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Simple Summary: The diagnosis of nasal cavity disease (ND) in dogs typically requires comprehensive diagnostics under general anesthesia including cross-sectional imaging and rhinoscopy. Therefore, dogs with nasal discharge are often only presented for diagnostics after weeks of symptomatic or even antibiotic therapy, even though primary bacterial rhinitis is uncommon. In a previous study, the platelet-to-lymphocyte ratio (PLR) differed significantly between dogs with malignant tumors and benign pathology. To further investigate the value of blood tests for diagnosis, the aim of this second part of the study was to evaluate serum markers for their diagnostic utility and whether they may be helpful in making the diagnosis of ND quickly and cost-effectively. Dogs with additional diseases detected by a whole-body CT scan and blood tests were excluded. No significant differences were found between groups for the here evaluated markers C-reactive protein, haptoglobin, and 25-hydroxyvitamin-D. In this prospective blinded study, canine C-reactive protein (c-CRP), haptoglobin (HPT), and 25-hydroxyvitamin-D (25(OH)D) were investigated for their diagnostic value in 55 dogs with nasal cavity disease (ND). After comprehensive diagnostics including a culture-dependent microbiological examination (ME) of nasal swabs, 17 dogs were excluded due to additionally detected systemic diseases or steroid pre-treatment. Included were 25 dogs with malignant ND (13 carcinomas and 12 sarcomas) and 30 dogs with benign ND (7 benign tumors, 13 idiopathic rhinitis (IR), and 10 others), as well as 10 controls. In none of the 72 dogs with ND was primary bacterial rhinitis diagnosed. Although within the reference interval, compared to the controls, c-CRP was significantly higher in dogs with ND in general and in every subgroup except for benign tumors. Serum HPT concentrations were not different among groups. Compared to the controls, 25(OH)D concentrations were significantly lower (p = 0.041) in malignant ND and sarcomas (p = 0.025). Despite pre-treatment with antibiotics (40/54; 74.1%), in 23/51 (45%) dogs, the ME was positive. Cultivated bacteria did not differ significantly between nasal diseases. The serum markers were not significantly different regarding the positivity of ME. In conclusion, the investigated serum markers were not clinically useful for the reliable detection of canine ND, as was the ME. Because of the low number of dogs with IR and positive or negative ME, further studies regarding c-CRP are needed in a larger group of IR dogs without concomitant diseases to reliably evaluate its utility in IR dogs with suspected secondary bacterial nasal infection. [ABSTRACT FROM AUTHOR]

Simple Summary: Metritis, a condition affecting up to 20% of cows after giving birth, negatively impacts animal welfare and dairy farm profitability by reducing productivity and reproduction. This study compared the biochemical profiles of healthy cows and those with metritis, analyzing 25 salivary and 31 serum analytes alongside various health parameters. Key findings showed that cows with metritis had elevated levels of certain biomarkers related to stress, inflammation, and metabolism in both saliva and serum. Specifically, eight salivary biomarkers including adenosine deaminase (ADA) and haptoglobin (Hp) and eight serum biomarkers including ADA, Hp, and serum amyloid A (SAA) were significantly higher in affected cows. In contrast, six biomarkers including total esterase (TEA) and albumin were lower in the serum. This study highlights the potential of using saliva as a non-invasive source for identifying biomarkers in cows with metritis, offering a new approach to diagnose and manage this condition. Metritis affects 5–20% of cows after parturition, negatively impacting animal welfare and the profitability of dairy farms, increasing culling rates and costs, and decreasing productivity and reproduction rates. This study compared the results of a comprehensive biochemical panel consisting of 25 salivary and 31 serum analytes between healthy cows (n = 16) and cows with metritis (n = 12). Descriptive parameters such as depression, rectal temperature, body condition score (BCS), heart rate, respiratory rate, mucous color, ruminal motility, vaginal discharge, milk production, and complete hematology analyses were also assessed for comparative purposes. The biochemistry analytes comprised five analytes related to stress, five to inflammation, five to oxidative status, and nineteen to general metabolism. The two-way ANOVA analysis revealed that, in saliva, eight biomarkers (lipase, adenosine deaminase (ADA), haptoglobin (Hp), total proteins, g-glutamyl transferase (gGT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and creatine kinase (CK)) were significant higher in cows with metritis. In serum, eight biomarkers (ADA, Hp, serum amyloid A (SAA), fibrinogen, ferritin, AOPPs/albumin ratio, non-esterified fatty acids (NEFAs), and bilirubin) were significantly higher in cows with metritis, whereas six (total esterase (TEA), albumin, urea, lactate, phosphorus, and calcium) were lower. Of the total number of 23 biomarkers that were measured in both saliva and serum, significant positive correlations between the two biofluids were found for six of them (Hp, FRAP, CUPRAC, AOPPs, urea, and phosphorus). Urea showed an R = 0.7, and the correlations of the other analytes were weak (R < 0.4). In conclusion, cows with metritis exhibited differences in biomarkers of stress, inflammation, cellular immune system, and general metabolism in both salivary and serum biochemistry profiles. These changes were of different magnitudes in the two biofluids. In addition, with the exception of ADA and Hp, the analytes that showed changes in the saliva and serum profiles of cows affected by metritis were different. Overall, this report opens a new window for the use of saliva as potential source of biomarkers in cows with metritis. [ABSTRACT FROM AUTHOR]

Preeclampsia (PE) is a hypertensive pregnancy syndrome associated with target organ damage and increased cardiovascular risks, necessitating antihypertensive therapy. However, approximately 40% of patients are nonresponsive to treatment, which results in worse clinical outcomes. This study aimed to compare circulating proteomic profiles and identify differentially expressed proteins among 10 responsive (R-PE), 10 nonresponsive (NR-PE) patients, and 10 healthy pregnant controls (HP). We also explored correlations between these proteins and clinical data. Plasma protein relative quantification was performed using mass spectrometry, followed by bioinformatics analyses with the UniProt database, PatternLab for Proteomics 4.0, and MetaboAnalyst software (version 6.0). Considering a fold change of 1.5, four proteins were differentially expressed between NR-PE and R-PE: one upregulated (fibronectin) and three downregulated (pregnancy-specific beta-1-glycoprotein 1, complement C4B, and complement C4A). Between NR-PE and HP, six proteins were differentially expressed: two upregulated (clusterin and plasmin heavy chain A) and four downregulated (apolipoprotein L1, heparin cofactor II, complement C4B, and haptoglobin-related protein). Three proteins were differentially expressed between R-PE and HP: one downregulated (transthyretin) and two upregulated (apolipoprotein C1 and hemoglobin subunit beta). These findings suggest a complex interplay of these proteins involved in inflammatory, immune, and metabolic processes with antihypertensive therapy responsiveness and PE pathophysiology. [ABSTRACT FROM AUTHOR]

The association between high-density lipoprotein (HDL) cholesterol and breast cancer (BC) remains controversial due to the high complexity of the HDL particle and its functionality. The HDL proteome was determined in newly diagnosed BC classified according to the molecular type [luminal A or B (LA or LB), HER2, and triple-negative (TN)] and clinical stage of the disease. Women (n = 141) aged between 18 and 80 years with BC, treatment-naïve, and healthy women [n = 103; control group (CT)], matched by age and body mass index, were included. Data-independent acquisition (DIA) proteomics was performed in isolated HDL (D = 1.063–1.21 g/mL). Results: Paraoxonase1, carnosine dipeptidase1, immunoglobulin mMu heavy chain constant region (IGHM), apoA-4, and transthyretin were reduced, and serum amyloid A2 and tetranectin were higher in BC compared to CT. In TNBC, apoA-1, apoA-2, apoC-2, and apoC-4 were reduced compared to LA, LB, and HER2, and apoA-4 compared to LA and HER2. ComplementC3, lambda immunoglobulin2/3, serpin3, IGHM, complement9, alpha2 lysine rich-glycoprotein1, and complement4B were higher in TNBC in comparison to all other types; complement factor B and vitamin D-binding protein were in contrast to LA and HER2, and plasminogen compared to LA and LB. In grouped stages III + IV, tetranectin and alpha2-macroglobulin were reduced, and haptoglobin-related protein; lecithin cholesterol acyltransferase, serum amyloid A1, and IGHM were increased compared to stages I + II. Conclusions: A differential proteomic profile of HDL in BC based on tumor molecular classification and the clinical stage of the disease may contribute to a better understanding of the association of HDL with BC pathophysiology, treatment, and outcomes. [ABSTRACT FROM AUTHOR]

One of the most serious diseases affecting dairy cattle, causing significant losses both in breeding and economy, is mastitis, an inflammation of the mammary gland. Due to the economic importance of this issue, many research teams are striving to develop an easy-to-apply and, most importantly, effective method to prevent mastitis. The use of traditional methods for mastitis detecting and treating, as well as improvement in hygienic conditions, have not yielded the expected results in combating this disease combating. Currently, the main task is to find the tools that would allow for the rapid detection of mastitis and the improvement of udder health in cows while maintaining high milk production, which is essential for the profitability of dairy cattle farming. Accurate and rapid diagnostic tools, with the simultaneous capability of identifying pathogens, may help to reduce losses. Sufficient sensitivity and specificity for tests are required to minimize the number of false-positive and false-negative cases. Efforts are also being made to determine the optimal threshold value for detecting the disease at its earliest possible stage. The estimation of somatic cell count (SCC) as a phenotypic indicator of mastitis is widely used. A more precise parameter for accurately describing udder health is the differential somatic cell count (DSCC). The well-known California Mastitis Test (CMT) is an inexpensive, easy, and rapid method for mastitis detection useful on farms. The latest diagnostic methods for mastitis utilize tests based on the activity of N-acetyl-β-d-glucosaminidase (NAGase) or lactate dehydrogenase (LDH) as well as the determination of acute phase proteins (APPs) in blood serum and milk (such as haptoglobin, serum amyloid A, fibrinogen, and ceruloplasmin). Research also focuses on the genomic improvement of mastitis resistance in successive generations, and for this purpose, many quantitative trait loci (QTLs) and single nucleotide polymorphisms (SNPs) have been identified. In recent years, immunotherapy has become an increasingly common area of research, including vaccinations, T/B cell immunotherapy, RNA immunotherapy, epigenetic immunotherapy, stem cell therapy, and native secretory factors. An important aspect of the control of mastitis is the implementation of strategies that focus primarily on preventing the disease through appropriate breeding and farm management practices. In the forthcoming years, a significant challenge will be the development of universal diagnostic and therapeutic strategies that can be effectively implemented as alternatives to antibiotic therapy. Future research should prioritize the advancement of preventive and therapeutic techniques, such as immunotherapies, bacteriocins, herbal therapy, and nanoparticle technology. [ABSTRACT FROM AUTHOR]

