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This study examined how different photoperiods affect net energy partitioning and explored the mechanisms via blood biochemistry, gut microbiota, and fecal metabolites. Twelve healthy crossbred pigs (47.7 ± 7.5 kg) were randomly allocated to two groups and subjected to a self-controlled crossover design. Following an 8-day baseline under a normal photoperiod (12L:12D, 12 h light:12 h dark), pigs were assigned to two photoperiod treatment groups: prolonged photoperiod (18L:6D, 18 h light:6 h dark; P group) and shortened photoperiod (6L:18D, 6 h light:18 h dark; S group). Measurements during the baseline (12L:12D) and treatment phases are designated as N1/P (for the P group) and N2/S (for the S group), respectively. The treatment periods were interspersed with the baseline 12L:12D photoperiod and repeated six times. It was observed that, compared to N2, shortened photoperiod (S) had significantly higher net energy deposition, net energy for protein deposition, and net energy for fat deposition ( Show less

Apolipoprotein A-IV amyloidosis (AApoA-Ⅳ) is a rare subtype of cardiac amyloidosis (CA) and is often overlooked because of its clinical similarity to transthyretin (ATTR) CA. An 82-year-old man presented with heart failure with preserved ejection fraction. His clinical features, including grade 1 uptake on Although the pathologic significance of the unique histologic features remains unclear, they may represent distinguishing characteristics of AApoA-IV amyloidosis. AApoA-IV CA typically presents with elderly-onset heart failure with preserved ejection fraction and may show positive uptake on bone scintigraphy. Differentiation from ATTR CA is possible based on characteristic histopathologic findings and mass spectrometry-based proteomic analysis. Show less

Skin aging arises from intrinsic processes and extrinsic insults (e.g., ultraviolet exposure and oxidative stress). Mesenchymal stromal cell (MSC)-derived secretome offers a cell-free approach to skin regeneration. Wharton's jelly-derived MSCs (WJ-MSCs) may outperform adipose-derived (AD-MSCs) and bone marrow-derived MSCs (BM-MSCs). Secretomes from WJ-MSCs, AD-MSCs, and BM-MSCs were compared in vitro for human dermal fibroblast proliferation, scratch-wound closure, extracellular-matrix (ECM) remodeling, and type I procollagen secretion. Anti-inflammatory and antioxidant activities were assessed by IL-6, IL-1β, TNF-α, COX-2 and intracellular reactive oxygen species (ROS). Antibody arrays profiled secreted factors. An exploratory, single-arm human pilot ( The WJ-MSC secretome increased fibroblast proliferation, ECM remodeling, and type I procollagen, and reduced cytokines and ROS, exceeding the effects of AD-MSC and BM-MSC secretomes. Profiling highlighted apolipoprotein A4 (ApoA4) and SERPINH1 as enriched, functionally active mediators; recombinant ApoA4 and SERPINH1 enhanced fibroblast activity, collagen-related readouts, and accelerated in vitro wound closure. In the pilot study, within-subject increases in instrument-derived hydration and elasticity were observed over one week (paired tests). No treatment-related adverse events were noted. Patch testing showed no irritation (ICDRG scores all 0; non-irritant classification). The WJ-MSC secretome demonstrated consistent in-vitro pro-regenerative, anti-inflammatory, and antioxidant activities, with ApoA4 and SERPINH1 as candidate mediators. Human findings are preliminary/exploratory and suggest potential short-term benefits that require confirmation in adequately powered, controlled trials. Show less

