ABSTRACTBackgroundChronic spontaneous urticaria (CSU) is a mast cell‐driven disease associated with systemic inflammation and altered immune responses. Haptoglobin (HP), an acute‐phase glycoprotein, exhibits antioxidative and immunomodulatory properties, while zonulin, the precursor of HP‐2, regulates epithelial barrier integrity. We investigated the clinical relevance of serum HP and zonulin in CSU and their association with treatment outcomes.MethodsSerum HP and zonulin levels were measured by ELISA in 124 CSU patients and 57 healthy controls (HCs). In 62 CSU patients, follow‐up samples were obtained after 3 months of treatment. Clinical outcomes included the urticaria activity score over 7 days (UAS7) and the urticaria control test (UCT).ResultsSerum HP levels were significantly higher in CSU patients than in HCs (median 1145.1 vs. 839.2 μg/mL, p < 0.001), whereas zonulin levels did not differ. HP correlated positively, but weakly, with the white blood cell count, C3 and C‐reactive protein (all rho ≈ 0.2), and negatively with disease duration and eosinophil percentage. Zonulin correlated negatively with UCT scores but not with HP. After treatment, HP decreased significantly (p < 0.001), with greater reductions in patients showing a ≥ 12‐point improvement on the UAS7. Baseline HP was higher in patients who achieved complete control (UCT = 16) than others (p = 0.017). ROC analysis identified baseline HP ≥ 1249 μg/mL as an optimal cutoff, confirmed as an independent predictor of complete control (odds ratio = 4.23, p = 0.029).ConclusionSerum HP is elevated in CSU patients, decreases with treatment, and independently predicts complete urticaria control. HP may serve as a potentially prognostic biomarker for CSU.

Haptoglobin (HP) 2-2 phenotype has been suggested as a risk factor for cardiovascular disease (CVD) and to modulate fenofibrate benefit on CVD risk in Type 2 diabetes. However, little is known as to whether HP levels modulate CVD risk and fenofibrate response. Haptoglobin phenotype and levels were determined in 8047 Fenofibrate Intervention and Event Lowering in Diabetes trial participants at baseline and randomization (after a 16 week run-in period, including a 6 week fenofibrate therapy) and their association with new on-trial total CVD events over 5 years was assessed. Higher baseline HP levels were associated with total CVD events in the placebo group {n = 4030, hazard ratio [95% confidence interval (CI)] = 1.30 [1.02-1.66] for HP level Tertile 3 vs. Tertile 1, P = 0.035}. This was driven by participants with HP 1-1 phenotype (P for interaction = 0.011). Participants with the lowest baseline HP level tertile and HP 1-1 phenotype tended to have the lowest CVD risk, whereas CVD risk was similar in other participants, regardless of HP phenotype and levels. Fenofibrate benefit on total CVD events did not differ significantly by HP phenotypes, baseline HP level tertiles, or tertiles of change in HP levels by fenofibrate during active run-in. Higher baseline HP levels were associated with a higher CVD risk, especially in participants with HP 1-1 phenotype. A lower CVD risk is found only in Type 2 diabetes participants with both the HP 1-1 phenotype and low baseline HP levels. Fenofibrate benefits on CVD risk reduction did not differ by HP levels or phenotype. We evaluated the relationship of different forms or phenotypes of the blood protein, haptoglobin, and its circulating levels with the risk of developing CVD and fenofibrate benefit in patients with Type 2 diabetes, and showed that both HP phenotype and levels are important to assess CVD risk in patients with Type 2 diabetes, although fenofibrate did not modulate CVD risk via HP phenotype or level.Higher baseline HP levels were associated with higher CVD risk, especially in participants with HP 1-1 phenotype.Fenofibrate benefit on CVD did not differ by HP phenotype and baseline or change in HP levels.

Peripheral innate immune signature links migraine and depression: Identification of PTX3 and HP as shared diagnostic biomarkers.