Wilson disease is a genetic disorder of the liver characterized by excess accumulation of copper, which is found ubiquitously on earth and normally enters the human body in small amounts via the food chain. Many interesting disease details were published on the mechanistic steps, such as the generation of reactive oxygen species (ROS) and cuproptosis causing a copper dependent cell death. In the liver of patients with Wilson disease, also, increased iron deposits were found that may lead to iron-related ferroptosis responsible for phospholipid peroxidation within membranes of subcellular organelles. All topics are covered in this review article, in addition to the diagnostic and therapeutic issues of Wilson disease. Excess Cu2+ primarily leads to the generation of reactive oxygen species (ROS), as evidenced by early experimental studies exemplified with the detection of hydroxyl radical formation using the electron spin resonance (ESR) spin-trapping method. The generation of ROS products follows the principles of the Haber–Weiss reaction and the subsequent Fenton reaction leading to copper-related cuproptosis, and is thereby closely connected with ROS. Copper accumulation in the liver is due to impaired biliary excretion of copper caused by the inheritable malfunctioning or missing ATP7B protein. As a result, disturbed cellular homeostasis of copper prevails within the liver. Released from the liver cells due to limited storage capacity, the toxic copper enters the circulation and arrives at other organs, causing local accumulation and cell injury. This explains why copper injures not only the liver, but also the brain, kidneys, eyes, heart, muscles, and bones, explaining the multifaceted clinical features of Wilson disease. Among these are depression, psychosis, dysarthria, ataxia, writing problems, dysphagia, renal tubular dysfunction, Kayser–Fleischer corneal rings, cardiomyopathy, cardiac arrhythmias, rhabdomyolysis, osteoporosis, osteomalacia, arthritis, and arthralgia. In addition, Coombs-negative hemolytic anemia is a key feature of Wilson disease with undetectable serum haptoglobin. The modified Leipzig Scoring System helps diagnose Wilson disease. Patients with Wilson disease are well-treated first-line with copper chelators like D-penicillamine that facilitate the removal of circulating copper bound to albumin and increase in urinary copper excretion. Early chelation therapy improves prognosis. Liver transplantation is an option viewed as ultima ratio in end-stage liver disease with untreatable complications or acute liver failure. Liver transplantation finally may thus be a life-saving approach and curative treatment of the disease by replacing the hepatic gene mutation. In conclusion, Wilson disease is a multifaceted genetic disease representing a molecular and clinical challenge. [ABSTRACT FROM AUTHOR]

Age-associated deep-subcortical white matter lesions (DSCLs) are an independent risk factor for dementia, displaying high levels of CD68+ microglia. This study aimed to characterize the transcriptomic profile of microglia in DSCLs and surrounding radiologically normal-appearing white matter (NAWM) compared to non-lesional control white matter. CD68+ microglia were isolated from white matter groups (n = 4 cases per group) from the Cognitive Function and Ageing Study neuropathology cohort using immuno-laser capture microdissection. Microarray gene expression profiling, but not RNA-sequencing, was found to be compatible with immuno-LCM-ed post-mortem material in the CFAS cohort and identified significantly differentially expressed genes (DEGs). Functional grouping and pathway analysis were assessed using the Database for Annotation Visualization and Integrated Discovery (DAVID) software, and immunohistochemistry was performed to validate gene expression changes at the protein level. Transcriptomic profiling of microglia in DSCLs compared to non-lesional control white matter identified 181 significant DEGs (93 upregulated and 88 downregulated). Functional clustering analysis in DAVID revealed dysregulation of haptoglobin–haemoglobin binding (Enrichment score 2.5, p = 0.017), confirmed using CD163 immunostaining, suggesting a neuroprotective microglial response to blood–brain barrier dysfunction in DSCLs. In NAWM versus control white matter, microglia exhibited 347 DEGs (209 upregulated, 138 downregulated), with significant dysregulation of protein de-ubiquitination (Enrichment score 5.14, p < 0.001), implying an inability to maintain protein homeostasis in NAWM that may contribute to lesion spread. These findings enhance understanding of microglial transcriptomic changes in ageing white matter pathology, highlighting a neuroprotective adaptation in DSCLs microglia and a potentially lesion-promoting phenotype in NAWM microglia. [ABSTRACT FROM AUTHOR]

In this report, different handling conditions at slaughterhouse were studied to assess changes in salivary biomarkers. For this purpose, finishing pigs were divided into two groups, one in which handling was improved to minimize stress (Group A, n = 24, transported and stabled at the slaughterhouse at low density without mixing with unfamiliar animals throughout the whole process) and another one in which animals had a more stressful handling process (Group B, n = 24, transported and stabled at high density with unfamiliar animals). Saliva samples were taken the day before transport to the slaughterhouse at 8:00 a.m. (B0) and 12:00 a.m. (B4), and the day of slaughter just after unloading animals at the slaughterhouse at approximately 8:00 a.m. (S0) and after 4 h of lairage at approximately 12:00 a.m. (S4). Group B showed significantly higher cortisol, total esterase activity, oxytocin, adenosine deaminase and haptoglobin levels than the Group A at both S0 and S4 sampling times, and higher levels of calprotectin and creatine kinase at S4 sampling time. This report indicates that differences in the way in which the pigs are handled at the slaughterhouse can lead to changes in salivary biomarkers and opens the possibility of the use of biomarker at slaughter to monitor handling conditions. [ABSTRACT FROM AUTHOR]

This study aims at identifying molecular biomarkers differentiating responders and non-responders to treatment with Tumor Necrosis Factor inhibitors (TNFi) among patients with axial spondyloarthritis (axSpA). Whole blood mRNA and plasma proteins were measured in a cohort of biologic-naïve axSpA patients (n = 35), pre and post (14 weeks) TNFi treatment with adalimumab. Differential expression analysis was used to identify the most enriched pathways and in predictive models to distinguish responses to TNFi. A treatment-associated signature suggests a reduction in inflammatory activity. We found transcripts and proteins robustly differentially expressed between baseline and week 14 in responders. C-reactive protein (CRP) and Haptoglobin (HP) proteins showed strong and early decrease in the plasma of axSpA patients, while a cluster of apolipoproteins (APOD, APOA2, APOA1) showed increased expression at week 14. Responders to TNFi treatment present higher levels of markers of innate immunity at baseline, and lower levels of adaptive immunity markers, particularly B-cells. A logistic regression model incorporating ASDAS-CRP, gender, and AFF3, the top differentially expressed gene at baseline, enabled an accurate prediction of response to adalimumab in our cohort (AUC = 0.97). In conclusion, innate and adaptive immune cell type composition at baseline may be a major contributor to response to adalimumab in axSpA patients. A model including clinical and gene expression variables should also be considered. [ABSTRACT FROM AUTHOR]

Introduction: Chronic hepatitis C (CHC) is a clinical and pathological syndrome with various causes and is characterized by varying degrees of hepatocellular necrosis and inflammation. It is a significant cause of liver transplantation and liver-related death worldwide. The hepatic manifestations of CHC are typically characterized by slowly progressing liver fibrosis, which is a non-specific and often disproportionate response to tissue damage. A large majority of HCV patients have extrahepatic manifestations with varying degrees of severity. HCV infection is a risk factor for cardiovascular disease and diabetes mellitus, which increases insulin resistance, oxidative stress, and iron overload and causes chronic systemic inflammation. HCV infection is treated using direct-acting antivirals (DAAs) with cure rates of over 95 percent, minimal side effects, and shorter therapeutic courses. Despite the effective elimination of the virus, it seemed pertinent to understand to what extent HCV clearance eliminates or attenuates all the systemic alterations already induced by the virus during infection and chronicity. Objectives: Our study aimed to determine whether eliminating HCV with DAAs alters the severity of liver disease (liver stiffness and liver fibrosis stage by TE) and the metabolic/cellular profile of patients with CHC. Materials and methods: A group of 329 CHC patients from a Gastroenterology and Hepatology outpatient department were prospectively studied. Of these, 134 were also studied with DAAs. The liver fibrosis stage was evaluated by transient elastography (TE) using a FibroScan® device, and two groups were established for the analysis of liver stiffness (LS): mild and moderate stiffness (fibrosis F1 and F2; F1/2) and severe stiffness (fibrosis and cirrhosis F3 and F4; F3/4). Metabolic/cellular parameters were evaluated before and after antiviral treatment using standard methods: alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl-transpeptidase (γ-GT), haptoglobin (Hp), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides (TG), free iron (Fe), transferrin saturation (TS), total iron binding capacity (TIBC), ferritin (Ft), glycemia, insulin, Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) and platelets count. The results were statistically analyzed using SPSS 24.0 for Windows. Results: Comparing the fibrosis stage before and after DAAs treatment, we verify a reduction in LS in 85.7% of patients and an improvement in liver fibrosis stage in 22.2% of them after DAAs treatment. Before DAAs treatment, patients showed a 2.410 risk for higher fibrosis stages (F3/4). Comparing metabolic/cellular parameters before and after DAAs treatment, patients showed lower ALP, AST, ALT, γGT, TG, Fe, TIBC, and Ft values and higher TC, LDL, and Hp values after treatment. As such, HCV elimination reduces iron overload and insulin resistance. On the other hand, it caused dyslipidemia, raising total cholesterol and LDL to levels outside the reference values. The improvement in the liver fibrosis stage by TE was mainly associated with higher baseline platelet count and HDL values and lower insulin resistance. Conclusions: With this study, we were able to contribute to the knowledge of the effects of HCV elimination with DAAs on liver disease and metabolic profile to improve the quality of treatment and follow-up of these patients after HCV elimination. [ABSTRACT FROM AUTHOR]

