Peptide-based drug candidates are often susceptible to enzymatic degradation in the gastrointestinal environment and biological matrices, while exhibiting limited intestinal permeability. In developing strategies to overcome these limitations, it is important to quantitatively evaluate the factors contributing to low systemic exposure and to understand the metabolic fate of peptides. This study utilized an integrated analytical approach combining quantitative analysis via liquid chromatography tandem-mass spectrometry (LC-MS/MS) and metabolite profiling via LC-quadrupole time-of-flight MS (LC-QTOF-MS) to characterize the metabolic fate and evaluate the oral dosing feasibility of WKYMVm (Wm), a formyl peptide receptor 2 agonist. A validated LC-MS/MS assay enabled quantification of Wm in mouse plasma with acceptable accuracy (95.2-115.7%) and precision (<8.6%). LC-QTOF-MS analysis identified eight metabolites (M1-M8) in mouse plasma, predominantly formed via sequential N-terminal cleavage with oxidative modifications. Plasma stability data were then simultaneously fitted to a structural parent-metabolite kinetic model, which identified the formation of M4 as the predominant pathway (fraction metabolized (fm) = 0.501), followed by M2 (fm = 0.262), indicating preferential cleavage at the Lys-Tyr bond. To predict oral dosing feasibility, Caco-2 bi-directional transport data were analyzed using a catenary model, while luminal degradation and hepatic metabolism were estimated using simulated gastric and intestinal fluids and mouse liver S9 fraction. When the experimentally obtained parameters were incorporated into simplified models, the results revealed limited intestinal and hepatic availability under the tested conditions. This integrated analytical and modeling approach identified the metabolic liabilities of Wm, informing the design of Wm analogs to enhance metabolic stability.

Obesity is a chronic, progressive disease defined by excess adiposity that can negatively affect a person's health. Approximately one billion people are living with obesity globally, though this estimate is shaped by the ongoing evolution in how the condition is measured and diagnosed. Although obesity is traditionally diagnosed by body mass index (BMI), it does not adequately assess body fat distribution, particularly visceral fat, which is specifically linked to an increased risk of poor metabolic health and cardiovascular disease. Other anthropometric measures such as waist circumference and waist-to-height ratio assess an indirect measure of visceral adiposity and are better predictors of cardiovascular risk than BMI alone. Obesity can increase the risk of developing type 2 diabetes, lipid dysfunction, and metabolic dysfunction-associated steatotic liver disease (MASLD), a condition in which the liver stores too much fat because the body's metabolism is not working properly. Around 75% of people with obesity have MASLD, with 12% to 40% progressing to metabolic dysfunction-associated steatohepatitis (MASH; the advanced form of MASLD) within 8-13 years. MASH and MASLD are largely underdiagnosed because someone affected by these conditions does not show symptoms. In this podcast, a patient and doctor specialising in obesity medicine share their perspective on the experience and best practice relating to the diagnosis of obesity, the need for screening of obesity complications, and the importance of the assessment and management of liver health as part of a holistic delivery of obesity care.Podcast available for this article. Podcast (MP4 156963 kb).

BackgroundMidlife obesity is considered one of the top modifiable risk factors for dementia and Alzheimer's disease (AD). However, body mass index (BMI) on its own does not fully represent obesity-associated risks and it is crucial to disentangle the role of body adiposity and its localization.ObjectiveTo investigate the relationship of MRI-derived body adiposity metrics with AD-related pathology at midlife.MethodsNinety-seven cognitively normal midlife individuals underwent brain amyloid and tau PET, body MRI, and metabolic and cognitive assessments. Key measures included hepatic fat fraction, visceral (VAT) and subcutaneous adipose tissue (SAT) volumes, and thigh muscle and adiposity. The correlation between adiposity/metabolic measurements and amyloid/tau pathologies was investigated.ResultsThe average age of participants was 49.8 years, 65.3% were female and 53.6% had obesity. Amyloid PET burden in Centiloids correlated with VAT (rho = 0.36, p = 0.002), BMI (rho = 0.33, p = 0.002), SAT (rho = 0.33, p = 0.002), and insulin resistance (IR) (rho = 0.34, p = 0.003) in females and Whites, lower high-density lipoprotein (HDL) cholesterol (rho = -0.36, p = 0.002) irrespective of sex and race, and lower MMSE scores (rho = -0.57, p = 0.043) in only in African-Americans, after correction for age, sex, and education. There was no evidence that HDL nor IR mediated VAT-related amyloid. VAT/SAT ratio was significantly associated with mean cortical tau SUVR (β = 0.138, p = 0.030) after adjustment for age, sex, education, and amyloid.ConclusionsAmong fat depots in our study, visceral fat was more strongly correlated to amyloid pathology, and this association is present even independent from BMI. Also, higher visceral compared to subcutaneous fat is related to higher tau pathology.