BackgroundA structural variant (SV) of haptoglobin (HP) was shown to interact with apolipoprotein E (APOE) genotypes to modify Alzheimer's disease (AD) risk effect. The two‐tandem exon repeat of HP1 to HP2 affects the structure and antioxidative function of the HP protein. HP protein has been found to bind APOE in the brain tissue of human AD patients, and is able to bind amyloid‐beta in vitro. Our previous discovery was conducted in an array‐genotyped European‐descent cohort, and here we replicate this with whole‐genome sequenced non‐Hispanic White (NHW) and African‐American (AFR) datasets.MethodsHP1/HP2 alleles were imputed from 5155 NHW and 4133 AFR samples in the Alzheimer's Disease Sequencing Project using a published panel. We encoded HP status as the number of HP2 alleles. Each allele of APOE was encoded separately as APOE1 and APOE2, and as a single dosage variable ε2‐ε3‐ε4, assigning APOE ε2 baseline risk, with linearly additive risk in ε3 and ε4. Logistic regression was used to model AD associations, HP‐by‐APOE interactions, and a three‐way term for HP‐APOE1‐APOE2. We accounted for deviations from additivity with stratified analysis for APOE carriers of each allele. Age‐of‐onset, sex, and 3 principal components are used as covariates.ResultsThe imputed HP2 SV frequency was 0.38 in NHW and 0.59 in AFR. Although HP did not independently associate with AD, we detected interaction effects between HP2 and APOE on AD risk. Increase in HP2 allele count increased the risk effect of APOE ε4 and protective effect of APOE ε2 (NHW OR=1.43, p = 0.083; AFR OR=1.50, p = 0.029). In analysis stratified by one APOE allele, we find the largest interaction effect between HP and the remaining APOE allele (HP‐APOE‐alt) in APOE ε2 carriers (NHW OR=1.31, p = 0.121; AFR OR=1.43, p = 0.025). Modeled AD risk is lowest in individuals homozygous for both APOE ε2 and HP2.ConclusionWe find that a structural variant of HP modifies the risk effect of APOE alleles. Our findings in an independent dataset corroborate elements of our previous discovery of this interaction effect.

Glycosylation is a post-translational modification process that plays a fundamental role in malignant transformation. Moreover, aberrant glycosylation is known to be associated with cancer progression. Thus, the characterization of cancer-specific protein glycosylation profiles might reveal important diagnostic and/or prognostic biomarkers for cancer. In the present study, we have analysed serum protein and glycoprotein profiles during breast cancer progression using a mouse model. Specifically, 4T1 tumour cells were injected into the mammary fat pad of BALB/c mice to induce tumours. Sera samples were subsequently collected weekly for four weeks and examined using two-dimensional electrophoresis (2D-E) coupled with lectin-based analysis, followed by mass spectrometry. This glycoproteomic profiling identified eight differentially expressed proteins, of which alpha-1 protease inhibitor 2, contraption (CON), haptoglobin (HP), and kininogen-1 were significantly up-regulated during the first 4 weeks of tumour progression. Notably, aberrantly N-glycosylated prothrombin was also detected in sera samples from all mice over the 4 weeks post-tumour injection. Additionally, O-glycosylated alpha-2-macroglobulin, CON, and HP were detected in weeks 1 and 2, whereas O-glycosylated alpha-2-HS-glycoprotein and CON were detected on weeks 3 and 4 post-implantation. Our findings indicate that the combination of 2D-E with lectin-based chromatography represents an effective approach for identifying prognostic biomarkers for breast cancer.

This study aimed to identify hematological markers that can objectively assess postpartum recovery in dairy cows, which is essential for determining an individualized voluntary waiting period (VWP). Twenty Holstein cows were divided into early recovery and late recovery groups based on their clinical uterine recovery. Blood samples were collected from coccygeal vessels and analyzed for inflammatory and metabolic markers, including serum amyloid A (SAA), haptoglobin (HPT), cortisol (COR), substance P (SP), interleukin-6 (IL-6), total cholesterol (T-Chol), beta-hydroxybutyrate, and non-esterified fatty acids (NEFA). Both SAA and HPT significantly decreased after recovery compared to immediately after parturition (p < 0.001), whereas T-Chol and NEFA significantly increased and decreased over time, respectively, indicating recovery from a negative energy balance status. No significant changes were observed in COR, SP, or IL-6 levels. Thus, SAA and HPT are sensitive indicators of postpartum inflammation, whereas T-Chol and NEFA levels reflect metabolic recovery. Using these hematological markers enables a more objective and customized setting of VWP for each cow, potentially improving reproductive efficiency and farm profitability. Further studies are warranted to establish cutoff values for each parity and validate the applicability of individualized VWP models.