IntroductionAging and metabolic disease enhance inhaled particulate toxicity. Nanoparticles (NPs) are rapidly coated with biomolecules forming a biocorona (BC), upon entering the body and may contribute to the susceptibility. Aging and metabolic syndrome (MetS) are progressive conditions resulting in biomolecule alterations over time potentially influencing susceptibility. We hypothesize NP-biomolecule interactions are altered during aging and throughout MetS progression.MethodsC57BL/6J mice at 6 weeks of age were fed a healthy diet or a high-fat western diet. BALF was collected after 2, 4, 8, 12, 16, 20 or 24 weeks on diets. NP-biomolecules interactions were compared between healthy and MetS to determine age- and disease progression-related BC variations (proteins and lipids).ResultsUnique BCs were determined to form at each time point indicative of aging for the healthy and aging and disease progression for the MetS. Comparisons between healthy and MetS BCs at each time demonstrated distinct biomolecule interactions attributable to disease. Comparisons determined both unique protein and lipid content as well as quantitative differences. Proteins such as apolipoprotein A-IV, complement C3 and lipids such as PE (37:5), PE (O-38:5), PE (P-38:4), PC(40:7), PC(39:0), and PC(O-40:0) were identified on the MetS BC suggesting disease progression modifications. Proteins such as pulmonary surfactant protein A, fibrinogen alpha-chain and lipids such as CE (19:0)-NH4, DG (36:7), and DG (35:0)_C18:0 were increasingly present in the healthy BC over time, suggesting age-related interactions.DiscussionOverall, unique BCs were identified demonstrating the impact of age and disease progression on BC formation which will aid in understanding initial pulmonary NP-biomolecular interactions potentially contributing to susceptibility. Show less

Diet-based modulation of the gut microbiota has emerged as a promising strategy to alleviate obesity and its related complications. Our previous study demonstrated that polysaccharide derived from Cordyceps militaris (CMP) exerts anti-obesity effects, yet the specific mechanism linking gut microbiota to its metabolic impact remains unclear. Herein, we utilized murine models with distinct gut microbial profiles created via antibiotic cocktails to investigate these mechanisms. The protective effects of CMP against high-fat diet (HFD)-induced obesity and associated metabolic disturbances were substantially impaired in mice depleted of neomycin-sensitive gut bacteria. Metagenomic analyses further established that CMP required these bacteria to restore gut microbial homeostasis. Notably, we observed that CMP elevated hepatic levels of brassicasterol in a manner dependent on neomycin-sensitive gut bacteria. Brassicasterol treatment alone replicated the anti-obesity effects of CMP, as indicated by reduced body weight gain, improved lipid and glucose metabolism, and decreased inflammation. Through transcriptomic and functional analyses, we identified hepatic Apoa4 as a key downstream effector of brassicasterol. Our results indicated that brassicasterol upregulated Apoa4, facilitating lipid transport and suppressing inflammation both in vitro and in vivo. Collectively, our findings indicate that CMP exerts its anti-obesity effects through a neomycin-sensitive gut bacteria-brassicasterol-Apoa4 pathway. This work expands the mechanistic understanding of CMP and highlights a novel microbiota-metabolite-host regulatory axis for dietary intervention in metabolic disorders. Show less

'Small extracellular vesicles (sEVs) are nanosized, membrane-enclosed sacs released by diverse cell types. They play a critical role in cell-cell communication through their cargo, which includes a wide range of proteins, lipids, and nucleic acids. Physiologically, sEVs circulate in various body fluids such as blood, urine, and saliva, making them accessible for diagnostic via non-invasive isolation techniques. Recent advances in high-throughput proteomics have significantly enhanced our ability to characterize the protein content of sEVs. Importantly, multiple studies on human fluids have identified specific protein markers across different cancer types, encompassing molecules involved in inflammation, cellular adhesion, immunity, and lipoprotein regulation. Interestingly, some of these proteins are consistently detected across multiple cancer types and sample sources, suggesting the existence of a shared "oncogenic signature" that may be transferred via sEVs. Among body fluids, urine and saliva are particularly promising for easy, non-invasive diagnostics. However, these sample types remain underexplored as compared to the serum, leaving substantial opportunities for future research. Taken together, these findings position sEVs as a powerful tool with significant potential for advancing precision cancer care. SIGNIFICANCE: Living cells release nanosized membrane-enclosed vesicles called small extracellular vesicles (sEVs) into the extracellular environment. sEVs contain protein cargo molecules that critically take part in cell-cell communications. Quantitative proteomics identified potential sEV associated biomarkers for early cancer diagnosis and therapy. sEV Proteins associated with cell adhesion and inflammation, lipoproteins and immunoglobulins are potential molecules that were majorly identified. Interestingly, some of these proteins such as APOA4, SAA4, ITIH4, SERPINC1 and VWF were consistently identified across multiple cancer types and sample sources, highlighting their potential as future biomarkers. Show less