The majority of patients with Diffuse Large B-cell Lymphoma (DLBCL) will respond to first-line treatment and be cured. However, the disease is heterogeneous, and biomarkers able to discriminate patients with suboptimal prognosis are needed. M2 CD163-positive tumor-associated macrophages (TAMs) were shown to be implicated in DLBCL disease activity regulation. Serum-soluble CD163 (sCD163) functions as a scavenger receptor for haptoglobin–hemoglobin complexes and is mostly expressed by monocytes and macrophages. Its levels are used to determine macrophage activation. We aimed to determine serum sCD163 in a sample of DLBCL patients and study eventual correlations with parameters of disease activity or survival. Serum sCD163 levels were measured in 40 frozen sera from patients diagnosed with DLBCL and 30 healthy individuals (HIs) using an enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed using SPSS version 28. The results showed that patients who achieved complete response after standard-of-care immunochemotherapy and were alive and disease-free after 12 months of follow-up but had elevated sCD163 levels (above median) at diagnosis presented a significantly worse overall survival compared to those with initial serum sCD163 levels below the median (p = 0.03). Consequently, serum sCD163 levels in patients with DLBCL may constitute a marker of long-term response to chemoimmunotherapy. [ABSTRACT FROM AUTHOR]

Simple Summary: The environment seems to first damage the skin allowing secondary bacteria to cause a local skin disease in bovines: digital dermatitis. Markers of inflammation, cytokines, are activated, but this does not initiate a systemic reaction and thus the body does not seem to be able to eliminate the disease. Digital dermatitis is a disease of the digital skin and causes lameness and welfare problems in dairy cattle. This study assessed the local and systemic inflammatory responses of cows with different digital dermatitis lesions and compared macroscopical and histological findings. Cow feet (n = 104) were evaluated macroscopically and skin biopsies histologically. Serum samples were analyzed for acute phase proteins (serum amyloid A and haptoglobin) and pro-inflammatory cytokines (interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha). Cows with macroscopically graded active lesions (p = 0.028) and non-active lesions (p = 0.008) had higher interleukin-1 beta levels in their serum compared to healthy cows. Interleukin-1 beta serum concentrations were also higher (p = 0.042) when comparing lesions with necrosis to lesions without necrosis. There was no difference when other cytokine or acute phase protein concentrations in healthy cows were compared to those in cows with different digital dermatitis lesions. A novel histopathological grading was developed based on the chronicity of the lesions and presence of necrosis and ulceration. The presence and number of spirochetes were graded separately. In the most severe chronic lesions, there was marked epidermal hyperplasia and hyperkeratosis with necrosis, deep ulceration, and suppurative inflammation. Spirochetes were found only in samples from necrotic lesions. This study established that digital dermatitis activates proinflammatory cytokines. However, this did not initiate the release of acute phase proteins from the liver. A histopathological grading that takes into account the age and severity of the lesions and presence of spirochetes was developed to better understand the progression of the disease. It is proposed that necrosis of the skin is a result of ischemic necrosis following reduced blood flow in the dermal papillae due to pressure and shear stress caused by thickened epidermis, and that the spirochetes are secondary invaders following tissue necrosis. [ABSTRACT FROM AUTHOR]

Simple Summary: There is an increasing interest in measuring biomarkers from saliva, as samples can be obtained noninvasively. This is particularly true in pigs where blood sampling is technically complicated and stressful. In pigs, studies regarding how the absorbent material used for collection affects analytical results are scarce. The aim of this study was to evaluate how the material used for saliva collection can affect the results of analyses for the different types of biomarkers that can be measured in the saliva of pigs. In this report, polypropylene sponges gave higher sample saliva volumes and, in some salivary analytes, provided different values compared with cotton rolls. Therefore, it can be concluded that the type of material used can influence the results of saliva analysis. The use of saliva as a biological sample from pigs is of high practical interest because blood collection from pigs is difficult and stressful. In this study, the influence of two different materials, a cotton roll and a polypropylene sponge, in porcine saliva collection was evaluated. For this purpose, the effect of the material used for sampling was evaluated in a panel of 13 analytes, including those related to stress (cortisol and oxytocin), inflammation and immunity (adenosine deaminase, haptoglobin and myeloperoxidase), redox homeostasis (the cupric reducing ability of saliva, the ferric reducing activity of saliva, and the Trolox equivalent antioxidant capacity), and sepsis (procalcitonin), as well as other routine analytes related to metabolism and different tissues and organs, such as lactate dehydrogenase, creatine kinase, urea, and total protein concentration. The polypropylene sponge provided a higher sample volume than the cotton roll. Although the results of some salivary analytes were equivalent for both materials, other analytes, such as creatine kinase, haptoglobin and total proteins, showed significant differences depending on the material used for saliva collection. Therefore, the type of material used for salivary collection in pigs should be considered when interpreting the results of analyses of the salivary analytes. [ABSTRACT FROM AUTHOR]

Haptoglobin is the body's first line of defence against the toxicity of extracellular haemoglobin released following a subarachnoid haemorrhage (SAH). We investigated the haptoglobin response after SAH in cerebrospinal fluid (CSF) and serum. Paired CSF and serum samples from 19 controls and 92 SAH patients were assayed as follows: ultra-performance liquid chromatography for CSF haemoglobin and haptoglobin, immunoassay for serum haptoglobin and multiplexed CSF cytokines, and colorimetry for albumin. There was marked CSF haptoglobin deficiency: 99% of extracellular haemoglobin was unbound. The quotients for both CSF/serum albumin (qAlb) and haptoglobin (qHp) were used to compute the CSF haptoglobin index (qHp/qAlb). CSF from SAH patients had a significantly lower haptoglobin index compared to controls, especially in Haptoglobin-1 allele carriers. Serum haptoglobin levels increased after SAH and were correlated with CSF cytokine levels. Haptoglobin variables were not associated with long-term clinical outcomes post-SAH. We conclude that: (1) intrathecal haptoglobin consumption occurs after SAH, more so in haptoglobin-1 allele carriers; (2) serum haptoglobin is upregulated after SAH, in keeping with the liver acute phase response to central inflammation; (3) haptoglobin in the CSF is so low that any variation is too small for this to affect long-term outcomes, emphasising the potential for therapeutic haptoglobin supplementation. [ABSTRACT FROM AUTHOR]

Rainbow trout, one of the most economically important aquaculture fish species worldwide, is affected by the pathogenic bacteria A. salmonicida, which causes furunculosis outbreaks, leading to huge economic losses. In this study, an oligonucleotide microarray was applied to identify transcriptional changes in the skin of rainbow trout individuals in response to a bacterial infection. Overall, 656 and 434 differentially expressed genes (DEGs) were identified at 2 and 6 days after a bacterial challenge (dpi), respectively. A comparison of moribund (2 dpi) and survivor fish (6 dpi) revealed 169 DEGs. Between these were many genes involved in immune response, including lysozymes, pattern recognition receptors (c-type lectins), antimicrobial peptides (cathelicidin and hepcidin), acute-phase proteins (serum amyloids and haptoglobin), complement cascade proteins (c3, c4, c6 and c7), interleukins (il11 and il1b) and chemokines (ccl19 and cxcl8). Alterations of leptin, eicosanoids and prostaglandins have been found, which suggest metabolic remodeling in conjunction with immune response. Further, the regulation of programmed cell death genes (caspase 8, bcl2 apoptosis regulator, nfkb inhibitor alpha and heme oxygenase) and structural proteins (collagens, myosins, keratins and metalloproteinases) was observed. This study provides, for the first time, a gene expression analysis of rainbow trout skin in response to A. salmonicida infection, revealing the complexity of defense strategies in response to furunculosis. [ABSTRACT FROM AUTHOR]

Background: Acute pancreatitis (AP) is a leading cause of gastrointestinal hospital admissions, with up to 40% mortality in patients with moderate–severe AP. Glycoprotein acetylation (GlycA) is measured as a nuclear magnetic resonance signal (NMR) of the post-translational modification of glycosylated acute-phase proteins released during inflammation. We aimed to investigate the role of GlycA as an inflammatory biomarker of AP. Methods: We prospectively enrolled 20 AP patients and 22 healthy controls and collected EDTA plasma samples at admission and discharge. NMR spectra were acquired from these samples using a 400 MHz Vantera® Clinical Analyzer, and GlycA concentrations were calculated (normal = 400 μmol/L). The GlycA NMR signal, at 2.00 ± 0.01 ppm in the NMR spectrum, is derived from the N-acetyl methyl group protons within the carbohydrate side chains of circulating glycoproteins such as α1-acid glycoprotein, haptoglobin, α1-antitrypsin, α1-antichymotrypsin, and transferrin. GlycA levels were then compared between AP patients and controls, as well as within the AP group, based on etiology and severity. Results: Demographic comparisons were similar, except for a higher BMI in AP patients compared to healthy controls (29.9 vs. 24.8 kg/m2; p < 0.001). AP was mild in 10 patients, moderate in 7, and severe in 3. GlycA levels were higher in AP patients than healthy controls on admission (578 vs. 376 μmol/L, p < 0.001) and at discharge (655 vs. 376 μmol/L, p < 0.001). GlycA levels were significantly higher in patients with moderate–severe AP than in those with mild AP at discharge (533 vs. 757 μmol/L, p = 0.023) but not at admission. After adjusting for BMI, multivariable regression indicated that patients with GlycA levels > 400 μmol/L had significantly higher odds of having AP of any severity (OR = 6.88; 95% CI, 2.07–32.2; p = 0.004) and mild AP (OR = 6.12; 95% CI, 1.48–42.0; p = 0.025) than controls. Conclusion: Our pilot study highlights the use of GlycA as a novel diagnostic biomarker of inflammation in patients with AP. Our study shows that GlycA levels were significantly higher in hospitalized AP patients compared to healthy controls. Patients with moderate-to-severe AP had higher GlycA levels compared to patients with mild AP at the time of their hospital discharge, suggesting persistent inflammation in patients with severe disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Weaning is one of the most stressful experiences for beef calves during their life cycle; however, it is a crucial practice for the production system. Calming pheromones can alleviate stress and reduce its negative impacts on behavior, growth, and the immune system. In this study, the administration of a bovine-appeasing substance enhanced body weight gain, reduced temperament scores and serum cortisol concentration, and improved the behavior and response to vaccination. Therefore, the bovine-appeasing substance could be used in complement to the best weaning practices to reduce stress and its impact on performance, temperament, grazing behavior, and the immune system. An analog of a bovine-appeasing substance (BAS) was previously demonstrated to have calming effects, and it could be an alternative to alleviate the stress caused by weaning. Thus, the objective of this study was to evaluate the effects of BAS administration at weaning on growth, stress, behavior, and response to vaccination of Nellore calves. Eighty-six Nellore calves (40 females and 46 males) were abruptly weaned and randomly assigned into 1 of 2 treatments: (1) saline solution (0.9% NaCl; n = 43) and (2) BAS (Secure Catte, IRSEA Group, Quartier Salignan, France; n = 43). The solutions were topically applied (5 mL/calf) to the nuchal skin area of each animal. On d 0, before treatment application, calves were vaccinated against infectious bovine rhinotracheitis (IBR), parainfluenza-3 (PI3) virus, and bovine viral diarrhea virus types 1 and 2 (BVDV-1 and 2). Calves from each treatment were kept in different pastures for 15 d (time of BAS action) and then moved to a single pasture. Body weight (BW), blood samples, and temperament in the chute (entry score, chute score, and exit score) were collected on d 0, 3, 8, 15, 51, and 100, and behavior on pasture on d 1, 2, 4, 5, 6, 7, and 9. Calves assigned to BAS vs. Saline treatment tended to have greater BW on d 15 (p = 0.10), tended to have lower entry scores on d 8 and 51 (p = 0.10), and chute scores on d 8 (p = 0.07), and had lower exit scores on d 8 (p = 0.02). Calves assigned to BAS vs. Saline treatment also had greater time grazing on d 7 and 9 (p < 0.01), eating concentrate on d 2, 5, and 6 (p = 0.05), walking on d 1, 2, 5, and 9 (p < 0.01), standing and ruminating on d 2, 7 and 9 (p < 0.01), and playing on d 2, 4, 6, 7, and 9 (p < 0.01). Furthermore, they had lower time lying on 1 and 2 (p < 0.01), standing on d 5 and 9 (p < 0.01), and vocalizing on d 1 and 2 (p < 0.01). Calves assigned to BAS vs. Saline treatment had greater serum titter concentrations of PI3 t on d 15 and 51 (p = 0.05) and BVDV-1 on d 51 (p = 0.02). However, they had lower serum concentrations of cortisol on d 3 (p = 0.03). BAS administration did not affect (p ≥ 0.12) the serum titer concentration of IBR and BVDV-2 titers or the plasma concentration of haptoglobin and ceruloplasmin. The BAS administration improved BW, reduced temperament and serum cortisol concentration, and improved behavior and response to vaccination. [ABSTRACT FROM AUTHOR]