Selected comparative aspects and unexpected findings in acute phase proteins and other biomarkers of animal health and welfare.

BackgroundMyocardial infarction (MI) is one of the leading causes of death worldwide. Finding reliable diagnostic biomarkers and gaining a deeper understanding of their role in the immune microenvironment is of great significance for improving clinical prognosis.MethodThis study integrated multiple datasets from GEO (GSE141512, GSE95368, GSE269269) and TCGA data, and used various bioinformatics methods such as weighted gene co-expression network analysis (WGCNA), immune cell infiltration analysis, and single-cell RNA sequencing analysis to screen key genes related to the occurrence and development of myocardial infarction. We initially validated the results using a proteomic dataset (GSE95368) and clinical samples analyzed by qPCR. Critically, the dysregulation and diagnostic value of Haptoglobin (HP) were further confirmed in multiple independent external cohorts (GSE66360, and others.), solidifying its reliability as a biomarker.ResultThe study found that Haptoglobin (HP) is a key gene significantly upregulated in myocardial infarction, and it exhibits high diagnostic value (AUC=0.833) in the proteomic dataset (GSE95368). Single-cell sequencing analysis showed that HP is significantly highly expressed in classical monocyte of MI patients, and this finding was validated by qPCR experiments in clinically collected classical monocytes samples (p<0.05). Functional enrichment analysis implicated HP in immune responses and ferroptosis.ConclusionThe HP gene is a potential diagnostic biomarker for myocardial infarction, and its specific high expression in classical monocytes implies a potential role in the pathological process of myocardial infarction by regulating the immune microenvironment. This study provides a new research direction for the diagnosis and immune-targeted therapy of myocardial infarction, and has important clinical translational value.

BackgroundInfantile epileptic spasm syndrome (IESS) presents significant therapeutic challenges, with the molecular mechanisms underlying variable responses to adrenocorticotropic hormone (ACTH) remaining poorly understood. This study aimed to identify ACTH-specific therapeutic biomarkers in IESS patients with effective (EF) and ineffective (IEF) responses to ACTH, providing potential clues for therapeutic interventions and insights into IESS pathogenesis.MethodsSixty IESS patients were recruited and allocated into the EF group (n = 30) and IEF group (n = 30), alongside 40 age- and gender-matched healthy controls. Plasma samples were analyzed using data-independent acquisition (DIA) proteomics to identify differentially expressed proteins (DEPs). Functional annotation of DEPs was conducted using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Receiver operating characteristic (ROC) curve analysis was employed to construct a diagnostic biomarker model. Enzyme-linked immunosorbent assay (ELISA) validation ensured the robustness of our findings.ResultsA total of 114 proteins were identified as uniquely associated with the EF group. GO and KEGG analyses revealed DEPs in pathways related to humoral immune response regulation, phagocytosis, complement and coagulation cascades, and metabolic processes. ROC curve analysis highlighted complement component 8 beta (C8β), Plasminogen (PLG), Haptoglobin (HP), Aldolase A (ALDOA), and Collagen Type XVIII Alpha 1 (COL18A1) as potential predictive biomarkers for ACTH efficacy, each achieving an area under the curve value above 0.8. Quantitative ELISA validation confirmed higher levels of C8β and PLG, and lower levels of HP, ALDOA, and COL18A1, in the EF group compared to the IEF group, consistent with the DIA results.ConclusionsThese findings offer novel insights into the molecular mechanisms underlying ACTH response variability in IESS and propose candidate plasma protein biomarkers for predicting ACTH treatment efficacy. This study, combining DIA-MS proteomics with targeted ELISA validation in plasma from individuals with IESS, provides evidence that the identified proteins warrant further investigation as candidate biomarkers to refine therapeutic strategies and monitor patient responses.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12014-025-09559-z.