Alterations in the gut microbiome and a "leaky" gut are associated with Parkinson's disease (PD), which implies the prospect of rebalancing via dietary intervention. Here, we investigate the impact of a diet rich in resistant starch on the gut microbiome through a multi-omics approach. We conducted a randomized, controlled trial with short-term and long-term phases involving 74 PD patients of three groups: conventional diet, supplementation with resistant starch, and high-fibre diet. Our findings reveal associations between dietary patterns and changes in the gut microbiome's taxonomic composition, functional potential, metabolic activity, and host inflammatory proteome response. Resistant starch supplementation led to an increase in Faecalibacterium species and short-chain fatty acids alongside a reduction in opportunistic pathogens. Long-term supplementation also increased blood APOA4 and HSPA5 and reduced symptoms of PD. Our study highlights the potential of dietary interventions to modulate the gut microbiome and improve the quality of life for PD patients. Show less

Depression is a heterogeneous psychiatric disorder with limited treatment efficacy, as 30-50% of patients exhibit inadequate responses to conventional monoaminergic antidepressants. Rhein, a bioactive anthraquinone derived from Rheum palmatum, exhibits rapid and sustained antidepressant effects in both acute and chronic social defeat stress (CSDS) mouse models. Using quantitative proteomics on prefrontal cortex (PFC) samples from control, CSDS, Rhein-treated, and imipramine-treated cohorts, we identified differentially expressed proteins that revealed Rhein's multi-target regulatory profile. Functional enrichment and clustering analyses indicated that Rhein predominantly restores dysregulated pathways related to lipid metabolism, ribosomal translation, mitochondrial and endoplasmic reticulum (ER) function, and synaptic plasticity, forming a coherent mechanistic axis underlying its therapeutic effects. Comparative analysis with imipramine-treated mice further highlighted Rhein's distinct capacity to modulate organelle homeostasis and synaptic remodeling with greater breadth. Parallel reaction monitoring (PRM) and Western Blotting validated key proteins involved in mitochondrial functions (BNIP1, PISD, MRPL42, MRPS30, LRBA, IGHM), ER homeostasis (ACBD5, APOA4, RPL14), and synaptic plasticity (HDAC1, FAM3C, SSU72). These molecular findings suggest that Rhein exerts its antidepressant effects by restoring the functional integrity of mitochondria and the ER, thereby reprogramming synaptic plasticity. We inferred that this organelle-centered regulation further reinforces its potent modulation through multiple mechanisms and signaling pathways of synaptic plasticity, enabling Rhein to exert antidepressant effects through a coordinated, multi-layered mechanism. Collectively, our findings provide a systems-level mechanistic framework for Rhein's antidepressant efficacy and support its potential as a multi-pathway natural therapeutic, particularly for metabolic subtypes of depression. Show less

Breast cancer is the leading cause of cancer-related mortality in African American (AA) women. In this study we evaluated the serum proteomic profile of AA women with breast cancer using an integrated proteomic framework with multivariate pattern analysis. Using 2D-DIGE, thousands of serum protein spots were detected across 33 gels; 46 spots met criteria for presence, statistical significance, and differential expression. Proteins from the spots were identified by MALDI-TOF/TOF and matched in curated databases, highlighting serum biomarkers including ceruloplasmin, alpha-2-macroglobulin, complement component C3 and C6, alpha-1-antitrypsin, alpha-1B-glycoprotein, alpha-2-HS-glycoprotein and haptoglobin-related protein. LC-MS/MS analysis revealed 163 differentiating peptides after imputing and filtering 286 peptides. These were evaluated using cumulative distribution function (CDF) analysis, a nonparametric method suited for limited sample sizes. Peptide patterns were explored with Random Forest, showing concordance with CDF. The model achieved an AUC of 0.85 at the peptide level. This workflow identified differentiating proteins (CERU, A2MG, CO3, VTDB, HEMO, APOB, APOA4, CFAH, CO4A, AACT, K1C10, ITIH2, ITIH4), highlighting CERU, A2MG, and CO3 with overexpression and reproducible identification across platforms. We present an integrated, non-invasive serum protein biomarker signature panel specific to AA women, through reproducible proteomic sensor framework to support early detection and breast cancer prevention. Show less