Myocardial infarction (MI) with obstructive coronary artery disease (MI-CAD) and MI in the absence of obstructive coronary artery disease (MINOCA) affect different populations and may have separate pathophysiological mechanisms, with greater inflammatory activity in MINOCA compared to MI-CAD. Helicobacter pylori (Hp) can cause systemic inflammation and has been associated with cardiovascular disease (CVD). We aimed to investigate whether Hp infection is associated with concentrations of protein biomarkers of inflammation and CVD. In a case-control study, patients with MINOCA (n = 99) in Sweden were included, complemented by matched subjects with MI-CAD (n = 99) and controls (n = 100). Protein biomarkers were measured with a proximity extension assay in plasma samples collected 3 months after MI. The seroprevalence of Hp and cytotoxin-associated gene A (CagA) was determined using ELISA. The associations between protein levels and Hp status were studied with linear regression. The prevalence of Hp was 20.2%, 19.2%, and 16.0% for MINOCA, MI-CAD, and controls, respectively (p = 0.73). Seven proteins were associated with Hp in an adjusted model: tissue plasminogen activator (tPA), interleukin-6 (IL-6), myeloperoxidase (MPO), TNF-related activation-induced cytokine (TRANCE), pappalysin-1 (PAPPA), soluble urokinase plasminogen activator receptor (suPAR), and P-selectin glycoprotein ligand 1 (PSGL-1). Hp infection was present in one in five patients with MI, irrespective of the presence of obstructive CAD. Inflammatory proteins were elevated in Hp-positive subjects, thus not ruling out that Hp may promote an inflammatory response and potentially contribute to the development of CVD. [ABSTRACT FROM AUTHOR]

Simple Summary: Evaluations of transition cow nutrition strategies on health and performance in larger commercial farms are limited. In a 72-farm prospective cohort study, we evaluated the associations of common nutritional strategies fed during the far-off dry, close-up dry, and fresh periods with postpartum health and performance. Overall, our results support feeding a controlled energy diet prepartum and high-starch fresh diet to primiparous and multiparous cows. The objective was to identify relationships between transition cow nutritional strategies and the prevalence of elevated analytes (nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHB), and haptoglobin (Hp)), disorder incidence (DI), milk yield, and reproductive performance. Multiparous and primiparous cows from 72 farms in the northeastern US were enrolled in a herd-level cohort study. Farms were dichotomized within parity into a nutritional strategy within each period; far-off: controlled energy (CE; <16.5% starch and ≥40% forage neutral detergent fiber (FNDF)) or not CE (NCE; ≥16.5% starch or <40% FNDF or both), close-up: high FNDF (HF; ≥40% FNDF) or low FNDF (LF; <40% FNDF), and fresh: low starch (LS; <25.5% starch) or high starch (HS; ≥25.5% starch). No evidence existed that transition cow nutritional strategies were associated with milk yield outcomes (p ≥ 0.20). In general, our results support feeding multiparous cows HF close-up and HS fresh to minimize excessive BHB and DI; however, multiparous cows fed LF close-up had a higher pregnancy rate, and lower prepartum NEFA and Hp. Similarly, our results support feeding primiparous cows CE far-off, HF close-up, and HS fresh to maximize reproductive performance, and minimize BHB and DI; however, herds fed HF close-up or HS fresh had higher Hp. [ABSTRACT FROM AUTHOR]

The careful monitoring of patients with mild/moderate COVID-19 is of particular importance because of the rapid progression of complications associated with COVID-19. For prognostic reasons and for the economic management of health care resources, additional biomarkers need to be identified, and their monitoring can conceivably be performed in the early stages of the disease. In this retrospective cross-sectional study, we found that serum concentrations of high-mobility group box 1 (HMGB1) and heme oxygenase-1 (HO-1), at the time of hospital admission, could be useful biomarkers for COVID-19 management. The study included 160 randomly selected recovered patients with mild to moderate COVID-19 on admission. Compared with healthy controls, serum HMGB1 and HO-1 levels increased by 487.6 pg/mL versus 43.1 pg/mL and 1497.7 pg/mL versus 756.1 pg/mL, respectively. Serum HO-1 correlated significantly with serum HMGB1, oxidative stress parameters (malondialdehyde (MDA), the phosphatidylcholine/lysophosphatidylcholine ratio (PC/LPC), the ratio of reduced and oxidative glutathione (GSH/GSSG)), and anti-inflammatory acute phase proteins (ferritin, haptoglobin). Increased heme catabolism/hemolysis were not detected. We hypothesize that the increase in HO-1 in the early phase of COVID-19 disease is likely to have a survival benefit by providing protection against oxidative stress and inflammation, whereas the level of HMGB1 increase reflects the activity of the innate immune system and represents levels within which the disease can be kept under control. [ABSTRACT FROM AUTHOR]

Simple Summary: S100A8/A9 (also known as calprotectin) and S100A12 (also known as calgranulin C) are considered biomarkers of potential interest and are proteins members of the calgranulin family which is related to different inflammatory conditions, immune-mediated diseases, and sepsis. The objectives of this study were to evaluate if S100A8/A9 and A12 could change in the saliva of pigs with diarrhea due to E. coli and compare the changes of S100A8/A9 and A12 with other analytes in order to explore the possible causes or mechanisms involved in these changes. For this purpose, a panel integrated by other analytes related to inflammation, immune system, stress, tissue damage, sepsis, and redox status was also evaluated. S100A8/A9 and S100A12 increased in pigs with diarrhea produced by E. coli and were correlated with other salivary analytes. Further studies should be performed to elucidate the possible practical applications of these calgranulins as biomarkers to evaluate the health and welfare of pigs. The family of calgranulins includes S100A8 (calgranulin A), S100A9 (calgranulin B), which can appear as a heterodimer known as S100A8/A9 or calprotectin, and S100A12 (calgranulin C). These proteins are related to different inflammatory conditions, immune-mediated diseases, and sepsis and are considered biomarkers of potential interest. This study aims to evaluate if S100A8/A9 and A12 could change in pigs with diarrhea due to E. coli and to compare the changes of S100A8/A9 and A12 with other analytes in order to explore the possible causes or mechanisms involved. For this purpose, a panel integrated by analytes related to inflammation (haptoglobin, inter-alpha trypsin inhibitor 4 (ITIH4), and total protein); immune system (adenosine deaminase, ADA); stress (alpha-amylase); tissue damage (lactate and lactate dehydrogenase (LDH)); sepsis (aldolase) and redox status (ferric-reducing ability of saliva (FRAS) and advanced oxidation protein products (AOPP)) was evaluated. S100A8/A9 and A12 and the other analytes measured in this study showed increases in the saliva of pigs with diarrhea due to E. coli. S100A8/A9 and/or A12 showed a significant correlation of different magnitude with some of the other analytes evaluated. Further studies should be conducted to gain knowledge about the possible practical applications as biomarkers of the measurements of S100A8/A9 and A12 in the saliva of pigs. [ABSTRACT FROM AUTHOR]