Abstract Ruminal acidosis is a major metabolic disorder in agriculturally important ruminants, such as cattle and sheep. A common symptom of ruminal acidosis is systemic inflammation. This study aimed to identify the underlying causes of systemic inflammation during acute ruminal acidosis using both in vivo and in vitro experiments. In the in vivo experiments, 12, 9-month-old sheep were randomly assigned to two groups, 6 sheep per group. Sheep in one group (the acidotic group) were fasted for 24 h and then provided ad libitum access to a 1:1 mix of whole corn and wheat flour to induce acute ruminal acidosis. Sheep in the other group (the control group) were not fasted and had ad libitum access to alfalfa hay. Rumen fluid and blood samples were collected every 12 h, and all sheep were euthanized 48 h after the introduction of the concentrate diet for the liver tissue collection. Data were analyzed using a linear-mixed effects model, with treatment and time point as fixed effects and animal as a random effect. Acute ruminal acidosis in the sheep fed the concentrate diet was confirmed by ruminal pH values below 5.0, marked increases in ruminal concentrations of free lipopolysaccharides (LPS), L- and D-lactic acids, and symptoms such as loss of appetite and diarrhea. Concentrations of major volatile fatty acids (acetic, propionic and butyric acids) in the rumen and plasma were lower in the acidotic sheep compared to the controls (P &amp;lt; 0.05). Plasma concentrations of LPS, L-lactic acid and D-lactic acid were higher in the acidotic sheep (P &amp;lt; 0.05). Hepatic mRNA expression of haptoglobin (HP), serum amyloid a2 (SAA2), interleukin 1 alpha (IL1A) and interleukin 6 (IL6) as well as plasma concentrations of IL1A and IL6 were higher in the acidotic sheep compared to controls (P &amp;lt; 0.05). These differences demonstrated that the acidotic sheep developed systemic inflammation, which was associated with increased ruminal and plasma concentrations of LPS and L- and D-lactic acids. In the in vitro experiments, hepatocytes and peripheral blood mononuclear cells (PBMC) isolated from normal sheep were cultured with or without LPS, L-lactate or D-lactate. In hepatocyte cultures, LPS increased the expression of HP, SAA2, IL1A, IL1B, IL6, and tumor necrosis factor (TNF) mRNAs (P &amp;lt; 0.05), while D-lactate increased the expression of HP and IL6 mRNAs (P &amp;lt; 0.05), and L-lactate decreased the expression of IL6 mRNA (P &amp;lt; 0.05). In PBMC cultures, both LPS and D-lactate increased the expression of HP, SAA2, IL1A, IL1B and IL6 mRNAs (P &amp;lt; 0.05), while L-lactate increased the expression of HP mRNA only (P &amp;lt; 0.05). Collectively, these in vivo and in vitro findings support the conclusion that both increased concentrations of LPS and D-lactic acid contribute to systemic inflammation during acute ruminal acidosis.

ABSTRACTBackground and Aim:Dairy cows in the transition period are highly vulnerable to oxidative stress and immune suppression, which increases the risk of metabolic and infectious diseases. Vitamin E (VE) and selenium (Se) are essential antioxidants known to mitigate these challenges, but their combined effects remain underexplored in transition cows. This study aimed to evaluate the effects of VE and Se supplementation–individually and in combination–on oxidative stress biomarkers, immune function, disease incidence, reproductive performance, and economic outcomes in transition dairy cows.Materials and Methods:Forty Holstein cows with similar baseline characteristics were randomly assigned to four groups (n = 10 each): Control (basal diet), VE (3,000 IU/head injected on days 7 and 14 postpartum), Se (1.5 mg/kg body weight orally from calving), and VE + Se (both interventions). Blood samples were collected on calving day and at 7, 14, and 21 days postpartum. Parameters assessed included non-esterified fatty acid (NEFA), β-hydroxybutyrate (BHB), aspartate aminotransferase (AST), blood urea nitrogen, total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), interleukin (IL)-1β, IL-6, haptoglobin (HP), milk yield, disease incidence, and economic performance. Receiver operating characteristic (ROC) curves assessed VE and Se’s predictive value for disease.Results:Combined VE + Se supplementation significantly increased plasma VE and Se levels and improved antioxidant capacity (↑T-AOC, SOD, GSH-Px; ↓MDA) and immune markers (↓IL-1β, IL-6, HP). NEFA and BHB were reduced without affecting AST. The VE + Se group showed significantly lower incidences of mastitis, metritis, and ketosis (p < 0.05). ROC analysis demonstrated high predictive value of plasma VE and Se for disease risk (area under the curve up to 0.973). Economic analysis showed the highest net profit (¥111.91/day) in the VE + Se group.Conclusion:Combined VE and Se supplementation during the transition period enhances antioxidant and immune function, reduces metabolic disease incidence, and improves productivity and profitability in dairy cows. These findings support integrated micronutrient strategies for periparturient health management. Larger-scale and long-term studies are recommended to confirm these outcomes and explore underlying mechanisms.

