine aminotransferase and aspartate aminotransferase, increased malondialdehyde levels, and reduced activities of superoxide dismutase and glutathione peroxidase. Additionally, it caused significant shortening of the small intestine and increased intestinal permeability, reflected by elevated wet weight and dry-to-wet ratio of fecal particles, along with increased serum levels of creatinine, blood urea nitrogen, uric acid, and lipopolysaccharide. Mancozeb also markedly altered the gut microbiota structure, leading to an increased relative abundance of Lachnospiraceae_NK4A136_group (1.01-1.71-fold) and Ileibacterium (2.21-5.54-fold). Furthermore, mancozeb promoted the accumulation of acetylcholine, histamine release, and elevated levels of indole compounds in intestinal tissues. Transcriptomic analysis revealed upregulation of immune-related genes and downregulation of genes involved in drug metabolism. These results suggest that mancozeb exposure may affect gut microbiota composition, neurotransmitter and metabolite profiles, and the expression of functional genes related to immunity and detoxification. This study provides critical insights for the dietary health risk assessment of mancozeb.

Organisms adapt to fasting by remodeling energy metabolism across tissues. We identify geranylgeranyl pyrophosphate synthase (GGPPS) in white adipose tissue (WAT) as a fasting-responsive regulator that orchestrates lipolysis and branched-chain amino acid (BCAA) metabolism. In mice, fasting downregulates GGPPS, destabilizing the BCAA-catabolizing enzyme BCAT2 via enhanced ubiquitination. Mechanistically, GGPPS scaffolds the deubiquitinase OTUB1 and BCAT2 to stabilize BCAT2, promoting BCAA catabolism. Elevated BCAAs activate adipocyte lipolysis and enhance hepatic fatty acid oxidation. Adipose-specific deletion of Ggpps phenocopies fasting responses, including enhanced lipolysis and systemic BCAA elevation. Conversely, BCAA supplementation promotes adipose lipolysis, protects against diet-induced obesity, and facilitates hepatic oxidative adaptation. In fasting mice, BCAA supplementation enhanced weight loss efficiency and prevented aberrant hepatic lipid accumulation. These findings identify GGPPS as a potential central node linking amino acid and lipid metabolism during early fasting within a multi-organ metabolic circuit.

More than 300 million people globally have viral hepatitis. Predominantly caused by hepatitis A, B, and C viruses, viral hepatitis continues to confer high levels of morbidity and mortality despite the availability of vaccines and advances in therapies. Symptom severity varies during acute infection, with management focused on stabilization and serial monitoring to ensure resolution of liver injury. Infection with hepatitis A virus is generally self-limited, with low but increasing rates of mortality and hospitalization due to factors such as poorer baseline health status, older age, and coexisting liver disease. About 95% of acute hepatitis B cases in adults resolve spontaneously; treatment is rarely needed in acute infection. In contrast, only about 25% of acute hepatitis C cases resolve spontaneously, and immediate initiation of treatment at diagnosis is recommended. Screening for hepatitis B and C is recommended for all adults. Hepatocellular carcinoma screening is recommended for those with hepatitis B and cirrhosis or specific risk factors, and for those with hepatitis C and advanced liver fibrosis or cirrhosis. Treatment of hepatitis B involves assessment of specific factors to determine antiviral need; the goal of treatment is suppression of viral replication, thereby reducing risks of liver disease progression and hepatocellular carcinoma. Curative medications are available for hepatitis C.

Cancer cachexia is a devastating systemic syndrome characterized by progressive body weight loss and multi-organ dysfunction, yet the proteome-level mechanisms driving synchronized organ remodeling remain incompletely defined. Here, we applied large-scale data-independent acquisition (DIA) proteomics to a reproducible xenograft model using cachexia-inducing human neuroendocrine carcinoma cells (AkuNEC). Compared with non-implanted controls, AkuNEC-bearing mice developed severe wasting of the heart, liver, kidney, and skeletal muscle. Quantitative profiling revealed extensive multi-organ proteome remodeling, with xenobiotic metabolism emerging as a recurrently altered program across all tissues. This shared "chemical stress" signature was overlaid with distinct organ-specific alterations. The liver, heart, and kidney exhibited convergent suppression of mTORC1 signaling, with the liver displaying additional complex reprogramming involving interferon responses and fatty acid metabolism. In contrast, skeletal muscle showed unique stress features, with coagulation emerging as the most prominent signature alongside xenobiotic metabolism. These findings establish a comprehensive multi-organ proteomic framework for cachexia, identifying systemic remodeling of xenobiotic and endobiotic stress pathways as a unifying pathophysiological feature. This pan-organ alteration implies a fundamental compromise in the host's capacity to detoxify endobiotics and therapeutics, providing a molecular rationale for the unpredictable pharmacokinetics and heightened drug toxicity frequently complicating cachexia management.