Capsaicin (CAP) has various biological activities; it has antibacterial, anti-inflammatory and antioxidant properties, and stimulates intestinal development. The aim of this study was to investigate the effect of CAP on the health of nursing calves under group housing conditions. Twenty-four newborn Holstein calves were randomly assigned to three treatment groups of eight calves each. The milk replacer was supplemented with 0, 0.15 or 0.3 mL/d of CAP in each of the three treatment groups. Following a one-month clinical trial of individual-pen housing, an extended one-month trial of group housing was conducted. At the end of the trial, serum samples, rectal fecal samples and upper respiratory swab samples were collected to determine the effect of CAP addition on serum parameters, fecal fermentation parameters and upper respiratory microbiota of calves under group housing conditions. The results showed that the addition of high doses of CAP decreased calf respiratory scores (p < 0.05), increased serum glutathione peroxidase, superoxide dismutase, immunoglobulin A, immunoglobulin G, immunoglobulin M and interleukin-10 concentration (p < 0.05), and decreased malondialdehyde, amyloid A and haptoglobin concentration (p < 0.05). Moreover, high doses of CAP increased the rectal fecal concentration of total short-chain fatty acids, acetate and butyric acid (p < 0.05). In addition, CAP regulated the upper respiratory tract microbiota, with high doses of CAP reducing Mycoplasma abundance (p < 0.05), two doses of CAP reducing Corynebacterium abundance (p < 0.05) and a tendency to reduce Staphylococcus abundance (p = 0.06). Thus, CAP can improve calf antioxidant capacity, immune capacity and reduce inflammatory factors, stress proteins as well as improve gut fermentation and upper respiratory microbiota under group housing conditions, which is beneficial for healthy calf growth. [ABSTRACT FROM AUTHOR]

Simple Summary: Proteomics is a discipline investigating the proteins present in a specific biological environment, aiming to understand better the processes that take place in specific districts but also aiming for the possible discovery of new biomarkers. Herein, the fecal proteome of healthy dogs and dogs suffering from different hepatobiliary disorders was investigated. The study highlighted qualitative and quantitative differences between the groups of patients under analysis, leading us in particular to hypothesize a possible role for proteins such as fibronectin, haptoglobin, and trefoil factor 2. The present results need to be further confirmed by other tests and studies, but the direction taken appears promising. In the present study, the fecal proteomes of clinically healthy dogs (HD = n. 10), of dogs showing clinical, ultrasonographic, and/or laboratory evidence of different hepatobiliary dysfunction (DHD = n. 10), and of dogs suffering from chronic hepatitis (CHD = n. 10) were investigated with an Ultimate 3000 nanoUPLC system, coupled to an Orbitrap Fusion Lumos Tribrid mass spectrometer. Fifty-two different proteins of canine origin were identified qualitatively in the three study groups, and quantitative differences were found in 55 proteins when comparing groups. Quantitatively, a total of 41 and 36 proteins were found differentially abundant in the DHD and CHD groups compared to the control HD, and 38 proteins resulted dysregulated in the CHD group as compared to the DHD group. Among the various proteins, differently abundant fecal fibronectin and haptoglobin were more present in the feces of healthy and DHD dogs than in chronic ones, leading us to hypothesize its possible diagnostic/monitoring role in canine chronic hepatitis. On the other hand, the trefoil factor 2 was increased in DHD dogs. Our results show that the analysis of the fecal proteome is a very promising field of study, and in the case of dogs suffering from different hepatobiliary disorders, it was able to highlight both qualitative and quantitative differences among the three groups included. Results need to be confirmed with western blotting and in further studies. [ABSTRACT FROM AUTHOR]

This work proposes an outstanding screening magneto-bioassay for the early identification of bovine subclinical mastitis. Haptoglobin (Hp) was used as a promising biomarker for the diagnosis of subclinical mastitis. To this end, novel chitosan-modified magnetic particles, coated with haemoglobin-modified polyaniline (MNPs@Chi/PANI-Hb), have been integrated in a sensitive electrochemical Hp bioassay. Haemoglobin was used as bioreceptor due to its high affinity against Hp. The appropriateness of their synthesis and their modifications were demonstrated by XRD, FT-IR spectroscopy, thermogravimetry and N2 adsorption–desorption analyses. After the optimization of the experimental parameters the main analytical features were obtained showing excellent performance. The electrochemical biosensor in milk matrix presented a dynamic range of 0.001 to 0.32 μg mL−1 with a detection limit of 0.031 μg mL−1, a value much lower than the normal Hp values in bovine milk, making it highly suitable for such determinations. Finally, real milk samples, obtained from local dairy farmers, were analysed by the electrochemical bioassay and compared against a commercial ELISA kit for Hp detection. Milk samples were correctly classified as "acceptable" or "unacceptable" milk considering the Hp concentration obtained with the proposed bioassay, confirming its excellent predictive capacity for subclinical mastitis diagnosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Analysis and classification of effusions is helpful in guiding diagnostic tests to identify an underlying disease. The traditional veterinary classification of effusions does not adequately reflect the disease mechanism causing the effusion in cats and might not sufficiently inform diagnostic work-up. The aim of this study was to assess whether other parameters might aid classification. Sixty-five cats with body cavity effusions (within the abdomen, chest and pericardial sac) were included. Effusions were classified as transudates (e.g., heart failure [n = 18]) or exudates (e.g., inflammation, cancer [n = 47]) based on the disease mechanism causing the effusion, and using the traditional scheme. Several parameters (activity of enzyme lactate dehydrogenase in the effusion; effusion/serum ratio of the enzymatic activity of lactate dehydrogenase; effusion/serum total protein ratio; serum–effusion albumin gradient; acute phase proteins in serum and effusion) were analysed in their ability to differentiate exudates from transudates. All tested parameters performed better in classifying effusions in comparison to the traditional scheme. Acute phase proteins were helpful in classifying effusions based on their disease mechanism but could not separate effusion types when using the traditional scheme. This latter finding further supports the classification of effusions based on their disease mechanism. Better classification of effusions might improve disease diagnosis in the future. The traditional veterinary classification (TVC) of effusions based on cell count and total protein (TP) does not adequately reflect the aetiology. Light's criteria (LC) (activity of lactate dehydrogenase [LDH] in the effusion [LDHef], effusion/serum LDH ratio [LDHr], effusion/serum TP ratio [TPr]), serum–effusion albumin gradient (ALBg), acute phase proteins (APPs) [serum amyloid A (SAA), α1-acid glycoprotein (AGP), haptoglobin] might aid classification. The aim was to evaluate the utility of these parameters except LDHr in differentiating exudates from transudates. Sixty-five cats with effusions (33 peritoneal, 31 pleural, 1 pericardial), with 18 transudates and 47 exudates based on aetiological classification (AC), were included. The sensitivity, specificity and accuracy of several parameters to identify exudates (based on AC) was assessed. APPs were compared between exudates and transudates based on AC and TVC, with receiver operating characteristics analysis identifying the best APP to recognise exudates. Simplified LC (LDHef, TPr) had an accuracy of 79% and TVC of 48%. ALBg had the highest sensitivity (98%) and LDHef the highest specificity (83%) in identifying exudates in cats. All APPs but effusion SAA could differentiate exudates from transudates based on AC (effusion AGP had the largest area under the curve 0.79) but not TVC. All parameters were better than TVC in identifying exudates. The conformity of APPs with AC but not TVC favours the use of AC to classify effusions. [ABSTRACT FROM AUTHOR]

A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using various experimental methods: liquid chromatography–mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic-force microscopy. Infrared spectroscopy found no significant difference between the tear fluid of MS patients and the control spectra; all three significant peaks were located at around the same positions. Raman analysis showed differences between the spectra of the tear fluid of MS patients and the spectra of healthy subjects, which indicated a decrease in tryptophan and phenylalanine content and changes in the relative contributions of the secondary structures of the polypeptide chains of tear proteins. Atomic-force microscopy exhibited a surface fern-shaped dendrite morphology of the tear fluid of patients with MS, with less roughness on both oriented silicon (100) and glass substrates compared to the tear fluid of control subjects. The results of liquid chromatography–mass spectrometry showed downregulation of glycosphingolipid metabolism, sphingolipid metabolism, and lipid metabolism. Proteomic analysis identified upregulated proteins in the tear fluid of patients with MS such as cystatine, phospholipid transfer protein, transcobalamin-1, immunoglobulin lambda variable 1–47, lactoperoxidase, and ferroptosis suppressor protein 1; and downregulated proteins such as haptoglobin, prosaposin, cytoskeletal keratin type I pre-mRNA-processing factor 17, neutrophil gelatinase-associated lipocalin, and phospholipase A2. This study showed that the tear proteome in patients with MS is modified and can reflect inflammation. Tear fluid is not a commonly used biological material in clinico-biochemical laboratories. Experimental proteomics has the potential to become a promising contemporary tool for personalized medicine, and it might be applied in clinical practice by providing a detailed analysis of the tear-fluid proteomic profile of patients with MS. [ABSTRACT FROM AUTHOR]

Canine myxomatous mitral valve disease (MMVD) is similar to Barlow's form of MMVD in humans. These valvulopathies are complex, with varying speeds of progression. We hypothesized that the relative abundances of serum proteins would help identify the consecutive MMVD stages and discover new disease pathways on a systemic level. To identify distinction-contributing protein panels for disease onset and progression, we compared the proteomic profiles of serum from healthy dogs and dogs with different stages of naturally occurring MMVD. Dogs were divided into experimental groups on the basis of the left-atrium-to-aorta ratio and normalized left ventricular internal dimension in diastole values. Serum was collected from healthy (N = 12) dogs, dogs diagnosed with MMVD in stages B1 (N = 13) and B2 (N = 12) (asymptomatic), and dogs diagnosed with MMVD in chronic stage C (N = 13) (symptomatic). Serum biochemistry and selected ELISAs (galectin-3, suppression of tumorigenicity, and asymmetric dimethylarginine) were performed. Liquid chromatography–mass spectrometry (LC–MS), tandem mass tag (TMT) quantitative proteomics, and statistical and bioinformatics analysis were employed. Most of the 21 serum proteins with significantly different abundances between experimental groups (p < 0.05, FDR ˂ 0.05) were classified as matrix metalloproteinases, protease inhibitors, scaffold/adaptor proteins, complement components, anticoagulants, cytokine, and chaperone. LC–MS TMT proteomics results obtained for haptoglobin, clusterin, and peptidase D were further validated analytically. Canine MMVD stages, including, for the first time, asymptomatic B1 and B2 stages, were successfully distinguished in dogs with the disease and healthy dogs on the basis of the relative abundances of a panel of specific serum proteins. Most proteins with significantly different abundances were involved in immune and inflammatory pathways. Their role in structural remodeling and progression of canine MMVD must be further investigated. Further research is needed to confirm the resemblance/difference with human MMVD. Proteomics data are available via ProteomeXchange with the unique dataset identifier PXD038475. [ABSTRACT FROM AUTHOR]