Introduction and Objective: Haptoglobin (HP) is a circulating glycoprotein with antioxidant and anti-inflammatory properties. It scavenges toxic free hemoglobin. Cardiovascular disease (CVD) risk in Type 2 diabetes (T2D) patients may differ by HP phenotype. It is unknown if CVD risk and fenofibrate benefits differ by HP levels. Methods: HP phenotype and levels were measured in baseline plasma from 8047 T2D adults in the Fenofibrate Intervention and Event Lowering in Diabetes (FIELD) trial by ELISA and immunoturbidimetric assay, respectively. Results: Frequencies of HP 1-1, 2-1 and 2-2 phenotypes were 15.6%, 47.6% and 36.9% respectively. Higher HP level tertiles, but not HP phenotype, were associated with a higher risk of total CVD events, especially coronary heart disease events (P=0.019) and CVD mortality (P=0.003) in placebo group, after adjustment for CVD risk factors (overall P for trend =0.032, Fig. 1A). The HP level association was independently significant only in the HP 1-1 phenotype (P for level/phenotype interaction =0.039, Fig. 1B). Participants with both HP 1-1 phenotype and the lowest HP tertile levels tended to have the lowest CVD risk (Fig. 1C). Fenofibrate benefits on total CVD events did not differ by HP phenotype or levels. Conclusion: HP levels and phenotype help to identify T2D patients at high CVD risk for treatment, especially those with HP 1-1 phenotype. Disclosure K. Ong: None. A.S. Januszewski: None. H. Francis: None. R.L. O'Connell: None. A. Mangani: None. L. Li: None. P.G. Colman: None. D. Sullivan: Other Relationship; Amgen Inc. Research Support; Arrowhead Pharmaceuticals, Inc. Other Relationship; Novartis Pharmaceuticals Corporation, Merck Sharp &amp; Dohme Corp. Research Support; Ionis Pharmaceuticals. J.D. Best: None. R.S. Scott: None. A. Jenkins: Advisory Panel; Abbott. Speaker's Bureau; GlaxoSmithKline plc. Research Support; Abbott, Metronics, Ypsomed AG, Insulet Corporation, Jaeb Center for Health Research, Endogenex, Hemsley Charitable Trust. Speaker's Bureau; CSL Seqirus. Research Support; AbbVie Inc, National Institutes of Health, Juvenile Diabetes Research Foundation (JDRF). A.C. Keech: Research Support; Abbott, Amgen Inc, ASPEN Australia, Mylan. Speaker's Bureau; Novartis AG, Pfizer Inc. Research Support; Kowa Company, Ltd. Speaker's Bureau; Sanofi. Research Support; AbbVie Inc, Viatris Inc. Funding Laboratoires Fournier, Dijon, France and the National Health and Medical Research Council of Australia (457103, 1024105, 1037786, 1105467, 1121272, 1137071 and 2018537)

Protein electrophoresis and acute phase proteins are valuable clinical diagnostic tools to identify inflammation. This study evaluated capillary zone electrophoresis (CZE) for clinically normal giant pandas (Ailuropoda melanoleuca) against individuals that were clinically ill. Assay validation for C-reactive protein (CRP), serum amyloid A (SAA), and haptoglobin (HP) was also attempted. Using CZE, total protein, alpha-1 and -2, beta globulins, beta-2 globulins, and gamma globulins were observed to be significantly higher, and albumin:globulin, pre-albumin, and albumin were significantly lower in clinically abnormal giant pandas (p < 0.05). Among the tested APPs, reactivity was only found for CRP reagents and was significantly higher in abnormal giant panda (p = 0.045).