Imperfect first-trimester screening for hypertensive disorders of pregnancy (HDP) means many high-risk women miss the window for preventive aspirin, and the biological heterogeneity of HDPs is overlooked. This study aimed to leverage first-trimester serum proteomics to create a more precise tool for predicting preeclampsia (PE) and differentiating it from other HDPs. A prospective nested case-control study ( Show less

Lung adenocarcinoma (LUAD) is the most common subtype of lung cancer and is difficult to distinguish from benign pulmonary nodules (BPNs), particularly at early stages. Extracellular vesicles (EVs) represent a promising source of biomarkers for the diagnosis of malignant pulmonary nodules. This study aimed to identify robust and clinically relevant EV-based protein biomarkers via isolation with EXODUS, a system that enables efficient direct capture of plasma EVs, followed by data-independent acquisition mass spectrometry (DIA-MS) for in-depth proteomic profiling. A total of 1383 proteins were identified from the plasma EVs obtained from 25 individuals (10 BPN and 15 early stage LUAD), while dysregulated protein signatures were revealed through differential expression analysis. Machine learning algorithms incorporating demographic variables, imaging features, EV protein profiles, and conventional tumor markers were applied to select diagnostic candidates. Random forest analysis revealed two upregulated proteins, NTN3 and APOA4, as promising biomarkers. Subsequently, their diagnostic performance and net clinical benefits were validated in an independent EV cohort (6 LUAD and 6 BPN) using ELISAs and decision curve analysis. In summary, we present an integrated pipeline that combines EXODUS-based isolation, DIA-MS, and machine learning to detect markers from plasma EVs for distinguishing early stage lung cancer from benign nodules. Show less

Elite training induces profound physiological adaptations, yet whether these changes manifest as stable circulating proteomes remains unclear. This study characterized serum proteomic profiles in male and female Olympic-level athletes to identify biomarkers associated with long-term endurance and strength training. Cross-sectional study in Olympic-level athletes and sedentary controls. Resting serum samples were collected from male and female marathon runners and weightlifters (with 5-15 years of training), alongside age- and sex-matched sedentary individuals. Proteomic profiling was performed using tandem mass spectrometry. Data were processed with MaxQuant and analyzed using Perseus. Selected proteins were confirmed using antibody-based assays. Among 301 identified protein groups, 36 showed significant differences between groups. Apolipoprotein A-IV (APOA4) was elevated in athletes, particularly marathoners, suggesting cardiovascular adaptation to endurance training. Fibronectin 1 (FN1) was reduced in weightlifters, consistent with vascular remodeling associated with resistance training. Marathoners exhibited higher levels of von Willebrand factor (VWF) and glycosylphosphatidylinositol-specific phospholipase D1 (GPLD1), and lower levels of galectin-3-binding protein (LGAS3BP) and leucine-rich alpha-2-glycoprotein 1 (LRG1), indicating immunomodulatory effects of oxidative training. Weightlifters showed reduced levels of GPLD1 and extracellular matrix protein 1 (ECM1), reflecting distinct remodeling pathways. FN1, APOA4, VWF, LGALS3BP and ECM1 levels were further confirmed. Endurance and resistance training elicit modality-specific serum proteomic adaptations that reflect vascular, endothelial, and hemostatic remodeling. These molecular signatures, observed in both sexes, highlight stable changes induced by chronic training and may inform cardiovascular prevention strategies and evidence-based approaches in sports science to optimize training and performance. Show less

Rising global temperatures lead to a continuous increase in the frequency and intensity of extreme weather events, such as droughts and floods, posing serious threats to terrestrial homeotherms. However, adaptive changes in respiratory metabolism and molecular mechanisms in lung tissues of small mammals under extreme water shortage conditions remain unclear. This study hypothesized that small desert mammals can adapt to extreme water shortage environments by regulating the plasticity of lung tissue gene expression and respiratory metabolism. Using 29 wild-caught Siberian jerboas ( Show less