Measurements of specific acute phase proteins (APP) and protein electrophoresis (EPH) fractions have been widely used to better assess the health of species under managed care across numerous taxa. To date, APP assays have not been validated in the hamadryas baboon (Papio hamadryas), and reference intervals have not yet been established. This information is critical for the interpretation of APP and EPH measurements used in the diagnosis of inflammatory diseases during routine veterinary care of this species. To obtain this information, banked serum samples from hamadryas baboons of various age, sex, and health status, under managed care at the North Carolina Zoo, were analyzed. A small pilot study found significantly higher serum amyloid A (SAA) and C-reactive protein (CRP) but not haptoglobin in baboons with acute inflammation compared to healthy counterparts, so these two APPs were investigated further. Reagents for serum amyloid A (SAA) and C-reactive protein (CRP) were validated, although differences in CRP reagents were observed. Based on the results of this study, SAA and CRP were defined as major APPs that were significantly increased in baboons with active inflammation or infection compared to healthy conspecifics. Baboons with acute inflammation additionally had significantly higher gamma globulins compared to healthy baboons. Although mean albumin concentrations were lower in baboons with acute inflammation, the difference from healthy baboons was not statistically significant. This study identifies SAA, CRP, and EPH as useful tools in the diagnosis of inflammatory disease in the hamadryas baboon and establishes reference intervals to aid in the future veterinary care of this species. [ABSTRACT FROM AUTHOR]

Heme is the reactive center of several metal-based proteins that are involved in multiple biological processes. However, free heme, defined as the labile heme pool, has toxic properties that are derived from its hydrophobic nature and the Fe-atom. Therefore, the heme concentration must be tightly controlled to maintain cellular homeostasis and to avoid pathological conditions. Therefore, different systems have been developed to scavenge either Hb (i.e., haptoglobin (Hp)) or the free heme (i.e., high-density lipoproteins (HDL), low-density lipoproteins (LDL), hemopexin (Hx), and human serum albumin (HSA)). In the first seconds after heme appearance in the plasma, more than 80% of the heme binds to HDL and LDL, and only the remaining 20% binds to Hx and HSA. Then, HSA slowly removes most of the heme from HDL and LDL, and finally, heme transits to Hx, which releases it into hepatic parenchymal cells. The Hx:heme or HSA:heme complexes are internalized via endocytosis mediated by the CD91 and CD71 receptors, respectively. As heme constitutes a major iron source for pathogens, bacteria have evolved hemophores that can extract and uptake heme from host proteins, including HSA:heme. Here, the molecular mechanisms underlying heme scavenging and delivery from HSA are reviewed. Moreover, the relevance of HSA in disease states associated with increased heme plasma concentrations are discussed. [ABSTRACT FROM AUTHOR]

Intense exercise can cause inflammation and oxidative stress due to the production of reactive oxygen species. These pathophysiological processes are interdependent, and each one can induce the other, creating a vicious circle. A placebo-controlled blind study was carried out in show jumping horses (n. 16) to evaluate the effects of a commercial dietary supplement (Dolhorse® N.B.F. Lanes srl, Milan, Italy) containing Verbascum thapsus leaf powder (1.42%), Curcuma longa (14.280 mg/kg), and Boswellia serrata (Roxb ex Colebr) (14.280 mg/kg) extracts. Before and after 10 days of dietary supplementation, blood samples were collected to evaluate the protein levels, antioxidants, and inflammatory responses by proteomic analysis or real-time Reverse Transcriptase-Polymerase Chain Reaction (real-time RT-PCR). A total of 36 protein spots, connected to 29 proteins, were modulated by dietary supplementation, whereas real-time RT-PCR revealed a significant downregulation of proinflammatory cytokines (interleukin 1α (p < 0.05) and interleukin-6 (0.005), toll-like receptor 4 (p < 0.05), and IKBKB (p < 0.05) in supplemented sport horses. Immunoglobulin chains, gelsolin, plasminogen, vitamin D binding protein, apolipoprotein AIV, and filamin B were overexpressed, whereas haptoglobin, α-2-HS-glycoprotein, α2-macroglobulin, afamin, amine oxidase, 60S acidic ribosomal protein, and complement fragments 3, 4, and 7 were reduced. No effect was observed on the antioxidant defense systems. The present results suggest this phytotherapy may reinforce the innate immune responses, thus representing a valid adjuvant to alleviate inflammation, which is a pathophysiological process in sport horses. [ABSTRACT FROM AUTHOR]

Hypertension and diabetes induce vascular injury through processes that are not fully understood. Changes in extracellular vesicle (EV) composition could provide novel insights. Here, we examined the protein composition of circulating EVs from hypertensive, diabetic and healthy mice. EVs were isolated from transgenic mice overexpressing human renin in the liver (TtRhRen, hypertensive), OVE26 type 1 diabetic mice and wild-type (WT) mice. Protein content was analyzed using liquid chromatography–mass spectrometry. We identified 544 independent proteins, of which 408 were found in all groups, 34 were exclusive to WT, 16 were exclusive to OVE26 and 5 were exclusive to TTRhRen mice. Amongst the differentially expressed proteins, haptoglobin (HPT) was upregulated and ankyrin-1 (ANK1) was downregulated in OVE26 and TtRhRen mice compared with WT controls. Conversely, TSP4 and Co3A1 were upregulated and SAA4 was downregulated exclusively in diabetic mice; and PPN was upregulated and SPTB1 and SPTA1 were downregulated in hypertensive mice, compared to WT mice. Ingenuity pathway analysis identified enrichment in proteins associated with SNARE signaling, the complement system and NAD homeostasis in EVs from diabetic mice. Conversely, in EVs from hypertensive mice, there was enrichment in semaphroin and Rho signaling. Further analysis of these changes may improve understanding of vascular injury in hypertension and diabetes. [ABSTRACT FROM AUTHOR]

Reproductive and condition parameters' dependency on immune status in seasonally reproducing ruminants such as red deer have not been outlined to date. We determined T and B blood lymphocytes; the concentration of IgG, cAMP, haptoglobulin, and 6-keto-PGF1α in blood plasma; and the mRNA and protein expression of PG endoperoxide synthase 2, 5-lipoxygenase, PGE2 synthase (PGES), PGF2α synthase (PGFS), PGI2 synthase (PGIS), leukotriene (LT)A4 hydrolase, and LTC4 synthase (LTC4S) in the uterine endo- and myometrium, on the 4th (N = 7) and 13th (N = 8) days of the estrous cycle, in anestrus (N = 6) and pregnancy (N = 8) in hinds. An increase in CD4+ T regulatory lymphocyte percentage during the estrous cycle and anestrus compared with pregnancy was recorded; the opposite effect was observed for CD21+ B cells (p < 0.05). cAMP and haptoglobin concentration were elevated during the cycle, as was IgG on the fourth day of the cycle, whereas 6-keto-PGF1α concentration was the highest in pregnancy, and the nearest in anestrus similarly were LTC4S, PGES, PGFS, and PGIS protein expression in the endometrium (p < 0.05). We showed an interaction between the immune system activation and AA-metabolite production in the uterus throughout different reproductive stages. IgG, cAMP, haptoglobin, and 6-keto-PGF1α concentrations are valuable candidates for markers of reproductive status in hinds. The results help expand our knowledge of the mechanisms underlying seasonal reproduction in ruminants. [ABSTRACT FROM AUTHOR]

The purpose of this trial was to evaluate serum levels of oxidative stress biomarkers and biochemical analytes in crossbred lambs during the rearing phase in an integrated crop-livestock system (ICLS) to control gastrointestinal parasites. The experiment used 36 crossbred lambs (cross: Ile de France × White Dorper × Texel) divided into two groups. The WCS group was supplemented with whole cottonseed (WCS), and controls had no supplementation. Body weight, blood collection, and fecal analysis of nematode eggs and Eimeria oocysts counting per gram of feces were performed for each animal within 84 days of experiment. The following serum analytes were determined: total protein, albumin, globulin, cholesterol, haptoglobin, and 10 oxidative stress biomarkers: cupric reducing antioxidant capacity, ferric reducing ability of plasma, trolox equivalent antioxidant capacity, thiol, uric acid, paraoxonase-1, total oxidant status, ferric-xylenol orange, advanced oxidation protein products, and reactive oxygen metabolites derived compounds. The inclusion of WCS suggested the benefit in controlling infection as well as inducing an increase in antioxidants and a decrease in oxidants in lambs naturally infected by gastrointestinal parasites. The combination of WCS and ICLS could be a useful tool in controlling gastrointestinal parasite infection without affecting the production performance. [ABSTRACT FROM AUTHOR]

Dairy cows with ketosis have high circulating beta-hydroxybutyric acid (BHBA) concentrations alongside which inflammation is concomitantly developed. Tryptophan (Trp) is an essential amino acid that participates in the regulation of the inflammatory response. However, the association between Trp metabolism and inflammation in dairy cows with ketosis remains unclear. Therefore, blood samples from healthy (n = 10) and ketotic (n = 10) primiparous dairy cows were collected at the calving date and the day of ketosis diagnosis (7 days in milk (7 DIM)). Serum levels of non-esterified fatty acids (NEFA), BHBA, haptoglobin (HP), serum amyloid A (SAA), lipopolysaccharide, and cortisol were analyzed. Tryptophan and its metabolites were quantified using liquid chromatography–tandem mass spectrometry. At 7 DIM, the concentrations of NEFA, BHBA, HP, and SAA were higher and the levels of Trp, kynurenine (KYN), indoleacetic acid, indole-3-lactic acid, and 3-indoxyl sulfate were lower in the dairy cows with ketosis compared with those in the healthy cows. However, the KYN/Trp and melatonin/Trp ratios increased in the cows with ketosis. At the calving date, the serum lipopolysaccharide levels did not differ between the healthy and ketotic cows, whereas the levels of NEFA, HP, and cortisol increased in the ketotic cows. Correlation analysis showed that Trp deficiency and elevated Trp metabolism in the dairy cows occurred during ketosis. Overall, our results suggest that abnormal Trp metabolism may contribute to the pathogenesis of ketosis. [ABSTRACT FROM AUTHOR]