Colorectal cancer (CRC) is a widespread and serious global malignancy. This study aimed to investigate the clinical relevance of galanin (GAL) and haptoglobin (HP) as new diagnostic CRC biomarkers. An enzyme-linked immunosorbent assay was used to determine the GAL and HP levels in the serum of 88 patient with CRC and 88 healthy controls. The carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) contents were quantitatively evaluated via electrochemiluminescence assay. Receiver operating characteristic (ROC) curves were created to identify the diagnostic importance of single and combined CRC detection. The patients' relevant diagnostic and treatment data were collected from their medical records. Statistical analysis methods were employed to examine the relationship between these indicators and the clinical pathological parameters. Patients with CRC exhibited significantly decreased and increased serum GAL and HP levels, respectively, compared with the healthy controls (P < 0.0001 for both). Furthermore, the HP level was positively correlated with tumor T stage (P = 0.0124). The area under the curve (AUC) of the ROC values for GAL and HP was 0.744 and 0.712, respectively, indicating their diagnostic efficiency. The combination of GAL and HP increased the AUC to 0.753, and when both were integrated with CEA and CA19-9, the AUC further increased to 0.893. This study shows that serum GAL and HP can be used as potential noninvasive diagnostic biomarkers for CRC.

Haptoglobin (HP) phenotype has been reported to modulate fenofibrate benefit on coronary artery disease in type 2 diabetes. It is unknown whether HP phenotype and levels modulate fenofibrate benefit on sight-threatening diabetic retinopathy (STDR). In plasma from 8,047 Australasian adults with type 2 diabetes in the FIELD trial, HP phenotype was determined, and HP levels were measured at baseline and after six-week fenofibrate run-in. There were 307 new first on-trial STDR events over five years. Baseline HP levels and phenotype were not related to STDR risk. Fenofibrate benefit on STDR vs. placebo (-32% overall), was greatest in participants with the lowest baseline HP level tertile (hazard ratio [95% CI] 0.41 [0.26-0.65], vs. 0.82 [0.56-1.21] and 0.84 [0.56-1.27] for tertiles 2 and 3 respectively, P for heterogeneity=0.019). During run-in, fenofibrate reduced HP levels by 20.7%. However, fenofibrate benefit on STDR did not differ significantly by HP phenotype or change in HP levels during run-in after adjustment for confounding factors. Regarding STDR, fenofibrate benefit is greatest in type 2 diabetes patients with the lowest baseline HP levels, which may reflect patients more susceptible to oxidative retinal injury. All HP phenotypes benefit from fenofibrate.

Fatty liver is a major metabolic disorder in perinatal dairy cows, characterized by elevated plasma concentrations of nonesterified fatty acids (NEFA) and hepatic inflammation. Glutathione S-transferase mu 2 (GSTM2), a phase II detoxification enzyme, regulates cellular antioxidant and detoxification processes in nonruminants. However, its involvement in NEFA-induced hepatic inflammation in dairy cows with fatty liver remains unclear. This study aimed to elucidate the role of GSTM2 in mediating hepatic inflammation caused by elevated NEFA levels in dairy cows with severe fatty liver. An in vivo study was conducted using 10 healthy cows (hepatic triacylglycerol [TG] content <1%) and 10 cows with severe fatty liver (hepatic TG content >10%), matched for the number of lactations (median = 3, range = 2-4) and DIM (median = 9 d, range = 3-15 d). Liver tissue and blood samples were collected before feeding. Compared with healthy cows, cows with severe fatty liver had higher plasma concentrations of NEFA, BHB, haptoglobin (HP), plasma amyloid A (SAA), and lower plasma concentration of glucose. These cows also showed significantly lower abundance of hepatic GSTM2 and overactivated hepatic inflammatory pathways, as indicated by increased abundance of phosphorylated inhibitor of κB (IκB)α and nuclear factor κB (NF-κB) p65, NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing CARD (ASC), and caspase-1 (CASP1), as well as mRNA levels of tumor necrosis factor α (TNFA), IL6, and IL1B. In vitro, hepatocytes isolated from 5 healthy calves (1 d old, fasted female, 30-40 kg of BW) were used to determine the effects of GSTM2 on hepatic inflammation. First, hepatocytes were treated with NEFA (1.2 mM) for varying durations (0.5, 1, 3, 6, 9, 12, 15, or 18 h). The NEFA treatment significantly increased the phosphorylation of IκBα and NF-κB p65, protein abundance of NLRP3, ASC and CASP1, and mRNA levels of TNFA, IL6 and IL1B, peaking at 9 and 12 h. Second, hepatocytes were treated with different concentrations of NEFA (0, 0.6, 1.2, or 2.4 mM) for 9 h, which decreased GSTM2 protein and mRNA abundance. Meanwhile, GSTM2 was silenced using small interfering RNA or overexpressed using adenovirus for 48 h in hepatocytes, followed by NEFA treatment. Silencing GSTM2 augmented the NEFA-induced increase in phosphorylation of IκBα and NF-κB p65, as well as protein abundance of NLRP3, ASC and CASP1, and mRNA levels of TNFA, IL6 and IL1B. Conversely, overexpression of GSTM2 mitigated these inflammatory signals upon NEFA treatment. In summary, these findings indicate that GSTM2 plays a crucial role in modulating NEFA-induced hepatic inflammation. Targeting GSTM2 may offer new strategies to treat or prevent fatty liver disease in dairy cows.