Purpose We aim to identify possible factors affecting liver regeneration in donors, liver graft hypertrophy and spleen shrinkage in recipients. We aim also to assess the differences between donors and recipients.Methods and methodsTwenty-seven donors and 27 recipients were included. We collected anthropometric and bio-humoral parameters considered in the preoperative setting, just after the surgery, after 7 days, and after 30 days from the surgery. We calculated liver regeneration rates (RR%), regeneration velocities (RV) and spleen reduction rates (SRR%) at 2-3 months, 12 months, and 24 months. Body composition parameters were calculated.ResultsIn right lobe donors we found a mean RR of 118,70%, 146,33% and 154,88%. The mean RV was 7,94 cc/s, 0,61 cc/s, and 0,24 cc/s.For right lobe recipients the mean RR was 72,61%, 68,31%, and 79,69%. The mean RV was 9,74 cc/s, 0,19 cc/s, and 0,16 cc/s. The mean spleen reduction rate was 1,38%, 11,36%, and 10,80%.The regeneration rates were higher in donors with statistically significant differences. The recipients have a growth peak within 2-3 months, while donors at 12 months.In donors, the residual liver volume after transplantation and pre-transplant liver function are negatively correlated with regeneration. In recipients, we observed negative correlations with hematological and nutritional parameters. Liver regeneration goes parallel with the volumetric reduction of the spleen. A greater muscle mass and improved muscle quality are major stimuli to the regeneration process.ConclusionThis study emphasizes how sophisticated and versatile liver regeneration is, influenced not only by the remaining liver or graft, but also by the metabolic environment of the host.

Adzuki bean saponin (ABS) has shown therapeutic potential in improving obesity, but its underlying mechanism and active saponin components are still unknown. This research estimated the anti-obesity effect of ABS in obesity using a mouse model. Advanced research methods such as spatial metabolomics, transcriptomics, proteomics, serum pharmacology, and network pharmacology were employed to study the molecular mechanisms of ABS and its bioacitve saponins. ABS achieved anti-obesity effects by improving HFD-induced body weight gain, abnormal lipid profiles and liver dysfunction liver lipid accumulation, and increased adipose tissue. Liver spatial metabolomics suggested a potential involvement of ABS in modulating lipid metabolism via alterations in the arachidonic acid pathway. The results of transcriptomics, proteomics, and immunohistochemistry indicated that ABS could achieve anti-obesity effects through activating the Wnt/β-catenin signaling pathway. Based on our previous study and serum pharmacology analysis revealed that soyasaponin Ba is a key saponin in ABS that achieves anti-obesity effects. Similarly, soyasaponin Ba can achieve anti-obesity effects by activating Wnt/β-catenin signaling. ABS and its active saponin Soyasaponin Ba can achieve anti-obesity effect by up-regulation of the Wnt/β-catenin signaling pathway. This understanding of this mechanism provides valuable insights for developing drugs or health supplements based on adzuki beans to combat obesity.