Producers of milk and dairy products have been faced with the challenge of responding to European society's demand for guaranteed animal welfare production. In recent years, measures have been taken to improve animal welfare conditions on farms and evaluation systems have been developed to certify them, such as the Welfare Quality® protocol. Among the markers used for this purpose, acute phase proteins stand out, with haptoglobin being one of the most relevant. However, the diagnostic power of these tools is limited and more sensitive and specific technologies are required to monitor animal health status. Different factors such as diet, stress, and diseases modify the metabolism of the animals, altering the composition of the milk in terms of oligosaccharides, proteins, and lipids. Thus, in order to study oxidative-stress-associated lipids, a collection of well-characterized milk samples, both by veterinary diagnosis and by content of the acute stress biomarker haptoglobin, was analyzed by mass spectrometry and artificial intelligence. Two lipid species (sphingomyelin and phosphatidylcholine) were identified as potential biomarkers of health status in dairy cows. Both lipids allow for the discrimination of milk from sick animals and also milk from those with stress. Moreover, lipidomics revealed specific lipid profiles depending on the origin of the samples and the degree of freedom of the animals on the farm. These data provide evidence for specific lipid changes in stressed animals and open up the possibility that haptoglobin could also affect lipid metabolism in cow's milk. [ABSTRACT FROM AUTHOR]

The glycosylation of proteins plays an important role in neurological development and disease. Glycoproteomic studies on cerebrospinal fluid (CSF) are a valuable tool to gain insight into brain glycosylation and its changes in disease. However, it is important to consider that most proteins in CSFs originate from the blood and enter the CSF across the blood–CSF barrier, thus not reflecting the glycosylation status of the brain. Here, we apply a glycoproteomics method to human CSF, focusing on differences between brain- and blood-derived proteins. To facilitate the analysis of the glycan site occupancy, we refrain from glycopeptide enrichment. In healthy individuals, we describe the presence of heterogeneous brain-type N-glycans on prostaglandin H2-D isomerase alongside the dominant plasma-type N-glycans for proteins such as transferrin or haptoglobin, showing the tissue specificity of protein glycosylation. We apply our methodology to patients diagnosed with various genetic glycosylation disorders who have neurological impairments. In patients with severe glycosylation alterations, we observe that heavily truncated glycans and a complete loss of glycans are more pronounced in brain-derived proteins. We speculate that a similar effect can be observed in other neurological diseases where a focus on brain-derived proteins in the CSF could be similarly beneficial to gain insight into disease-related changes. [ABSTRACT FROM AUTHOR]

Adipose tissue is an endocrine organ that interferes with the severity of chronic obstructive pulmonary disease (COPD). Although inflammatory markers, body composition, and nutritional status have a significant impact on pulmonary function, the real contribution of adipocytokines and myokines in COPD is still controversial. We aimed to evaluate the role played by the body composition, leptin, adiponectin, haptoglobin, and irisin on the functional exercise capacity, respiratory function, and quality of life (QoL) in COPD. In 25 COPD (20% GOLD-1; 60% GOLD-2; 20% GOLD-3) patients and 26 matched control subjects, we find that leptin, total adiponectin and haptoglobin are significantly increased whereas the 6 min walk test (6MWT) and physical functioning scores are significantly decreased in COPD versus controls. A significant positive relationship is found between leptin and fat mass and between 6MWT and the good health indicators of nutritional status. A significant inverse relationship is found between 6MWT and leptin and fat mass, FEV1 and haptoglobin, and irisin and haptoglobin. Phase angle and leptin level are significant predictors for functional exercise capacity assessed with 6MWT. Taken altogether, the results of this pilot study further support the role played by body composition and adipocytokines on exercise capacity respiratory function and QoL in COPD. [ABSTRACT FROM AUTHOR]

The main cause of Parkinson's disease (PD) remains unknown and the pathologic changes in the brain limit rapid diagnosis. Herein, differentially expressed genes (DEGs) in the Gene Expression Omnibus (GEO) database (GSE8397 and GSE22491) were assessed using linear models for microarray analysis (limma). Ankyrin 1 (ANK1) was the only common gene differentially down-regulated in lateral substantia nigra (LSN), medial substantia nigra (MSN) and blood. Additionally, DEGs between high ANK1 and low ANK1 in GSE99039 were picked out and then uploaded to the Database for Annotation, Visualization and Integrated Discovery (DAVID) for gene ontology (GO) functional annotation analysis. GO analysis displayed that these DEGs were mainly enriched in oxygen transport, myeloid cell development and gas transport (biological process (BP)); hemoglobin complex, haptoglobin–hemoglobin complex and cortical cytoskeleton (cellular component (CC)); and oxygen transporter activity, haptoglobin binding and oxygen binding (molecular function (MF)). Receiver operating characteristic (ROC) curve analysis showed ANK1 had good diagnostic accuracy and increased the area under the curve (AUC) value when combined with other biomarkers. Consistently, intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropy-ridi-ne (MPTP) in C57BL/6J mice reduced ANK1 mRNA expression in both substantia nigra and blood compared to the control group. Thus, ANK1 may serve as a candidate biomarker for PD diagnosis. [ABSTRACT FROM AUTHOR]

Delayed cerebral ischemia (DCI) and vasospasm are two complications of subarachnoid hemorrhages (SAHs) which entail high risks of morbidity and mortality. However, it is unknown why only some patients who suffer SAHs will experience DCI and vasospasm. The purpose of this review is to describe the main genetic single nucleotide polymorphisms (SNPs) that have demonstrated a relationship with these complications. The SNP of the nitric oxide endothelial synthase (eNOS) has been related to the size and rupture of an aneurysm, as well as to DCI, vasospasm, and poor neurological outcome. The SNPs responsible for the asymmetric dimetilarginine and the high-mobility group box 1 have also been associated with DCI. An association between vasospasm and the SNPs of the eNOS, the haptoglobin, and the endothelin-1 receptor has been found. The SNPs of the angiotensin-converting enzyme have been related to DCI and poor neurological outcome. Studies on the SNPs of the Ryanodine Receptor yielded varying results regarding their association with vasospasm. [ABSTRACT FROM AUTHOR]

To study the associations of blood proteins with the presence of unstable atherosclerotic plaques in the arteries of patients with coronary atherosclerosis using quantitative proteomics. The studies involved two groups of men with coronary atherosclerosis (group 1 (St) had only stable atherosclerotic plaques; group 2 (Ns) had only unstable atherosclerotic plaques, according to histological analysis of tissue samples); the average age of patients was 57.95 ± 7.22. Protein concentrations in serum samples were determined using the PeptiQuant Plus Proteomics Kit. The identification of protein fractions was carried out by monitoring multiple reactions on a Q-TRAP 6500 mass spectrometer combined with a liquid chromatograph. Mass spectrometric identification revealed in serum samples from patients with unstable atherosclerotic plaques a reduced concentration of proteins in the blood: α-1-acid glycoprotein, α-1-antichymotrypsin, α-1-antitrypsin, ceruloplasmin, hemopexin, haptoglobin, apolipoprotein B-100, apolipoprotein L1, afamin and complement component (C3, C7, C9). Moreover, at the same time a high concentration complements factor H and attractin. The differences were considered significant at p < 0.05. It was found that the instability of atherosclerotic plaques is associated with the concentration of proteins: afamin, attractin, components of the complement system, hemopexin and haptoglobin. The data of our study showed the association of some blood proteins with the instability of atherosclerotic plaques in coronary atherosclerosis. Their potential role in the development of this disease and the possibility of using the studied proteins as biomarkers requires further research. [ABSTRACT FROM AUTHOR]

Aberrations in blood cells are common among heavy alcohol drinkers. In order to shed further light on such responses, we compared blood cell status with markers of hemolysis, mediators of inflammation and immune responses to ethanol metabolites in alcohol-dependent patients at the time of admission for detoxification and after abstinence. Blood cell counts, indices of hemolysis (LDH, haptoglobin, bilirubin), calprotectin (a marker of neutrophil activation), suPAR, CD163, pro- and anti-inflammatory cytokines and autoantibodies against protein adducts with acetaldehyde, the first metabolite of ethanol, were measured from alcohol-dependent patients (73 men, 26 women, mean age 43.8 ± 10.4 years) at baseline and after 8 ± 1 days of abstinence. The assessments also included information on the quantities of alcohol drinking and assays for biomarkers of alcohol consumption (CDT), liver function (AST, ALT, ALP, GGT) and acute phase reactants of inflammation. At baseline, the patients showed elevated values of CDT and biomarkers of liver status, which decreased significantly during abstinence. A significant decrease also occurred in LDH, bilirubin, CD163 and IgA and IgM antibodies against acetaldehyde adducts, whereas a significant increase was noted in blood leukocytes, platelets, MCV and suPAR levels. The changes in blood leukocytes correlated with those in serum calprotectin (p < 0.001), haptoglobin (p < 0.001), IL-6 (p < 0.02) and suPAR (p < 0.02). The changes in MCV correlated with those in LDH (p < 0.02), MCH (p < 0.01), bilirubin (p < 0.001) and anti-adduct IgG (p < 0.01). The data indicates that ethanol-induced changes in blood leukocytes are related with acute phase reactants of inflammation and release of neutrophil calprotectin. The studies also highlight the role of hemolysis and immune responses to ethanol metabolites underlying erythrocyte abnormalities in alcohol abusers. [ABSTRACT FROM AUTHOR]

Obesity and pregnancy may have synergistic effects on periodontal condition, and proteomics could be an ideal approach to highlight the pathophysiological mechanisms associated with these outcomes. This study analyzed the salivary proteomics related to obesity and periodontitis in women during pregnancy (T1) and after delivery (T2). Initially, 126 women were recruited and forty were allocated into groups: with obesity and periodontitis (OP); with obesity, but without periodontitis (OWP); with normal BMI, but with periodontitis (NP); with normal BMI and without periodontitis (NWP). Whole-mouth saliva was collected in T1 and T2, and proteins were extracted and individually processed by label-free proteomics (nLC-ESI-MS/MS). The up-regulations of Heat shock 70 kDa protein 1A, 1B, and 1-like were related to both obesity and periodontitis, separately. Albumin and Thioredoxin were up-regulated in periodontitis cases, while Cystatins (mainly S, SA, SN) and Lactotransferrin were down-regulated. The high abundances of Submaxillary gland androgen-regulated protein 3B, Protein S100-A8, Matrix metalloproteinase-9, Heat shock 70 kDa protein 2 and 6, Putative Heat shock 70 kDa protein 7, Heat shock 71 kDa protein, Haptoglobin and Plastin-1 were significant in the combination of obesity and periodontitis. Obesity and periodontitis remarkably altered the proteome of the saliva during pregnancy with substantial alterations after delivery. [ABSTRACT FROM AUTHOR]