Schizophrenia (SZ), a chronic and widespread brain disorder, presents with complex etiology and pathogenesis that remain inadequately understood. Despite the absence of a universally recognized endophenotype, peripheral blood mononuclear cells (PBMCs) serve as a robust model for investigating intracellular alterations linked to SZ. To preliminarily investigate potential pathogenic mechanisms and identify novel biomarkers for SZ. PBMCs from SZ patients were subjected to integrative transcriptomic and proteomic analyses to uncover differentially expressed genes (DEGs) and differentially expressed proteins while mapping putative disease-associated signaling pathways. Key findings were validated using western blot (WB) and real-time fluorescence quantitative PCR (RT-qPCR). RNAi-lentivirus was employed to transfect rat hippocampal CA1 neurons in vitro, with subsequent verification of target gene expression via RT-qPCR. The levels of neuronal conduction proteins, including calmodulin-dependent protein kinase II (caMKII), CREB, and BDNF, were assessed through WB. Apoptosis was quantified by flow cytometry, while cell proliferation and viability were evaluated using the Cell Counting Kit-8 assay. The integration of transcriptomic and proteomic analyses identified 6079 co-expressed genes, among which 25 DEGs were significantly altered between the SZ group and healthy controls. Notably, haptoglobin (HP), lactotransferrin (LTF), and SERPING1 exhibited marked upregulation. KEGG pathway enrichment analysis implicated neuroactive ligand-receptor interaction pathways in disease pathogenesis. Clinical sample validation demonstrated elevated protein and mRNA levels of HP, LTF, and SERPING1 in the SZ group compared to controls. WB analysis of all clinical samples further corroborated the significant upregulation of SERPING1. In hippocampal CA1 neurons transfected with lentivirus, reduced SERPING1 expression was accompanied by increased levels of CaMKII, CREB, and BDNF, enhanced cell viability, and reduced apoptosis. SERPING1 may suppress neural cell proliferation in SZ patients via modulation of the CaMKII-CREB-BDNF signaling pathway.

Obstructive sleep apnea (OSA) patients have varying degrees of cognitive impairment, but the specific pathogenic mechanism is still unclear. Meanwhile, poor compliance with continuous positive airway pressure (CPAP) in OSA prompts better solutions. This study aimed to identify differentially expressed genes between the non-obese OSA patients and healthy controls, and to explore potential biomarkers associated with cognitive impairment. Cohorts of healthy control (n = 20) and non-obese, treatment-naïve OSA patients (n = 20) were recruited. We collected their peripheral blood mononuclear cells and neutrophils, and their cognitive performances were evaluated by the Montreal Cognitive Assessment (MoCA). The differentially expressed genes were identified by bioinformatic analysis and confirmed by PCR. Imbalanced immune cell proportions were assessed by Cibersort. Biomarkers related to enriched cellular pathways were measured by ELISA. OSA patients showed a significant decline in overall cognitive function and were associated with higher daytime sleepiness scores. Multiple signaling pathways were enriched in the non-obese OSA cohort, including upregulation of neutrophil-degranulation. Increased monocyte proportion and decreased NK cell proportion were figured out. The relevant genes, including upregulated defensin alpha 4 (DEFA4), haptoglobin (HP), survivin (BIRC5), and suppressed interferon gamma (IFNG) expression were detected. The relative expression of DEFA4 was significantly correlated with the MoCA score and sleep parameters. Biomarkers such as myeloperoxidase (MPO), H2O2, and lipocalin-2, as representatives of neutrophils’ activation, elevated significantly in the OSA group. The data demonstrated a positive correlation between MPO and oxygen desaturation index (ODI) and a negative correlation between MPO and lowest oxygen saturation (LSaO2). The level of Lipocalin-2 was positively correlated with apnea-hypopnea index (AHI) and ODI and negatively correlated with LSaO2 and MoCA score. We also observed a negative correlation between H2O2 and mean oxygen saturation (MSaO2). Degranulation of neutrophils was activated in non-obese OSA patients without other complications. The process is related to OSA severity and cognitive impairment, implying its role in pathogenesis.