Fibrillary glomerulonephritis is a rare cause of proteinuric kidney disease characterized by Congo red-negative fibrillary deposits and typically shows DNAJB9 positivity on immunohistochemistry. Amyloidosis is defined by Congo red positivity and can be typed by laser microdissection-tandem mass spectrometry when routine studies are inconclusive. We report the case of a 64-year-old man with proteinuria and declining kidney function whose kidney biopsy showed DNAJB9-positive fibrillary glomerulonephritis in glomeruli, but Congo red-positive deposits confined to the medulla were DNAJB9 negative. Laser microdissection-tandem mass spectrometry of the medullary deposits identified apolipoprotein A-IV amyloidosis, establishing concurrent fibrillary glomerulonephritis and apolipoprotein A-IV amyloidosis in the same biopsy. Apolipoprotein A-IV amyloidosis is often medullary predominant and, in rare hereditary forms related to autosomal dominant Show less

Significant interindividual variability in radiosensitivity poses a major challenge to conventional radiation protection and radiotherapy. Current prediction strategies relying on DNA damage or genomic analysis have inherent limitations, underscoring the need for minimally invasive serum biomarkers. While serum apolipoproteins are crucial regulators of lipid transport, metabolism, and cellular stress response, their role as biomarkers for radiosensitivity remains largely unexplored. A 7.3 Gy ⁶⁰Co γ-ray whole-body irradiation mouse model (with training and independent validation cohorts) was established to assess individual radiosensitivity. Pre-irradiation peripheral serum samples underwent high-throughput proteomics analysis to identify differential proteins (DEPs) linked to 30-day post-irradiation survival. KEGG and GO enrichment analyses were conducted to characterize DEP-associated pathways. An XGBoost machine learning model was built using candidate biomarkers, with SHAP analysis to define their predictive contributions; Cox proportional hazards and Pearson correlation analyses were applied to evaluate survival associations. DIA-based proteomics identified 580 DEPs in the training cohort and 449 in the validation cohort. KEGG and GO enrichment analyses confirmed that these DEPs were predominantly enriched in the cholesterol metabolism and reverse cholesterol transport pathways. The predictive model based on an apolipoprotein panel (ApoA1/ApoA2/ApoA4), established using the XGBoost algorithm, exhibited exceptional performance in the training cohort (AUC = 1) and maintained robust generalizability in an independent validation cohort (AUC = 0.833). Compared with non-survivors, survivors exhibited significantly elevated serum levels of ApoA1 and ApoA2 but markedly reduced levels of ApoA4. Cox proportional hazards regression analysis established ApoA1 and ApoA2 as independent protective factors, whereas high ApoA4 expression was an adverse prognostic indicator. Notably, ApoA4 levels also demonstrated a strong negative correlation with post-irradiation survival time. The serum apolipoprotein profile (ApoA1/ApoA2/ApoA4) serves not only as a promising minimally invasive biomarker for predicting individual radiosensitivity in mice but also reveals a critical link between the cholesterol metabolic pathway and radiation response. This finding lays a theoretical foundation for translating predictive, cholesterol metabolism-related biomarkers to support radiation response assessments. Given the limitations of animal models, subsequent studies are required to validate the clinical applicability of this panel in human cohorts, with the aim of offering an effective tool for personalized radiation protection and precise radiotherapy. The online version contains supplementary material available at 10.1186/s12944-026-02868-8. Show less