This study aims to generate synthetic-MRI(synMRI) for abdominal patients undergoing radiotherapy using Deep Inspiration Breath Hold(DIBH) on a conventional Linac.&#xD;Methods: XCAT phantom and patient data were used to develop and test a patient-specific model to generate synMRI. During patient simulation, 6 DIBH-MRIs and 1 DIBH-CT were acquired. One DIBH-MRI was set as the reference MRI(refMRI) and Principal Component Analysis(PCA) was performed using five Deformation Field Maps(DFMs) by applying deformable image registration between refMRI and the other DIBH-MRIs. PCA eigenvalues were sampled 1,000 times to generate new deformations applied to a synthetic-CT with the same anatomic conditions as refMRI. A convolutional neural network was trained to predict the eigenvalues corresponding to on-board conditions from a CBCT to generate a final synMRI with the new DFM. Four XCAT scenarios simulated changes from simulation to treatment. The model was evaluated using mean-absolute-error(MAE) and root-mean-square-error(RMSE), and the image quality was evaluated by structure-similarity-index metric(SSIM) and normalized RMSE(nRMSE). The accuracy of the predicted target volume for XCAT and fiducial clips for each patient was analyzed using center-of-mass-shift(COMS) between on-board conditions (ground-truth synMRI for XCAT and CBCT for patients) and the predicted synMRI. The liver dome difference was also evaluated.&#xD;Results: Model performance yielded an MAE of 0.11±0.02 for XCAT, and 0.15±0.01, 0.15±0.02, 0.10±0.03, 0.12±0.09 for four patients, respectively. RMSE values were 0.11±0.07, 0.16±0.03, 0.19±0.01, 0.13±0.07, and 0.14±0.04. For image quality, SSIM values were 0.999±0.001 for XCAT, and 0.998±0.001, 0.995±0.002, 0.996±0.001, 0.998±0.001 for the patients. The nRMSE were 0.10±0.06, 0.58±0.03, 0.87±0.06, 0.39±0.04, and 0.35±0.02, respectively. For XCAT, the liver dome difference and tumor COMS were <0.5 mm in all scenarios. For all patients, liver dome differences were <1 mm, and fiducial COMSs were <1.2 mm.&#xD;Conclusions: The novel method generates synMRI using on-board conditions in DIBH abdominal treatments performed on a conventional LINAC.&#xD.

Recent advances in lipid nanoparticle (LNP) delivery systems have enabled the development of mRNA vaccines and oligonucleotide therapeutics. Reliable hepatic in vitro systems are essential for the evaluation of the biodegradation of LNP components. SM-102 is an important ionizable lipid used in Moderna's LNP-mRNA vaccine. In this study, we compared the metabolism of SM-102 using 3 in vitro hepatic models, including liver microsomes, 2D hepatocyte cultures, and 3D liver spheroids prepared from primary rodent hepatocytes and HepG2 cells. An ultra-high-performance liquid chromatography high-resolution mass spectrometry method was developed to detect and characterize SM-102 metabolites. A 2,4-dinitrophenylhydrazine derivatization method was developed to further capture and characterize the potential carbonyl products. Following incubation of SM-102 with liver microsomes from human, monkey, dog, rat, and mouse, 7 metabolites were identified, with oxidation to carbonyl-containing metabolites representing the predominant pathway. In mouse 2D hepatocytes and 3D liver spheroids, 6 metabolites were detected, and the major metabolic pathways included N-dealkylation (M6), acetylation (M7), and hydrolysis (M8). In rat 2D hepatocyte cultures, 6 metabolites were detected, with M6 and M8 as the major metabolites. In contrast, β-oxidation-related metabolites (M10 and M11) were the major metabolites detected in rat 3D hepatocyte spheroids. Comparable metabolic profiles of SM-102 were observed in HepG2 2D cultures and 3D spheroids, with hydrolysis and N-dealkylation as the predominant metabolic pathways. Taken together, this study characterized the metabolic profiles of SM-102 across 3 in vitro hepatic models, and comparable metabolic profiles were observed between 2D hepatocytes and 3D spheroids prepared from primary mouse hepatocytes and HepG2 cells. These findings provide useful information for selecting appropriate in vitro models to ensure the reliable evaluation of ionizable lipid biodegradation. SIGNIFICANCE STATEMENT: This study compared the metabolic profiles of SM-102 across 3 hepatic models. 2D hepatocyte and 3D liver spheroid identified hydrolysis, N-dealkylation, and β-oxidation as major metabolic pathways for SM-102. In contrast, liver microsomes predominantly drove the oxidative metabolism. These findings highlight the necessity of cell-based models for the reliable evaluation of ionizable lipid biodegradation.