Acute ruminal acidosis is a major metabolic disorder in agriculturally important ruminants, such as cattle and sheep. A common symptom of acute ruminal acidosis is systemic inflammation. This study aimed to identify the underlying causes of systemic inflammation during acute ruminal acidosis using both in vivo and in vitro experiments. In the in vivo experiments, twelve 9-month-old sheep were randomly assigned to two groups, six sheep per group. Sheep in the acidotic group were fasted for 24 h and then provided ad libitum access to a 1:1 mix of whole corn and wheat flour to induce acute ruminal acidosis. Sheep in the control group were not fasted and had ad libitum access to alfalfa hay. Rumen fluid and blood samples were collected every 12 h, and all sheep were euthanized 48 h after the introduction of the concentrate diet. Acute ruminal acidosis in the sheep fed the concentrate diet was confirmed by ruminal pH < 5.0, elevated levels of free lipopolysaccharides (LPS), L- and D-lactic acids, and symptoms like loss of appetite and diarrhea. Concentrations of acetic, propionic, and butyric acids in rumen fluid and plasma were lower in the acidotic sheep (P < 0.05). Plasma concentrations of LPS, L-lactic acid and D-lactic acid were greater in the acidotic sheep (P < 0.05). Hepatic mRNA expression of haptoglobin (HP), serum amyloid a2 (SAA2), interleukin 1 alpha (IL1A) and interleukin 6 (IL6) as well as plasma concentrations of IL1A and IL6 were greater in the acidotic sheep compared to controls (P < 0.05). These changes indicated systemic inflammation, which was associated with increased rumen fluid and plasma concentrations of LPS and L- and D-lactic acids. In the in vitro experiments, peripheral blood mononuclear cells (PBMC) and hepatocytes isolated from normal sheep were cultured with or without LPS, L-lactate or D-lactate. In PBMC cultures, both LPS and D-lactate increased the expression of HP, SAA2, IL1A, IL1B, and IL6 mRNAs (P < 0.05), while L-lactate increased the expression of HP mRNA only (P < 0.05). In hepatocyte cultures, LPS increased the expression of HP, SAA2, IL1A, IL1B, IL6, and tumor necrosis factor (TNF) mRNAs (P < 0.05), while D-lactate increased the expression of HP mRNA (P < 0.05), and L-lactate decreased the expression of IL6 mRNA (P < 0.05). Collectively, these in vivo and in vitro findings support the conclusion that both increased concentrations of LPS and D-lactic acid contribute to systemic inflammation during acute ruminal acidosis.

COVID-19 is a highly contagious viral disease that primarily affects the respiratory system and occasionally the gastrointestinal system, and it was declared a pandemic in 2020. Haptoglobin is an acute-phase protein and a potent antioxidant in the body, which exerts its antioxidant effect by binding to free hemoglobin. Haptoglobin has three main variants (Hp1-1, Hp1-2, Hp2-2), each with different antioxidant capacities. The purpose of this study is to investigate frequency of the haptoglobin variants in COVID-19 patients compared to a control group. This study was conducted on 148 COVID-19 patients and 145 healthy individuals from the Sistan and Baluchestan province. DNA was isolated from whole blood using the salt precipitation method, and the determination of haptoglobin genotypes (Hp1-1, Hp1-2, and Hp2-2) was performed using Conventional PCR. This study analyzed haptoglobin (HP) genotypes in COVID-19 patients and controls, finding no significant difference in HP variant frequencies between groups (p= 0.529). However, the HP1-2 genotype was associated with a twofold increased COVID-19 risk in men (OR=2.069, p= 0.021), and the HP1 allele significantly raised infection risk (OR= 1.62, p= 0.039). Hospitalizations and respiratory symptoms were significantly higher in COVID-19 patients (p= 0.0001 and p= 0.0176, respectively). These results suggest that haptoglobin variants are not risk factors for COVID-19 infection in the overall population (both males and females). However, men with the HP1-2 genotype are 1.9 times more likely to develop COVID-19 infection compared to men with HP1-1 and HP 2-2 genotypes.