Current infant formulas lack the native multilayer structure of breast milk fat globule membrane (MFGM), impacting lipid digestion. In this study, the inner layer material and concentration of the biomimetic fat globule membrane were optimized by comparing particle size, Zeta-potential and interface protein load. It was found that compared with sodium caseinate (CN) and whey protein (WP), when the lactoferrin (LF) concentration was 2 %, the particle size was lower (277.85 ± 6.15 nm) and Zeta-potential value was higher (19.67 ± 1.27 mv). Using milk phospholipid (MPL) as the outer layer material, when the MPL concentration was 2 %, the emulsion had a smaller particle size (291.33 ± 1.15 nm) and a better stability (10.22 ± 0.62 %). Therefore, the biomimetic multilayer membrane was constructed by electrostatic layer-by-layer deposition of 2 % LF and 2 % MPL. Combining Fluorescence and Fourier transform infrared spectroscopy (FTIR), the interaction between LF and MPL molecules in the LF-MPL multilayer structure is primarily a spontaneous, endothermic process driven by hydrophobic forces, exhibited superior stability (except thermal stability) than LF monolayer membrane. The results of in vitro digestion showed that compared with LF, WP and WP-MPL emulsions, LF-MPL emulsions had the highest free fatty acid (FFA) release rate of 69.97 %. LF-MPL enhanced gastric stability and promoted intestinal lipolysis and improved the degree of lipid digestion. In addition, LF-MPL promoted the absorption and utilization of triglyceride (TAG) in cells and animals, and secretion and upregulated lipid absorption genes (FATP4, DGAT1, APOB, APOA4, MTTP). These findings demonstrate that biomimetic LF-MPL multilayers improve lipid digestion, absorption, and bioavailability, providing a theoretical basis for designing more breast milk-like infant formulas. Show less

Fatigue persists as a dominant and debilitating phenomenon in long-COVID, yet its underlying biological mechanisms remain unclear. While inflammatory variables tend to normalize within months post-infection, fatigue continues to significantly impact quality of life. Understanding whether specific biomarkers associate with long-COVID fatigue could shed light on pathophysiological mechanisms and potential therapeutic targets. In this single-center, cross-sectional controlled study, we enrolled 48 individuals with long-COVID (according to NICE criteria) and 48 age- and sex-matched recovered controls with prior SARS-CoV-2 infection but no persistent symptoms. We carefully excluded all subjects with other diseases or conditions that could influence fatigue levels. Fatigue severity was assessed using three validated instruments: Fatigue Visual Analog Scale (fVAS), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), and SF-36 vitality subscale. Blood samples were analyzed for pro-inflammatory markers (CRP, TNF-α, IL-6, IL-1β) and biomarkers associated with cellular stress responses and neuroprotection (HSP90α, APOA4, Serpin F1/PEDF, Hemopexin). Anti-nuclear antibodies (ANA) were tested to assess potential autoimmune mechanisms. Depression was assessed using the Hospital Anxiety and Depression Scale, Depression Subscale (HADS-D). Long-COVID patients demonstrated significantly higher fatigue severity across all instruments compared to recovered controls: fVAS median scores 63 versus 5 (p < 0.001), FACIT-F scores 21.5 versus 49 (p < 0.001), and SF-36 vitality scores 25 versus 72.5 (p < 0.001). Depression scores were also significantly elevated in long-COVID cases. However, none of the measured biomarkers differed significantly between groups: HSP90α, Serpin F1, Hemopexin, APOA4, and CRP showed no differences, while TNF-α and IL-6 showed only tendencies toward higher levels in long-COVID (p = 0.07 and p = 0.07, respectively). IL-1β concentrations were in most cases below the lower limit of detection and were excluded from further analysis. ANA positivity was 10.4% in cases versus 4.2% in controls (p = 0.38) and did not influence fatigue levels. Multivariable regression analysis revealed no significant associations between biomarkers and fatigue severity. Fatigue in long-COVID represents severe, persistent disability comparable to observations in chronic inflammatory diseases and chronic fatigue syndrome but is not associated with traditional inflammatory biomarkers or cellular stress response proteins measured in peripheral blood. The absence of biomarker associations suggests that long-COVID fatigue may involve more complex mechanisms, potentially including persistent neuro-immune dysregulation, epigenetic changes, or pathophysiological processes not reflected in systemic biomarker concentrations including neurobiological mechanisms such as altered predictive processing and central nervous system-confined neuroinflammation. These findings highlight the need for alternative approaches to understanding and treating long-COVID fatigue beyond conventional inflammatory paradigms. Show less