Anticoagulant rodenticides (ARs), molluscicides, and neonicotinoids are widely used biocides for plant protection and pest control and are considered a potential health risk to non-target wildlife. Here, we investigate these compounds in convenience samples derived from 100 European hedgehogs (Erinaceus europaeus) and 181 red foxes (Vulpes vulpes) from England (2013 to 2020), screening liver tissues using liquid chromatography-tandem mass spectrometry. Whilst no molluscicides or neonicotinoids were detected, except for low level clothianidin in a single fox, ARs were found in 62% of hedgehog and 90% of fox samples at variable concentrations (0.05-3430 ng/g). In 83% of animals with detectable ARs, more than one AR compound was found, consistent with multiple, cumulative exposures. Additionally, 25% of hedgehog and 64% of fox samples contained summed AR concentrations above the commonly used reference value of 100 ng/g for potential poisoning or regulatory concern. There was no significant association with geographical region or sex (for fox or hedgehog), or with age, location, habitat type, or the assigned category of cause of death (for hedgehog). Despite the high AR levels detected, there was no evidence of acute haemorrhage indicative of AR toxicosis in hedgehogs. However, since hedgehogs are of conservation concern in Great Britain, further investigation of potential adverse health impacts of ARs and other pesticides is warranted. Our study findings indicate widespread AR exposure among the sampled animals and support further structured monitoring of these omnivorous non-target mammals. Within our convenience sample, we found no evidence of reduced AR exposure following the introduction of the UK rodenticide stewardship scheme in 2016.

Irisin is a myokine that enhances insulin secretion and β cell viability in both rodent and human β cells. Here, we investigated whether irisin preserves β cell functional mass in vivo in diabetic mice and ex vivo in human pancreatic islets from subjects with type 2 diabetes (T2D). In mice made diabetic by a high-fat diet and streptozotocin, irisin administration improved glycemic homeostasis by increasing islet insulin content and glucose-stimulated insulin secretion while also markedly stimulating β cell proliferation. In islets from T2D subjects, which typically display substantial structural and functional impairments, irisin restored insulin content and glucose-stimulated insulin secretion. Mechanistic studies in INS-1E cells showed that chronic exposure to high glucose blunted glucose-triggered cytoplasmic calcium increases, essential for insulin secretion. Under these glucotoxic conditions, while irisin was unable to activate CREB and AKT, it induced AMPK-mTORC1-S6K-dependent mobilization of endoplasmic reticulum calcium stores, resulting in enhancement of insulin secretion. Overall, these findings highlight irisin's multifaceted ability to counteract β cell failure in experimental and human T2D.

Gallstones are a common digestive disorder whose development is influenced by abnormal liver function, metabolic disorders, and inflammatory states. The albumin-bilirubin (ALBI) score is a comprehensive indicator for assessing liver function and has been widely applied in liver cancer and chronic liver diseases; however, its association with gallstones remains unclear. This study analyzed NHANES data from 2017 to 2020, including 6332 adult participants. Survey-weighted multivariable logistic regression was performed to assess the association between ALBI and gallstone prevalence. Furthermore, the dose-response relationship was examined using restricted cubic spline (RCS) modeling. Subgroup analyses were conducted to assess the robustness of this association across different demographic and clinical strata. In addition, an exploratory mediation analysis was performed to examine whether body mass index (BMI) statistically accounted for part of the observed association between ALBI and gallstone prevalence. Multivariable logistic regression indicated that higher ALBI was associated with higher odds of gallstone prevalence (OR = 2.670, 95% CI: 1.778 - 4.008, P = 0.001). RCS analysis indicated that the relationship between ALBI and gallstones is approximately linear (P-overall < 0.0001, P-nonlinearity = 0.7779). The results of the subgroup analyses were consistent and robust. Exploratory mediation analysis suggested that BMI statistically accounted for 23.95% of the observed association between ALBI and gallstone prevalence in the mediation model. Higher ALBI levels were associated with a higher prevalence of gallstones, showing an approximately linear dose-response pattern. BMI statistically accounted for part of this association in exploratory mediation models, but this finding should not be interpreted as evidence of causal mediation given the cross-sectional design. Because gallstone status was self-reported, these findings should be interpreted with caution. Prospective studies are warranted to clarify temporal relationships, validate these associations, and determine whether ALBI provides clinically meaningful information beyond established gallstone risk factors.