Modifying nanomedicines with targeting ligands represents an encouraging strategy for active tumor targeting, but its clinical failure underscores ongoing challenges. Herein, a series of liposomes with different targeting ligands (e.g., PEGylation, folic acid, mannose, RGD peptide, and melittin) were rationally designed to investigate the principles and mechanisms governing tumor targeting and penetration profiles. In primary and lung metastatic breast cancer models, these liposomes exhibited a systematic tendency of intratumor distribution, with melittin-modified liposomes showing optimal tumor targeting and therapeutic performance. Further studies revealed that the ligand modifications in liposomes could modulate the composition of their protein corona, particularly the level of Apolipoprotein A4 (ApoA4), which, in turn, influenced tumor targeting and intratumor distribution, ultimately affecting the therapeutic outcome of tumor inhibition and survival prolongation. This research provided a distinct correlation between ligand modification of liposomes and their Show less

Ultracentrifugation (UC) has long been considered the "gold standard" for extracellular vesicle (EV) isolation. However, due to its drawbacks such as high cost of an ultracentrifuge and rotors, time-consuming and labor-intensive protocol, low yield considering initial biofluid volume and low throughput, development of new EV isolation approaches is still ongoing. Here we compare three methods for isolating the most studied EV subtype, small extracellular vesicles (sEVs), from human plasma: ultracentrifugation (UC), express asymmetric depth filtration (ExADFi), and anti-CD9 immunoaffinity capture (AS-CD9) with focus on their Raman and proteomic profiles. For all three methods, purity and quality of the sEV isolation were assessed based on the level of contamination of the sEV fraction with major plasma proteins such as albumin and apolipoproteins (APOA1, APOH, APOA4, APOC2, APOC1, and APOC4). UC showed the highest ratio of protein to nanoparticle concentration. AS-CD9 and ExADFi provided comparable to UC purity and levels of non-vesicular contaminants with AS-CD9 requiring minimal time and labor. ExADFi showed characteristics including purity of the sEV samples, yield, and isolation time that is between the UC and AS-CD9 methods. Raman spectroscopy provided more details about characteristics of the isolated sEVs and confirmed differences observed in the proteomic profiles. The findings demonstrate that the AS-CD9 and ExADFi methods could be appropriate substitutes of the classical UC-based isolation method and be chosen depending on the final requirements and use of the purified sEVs such as further functional and biomarker studies. Show less

Xinjiang Brown cattle are an important beef breed in Northwest China. Although multigenerational selective breeding has improved their growth performance, the accompanying molecular adaptations and potential physiological trade- ofs remain insufficiently elucidated at the systemic level. This study aimed to decipher the dynamic serum proteomic profiles shaped by both ontogeny and generational selection in Xinjiang Brown cattle, and to identify the associated key proteins and pathways. Serum samples from 18 bulls across three genera- tions (A, B, C) at 3 and 9 months of age were analyzed using Tandem Mass Tag (TMT)-based quantitative proteomics. Under stringent quality control (FDR < 1%), 583 high-confidence proteins were identified. Diferentially expressed proteins (DEPs) were screened using thresholds of |fold change| ≥ 1.2 and This study reveals that the breeding strategy for Xinjiang Brown cattle prioritizes shaping a proteomic landscape that promotes growth and metabolism, potentially at the cost of atten- uated immune-vascular reactivity. The identified panel of candidate proteins pro- vides a molecular framework for evaluating breeding outcomes and designing balanced selection strategies. Follow-up research should further investigate the functions of these candidate proteins and validate their predictive value for health and production performance in independent herds. Show less

Early recognition of a risk of Alzheimer's disease (AD) remains a global challenge, and blood proteomic markers are of particular interest for wide-scale diagnostic use. Quantitative multiple reaction monitoring (MRM) approach demonstrates good reproducibility in the characteristic changes in the levels of reported candidate biomarkers (CBs) in different cohorts in AD. Following up on our previous study, we performed a joint analysis of 331 blood plasma samples from two different clinical cohorts of participants, comprising a total of 95 samples from patients with AD, 136 samples from patients with mild cognitive impairment (MCI), and 100 samples from controls. The obtained results confirm the significance of 37 CBs. A logistic regression-based algorithm was used to build protein classifiers, and a total of 21 important proteins were selected, 13 of which (ORM1, APOA4, LBP, HP, FN1, BCHE, APOE, PZP, A1BG, TF, SERPINA7, TTR, and F12) formed a universal panel that demonstrated strong classification performance in distinguishing AD patients from controls (ROC-AUC = 0.90) and in separating stable and progressing patients with MCI (ROC-AUC = 0.81). Overall, the analysis confirms the high potential of the MRM method for validating CBs in independent cohorts. Show less

Apolipoprotein A-IV (ApoA-IV) has been implicated in modulating the gut microbiota. However, chronic high-fat diet (HFD) consumption impairs ApoA-IV signaling and disrupts gut microbial balance, contributing to obesity and insulin resistance. This study aimed to investigate the role of ApoA-IV in shaping the gut microbiota and associated metabolic profiles throughout the lifespan of mice exposed to an HFD. Fecal samples were collected from ApoA-IV knockout (KO) and wild-type mice at five time points for microbiota and metabolite profiling using 16S rRNA gene sequencing and gas chromatography-mass spectrometry, respectively. Lifespan was longest in ApoA-IV-KO mice on a normal diet, while the HFD reduced survival across genotypes. Microbiota analysis revealed diet- and age-dependent shifts, including an elevated Firmicutes/Bacteroidota ratio, altered abundance of Show less

Variants linked to the risk of ischemic stroke have been discovered through genome-wide association studies (GWASs). These variations frequently have little consequences that lack apparent biological significance. Hence, these findings demonstrate that exome sequencing can be highly relevant to stroke, even though stroke is a complex phenotype with various diseases and risk factors. In this case-control investigation, we used ARMS genotyping to investigate the distribution of polymorphic variations in genes associated with stroke susceptibility. In addition to examine the novel gene variations associated with ischemic stroke we utilized the Illumina NovaSeq 6000 platform for whole-exome sequencing (WES). Results identified 11 novel gene variants in the GSTT4 gene by targeted whole-exome sequencing, including one deletion GSTT4p.Asn232LysfsTer6, one insertion c.688₆₈₉insCG, and 9 SNVs c.699 T > C, c.701C > G, c.708G > T, c.710 T > G, c.712A > G, c.712A > G, c.718A > T, c.719G > A, c.721A > T, c.722G > T in the ischemic stroke patients. We also identified several rare, intermediate, and most common gene variants in cholesterol associated genes LDLR, LDLRAD2, LDLRAD3, APOA2, APOA3, APOA4, APOA5, and PCSK9. Also, several common gene variants were reported in MTHFR, KLF14, eNOS3, and ACE by whole-exome sequencing. Furthermore, the eNOS3-GG and eNOS3-GT genotypes were associated with susceptibility to ischemic stroke (OR = 1.95, This case-control study identified 11 novel GSTT4 variants and several known polymorphisms associated with ischemic stroke risk in Saudi patients. These findings highlight population-specific genetic factors that warrant further functional and large-scale validation. Show less

This study evaluated the protective effects of naringin (NG) against intestinal injury in 7-day-old piglets infected with porcine epidemic diarrhea virus (PEDV). Eighteen piglets (Duroc × Landrace × Large, body weight = 2.58 ± 0.05 kg) were divided into three treatment groups based on similar body weights and equal numbers of males and females: the blank control group (CON group), the PEDV infection group (PEDV group), and the NG intervention + PEDV infection group (NG + PEDV group) ( Show less

Obesity-induced metabolic inflammation is a key driver of chronic kidney disease (CKD), with immune dysregulation, particularly among lymphocytes, contributing to early disease pathology. To explore the role of apolipoprotein A4 (Apoa4) in regulating immune cell metabolism and function, we establish high-fat diet-induced obese (DIO) models using wild-type and Show less

Insulin resistance (IR) contributes to atherogenic dyslipidemia and elevated ASCVD risk. Apolipoprotein A1 (ApoA1)-associated lipoproteins have diverse anti-atherogenic functions, but it is unclear whether IR drives adverse changes in their proteomic composition. We hypothesized that IR is associated with an atherogenic ApoA1 proteome and that insulin-sensitizing interventions would improve its composition. We studied 861 participants without diabetes (age 47 ± 12 years, 65.5% female). IR was directly measured using the steady-state plasma glucose (SSPG) concentration via the insulin suppression test. ApoA1-associated proteins were quantified by mass spectrometry. A subset underwent interventions for 3 months (N total 108): pioglitazone, PIO Show less