Gold nanoclusters have revealed great potential as nanoantibiotics due to their superior chemical and physical characteristics. In this study, a peptide with 83 amino acids derived from haptoglobin was utilized as a surface ligand to synthesize gold nanoclusters via a facile hydrothermal approach. Characterization of the structural and optical properties demonstrated the successful synthesis of derived haptoglobin-conjugated gold nanoclusters. The spherical derived haptoglobin-conjugated gold nanoclusters exhibited a (111) plane of cubic gold and an ultra-small size of 3.6 ± 0.1 nm. The optical properties such as ultraviolet-visible absorption spectra, X-ray photoelectron spectroscopy spectra, fluorescence spectra, and Fourier transform infrared spectra also validated the successful conjugation between the derived haptoglobin peptide and the gold nanoclusters surface. The antibacterial activity, reactive oxygen species production, and antibacterial mechanisms of derived haptoglobin-conjugated gold nanoclusters were confirmed by culturing the bacterium Escherichia coli with hemoglobin to simulate bacteremia. The surface ligand of the derived haptoglobin peptide of derived haptoglobin-conjugated gold nanoclusters was able to conjugate with hemoglobin to inhibit the growth of Escherichia coli. The derived haptoglobin-conjugated gold nanoclusters with an ultra-small size also induced reactive oxygen species production, which resulted in the death of Escherichia coli. The superior antibacterial activity of derived haptoglobin-conjugated gold nanoclusters can be attributed to the synergistic effect of the surface ligand of the derived haptoglobin peptide and the ultra-small size. Our work demonstrated derived haptoglobin-conjugated gold nanoclusters as a promising nanoantibiotic for combating bacteremia. [ABSTRACT FROM AUTHOR]

We have recently shown that the blood cell-derived secretome of remote ischaemic (RIC)-conditioned individuals provides an external source of neuroprotection. In this study, we identified the bioactive compounds from the total proteins released by those cells. Our main strategy was to separate protein–protein complexes while maintaining their native structure and testing their bioactive properties. Subsequently, we identified up- and downregulated bioactive proteins. We uncovered two bioactive fractions composed of 18 proteins. Most of the protein peaks were unchanged; however, RIC mediated a decrease in two peaks (comprising seven proteins) and an increase in one peak (identified as haptoglobin). When focussing on the biological activity of these proteins, we found positive impacts on the regulation of cellular metabolic processes and an increase in biological processes related to the acute phase response and inflammation in the RIC-treated samples. Although we have identified the 18 proteins that exert the greatest cytoprotection, additional studies are needed to elucidate their particular function and detailed mechanisms of action. [ABSTRACT FROM AUTHOR]

Intracerebral hemorrhage (ICH) is recognized as a severe clinical problem lacking effective treatment. High mobility group box-1 (HMGB1) exhibits inflammatory cytokine-like activity once released into the extracellular space from the nuclei. We previously demonstrated that intravenous injection of rat anti-HMGB1 monoclonal antibody (mAb) remarkably ameliorated brain injury in a rat ICH model. Therefore, we developed a humanized anti-HMGB1 mAb (OKY001) for clinical use. The present study examined whether and how the humanized anti-HMGB1 mAb ameliorates ICH injury in common marmosets. The results show that administration of humanized anti-HMGB1 mAb inhibited HMGB1 release from the brain into plasma, in association with a decrease of 4-hydroxynonenal (4-HNE) accumulation and a decrease in cerebral iron deposition. In addition, humanized anti-HMGB1 mAb treatment resulted in a reduction in brain injury volume at 12 d after ICH induction. Our in vitro experiment showed that recombinant HMGB1 inhibited hemoglobin uptake by macrophages through CD163 in the presence of haptoglobin, suggesting that the release of excess HMGB1 from the brain may induce a delay in hemoglobin scavenging, thereby allowing the toxic effects of hemoglobin, heme, and Fe2+ to persist. Finally, humanized anti-HMGB1 mAb reduced body weight loss and improved behavioral performance after ICH. Taken together, these results suggest that intravenous injection of humanized anti-HMGB1 mAb has potential as a novel therapeutic strategy for ICH. [ABSTRACT FROM AUTHOR]

(1) Background: This study evaluated the effects of a plant bioactive (Phyto Ax'Cell, Phytosynthese, Mozac, France) on the inflammatory status and health of dairy cows during calving. (2) Methods: 46 Holstein crossbred cows were randomized into a control group (CON, n = 23) and the Phyto Ax'Cell group (PAC, n = 23). PAC received Phyto Ax'Cell at 25 g/cow/day, from 15 days prepartum to 7 days postpartum. Blood analyses were performed weekly from D-7 to D14 to evaluate the energy metabolism and inflammatory status; rectal temperature was measured daily within 14 days from calving day (D0). (3) Results: PAC showed lower serum haptoglobin at D7 (0.55 vs. 0.79 mg/mL; p < 0.05) and D14 (0.44 vs. 0.66 mg/mL; p < 0.05). CON had a higher number of circulating white blood cells and granulocytes on D7 (p < 0.05). Fewer cows from PAC showed hyperthermia (≥39 °C) during the first 2 weeks postpartum (−7%, p < 0.05). Energy metabolism, which was represented by the NEFA/cholesterol ratio, improved (0.21 vs. 0.36 at D0, p < 0.1; 0.19 and 0.15 vs. 0.36 and 0.32, respectively, at D+7 and D+14, p < 0.05) under the plant bioactive supplementation. (4) Conclusions: The results suggest that the anti-inflammatory plant bioactive compound with Brazilian green propolis administered during calving had a beneficial effect on the energy and inflammatory status of dairy cows. [ABSTRACT FROM AUTHOR]

Background: Angiotensin-converting enzyme 2 (ACE2) is implicated as a host cell receptor that causes infection in the pathogenesis of coronavirus disease 2019 (COVID-19), and its genetic polymorphisms in the ACE2 gene may promote cardiovascular disease and systemic inflammatory injury in COVID-19 patients. Hence, the genetic background may potentially explain the broad interindividual variation in disease susceptibility and/or severity. Methods: Genetic susceptibility to COVID-19 was analyzed by examining single-nucleotide polymorphisms (SNPs) of ACE2 in 246 patients with COVID-19 and 210 normal controls using the TaqMan genotyping assay. Results: We demonstrated that the ACE2 SNPs rs4646142, rs6632677, and rs2074192 were associated with COVID-19 (for all, p  <  0.05), and the differences in the ACE2 SNPs rs4646142 and rs6632677 were correlated with COVID-19-related systemic inflammatory injury and cardiovascular risk. Specifically, rs4646142 was associated with high-sensitivity C-reactive protein (hs-CRP), prealbumin (PAB), apolipoprotein A (APOA), high-density lipoprotein (HDL), and acid glycoprotein (AGP) levels. Rs6632677 was also associated with elevated CRP, acid glycoprotein (AGP), and haptoglobin (HPT). Conclusions: Our results suggest that the ACE2 SNPs rs4646142 and rs6632677 may be common genetic loci and optimal early identification genetic markers for COVID-19 with cardiovascular risk. [ABSTRACT FROM AUTHOR]

Cryptosporidium parvum (C. parvum) infection is one of the main causes of diarrhea in calves. The current study assessed the role of blood biomarkers (acute-phase proteins (APPs), procalcitonin, neopterin, cytokines, and oxidative stress in the pathogenesis, diagnosis, and prognosis of calves naturally infected with C. parvum. Fifty-seven calves, aged from 10 to 45 days, were detected positive for C. parvum and were allotted into the diseased group; twenty healthy calves were selected as a control group. Serum amyloid A, haptoglobin, cytokines, neopterin, procalcitonin, and stress biomarkers were tested in the diseased and healthy groups. The serum levels of APPs, cytokines, neopterin, procalcitonin, and malondialdehyde increased, whereas antioxidant levels were significantly decreased in diseased calves compared to the healthy group. Moreover, all examined biomarkers were significantly altered towards normal values in infected calves following different treatment protocols. All biomarkers examined were highly effective in discriminating between C. parvum-infected calves and healthy individuals. Furthermore, the area under the curve (AUC) showed that all tested parameters had a higher degree of prognostic accuracy in predicting the treatment response of calves naturally infected with C. parvum. Our data suggest the usefulness of the examined biomarkers in the immune pathogenesis of the C. parvum infection in calves, contributing to diagnosis and treatment efficacy. [ABSTRACT FROM AUTHOR]

Damage-associated molecular patterns (DAMPs) play a critical role in dendritic cells (DCs) ability to trigger a specific and efficient adaptive immune response for different physiological and pathological scenarios. We have previously identified constitutive DAMPs (HMGB1 and Calreticulin) as well as new putative inducible DAMPs such as Haptoglobin (HP), from a therapeutically used heat shock-conditioned melanoma cell lysate (called TRIMEL). Remarkably, HP was shown to be the most abundant protein in the proteomic profile of heat shock-conditioned TRIMEL samples. However, its relative contribution to the observed DCs phenotype has not been fully elucidated. Human DCs were generated from monocytes isolated from PBMC of melanoma patients and healthy donors. DC lineage was induced with rhIL-4 and rhGM-CSF. After additional stimulation with HP, the proteome of these HP-stimulated cells was characterized. In addition, DCs were phenotypically characterized by flow cytometry for canonical maturation markers and cytokine production. Finally, in vitro transmigration capacity was assessed using Transwell plates. Our results showed that the stimulation with HP was associated with the presence of exclusive and higher relative abundance of specific immune-; energy production-; lipid biosynthesis-; and DAMPs-related proteins. Importantly, HP stimulation enhanced the expression of specific DC maturation markers and pro-inflammatory and Th1-associated cytokines, and an in vitro transmigration of primary human DCs. Taken together, these data suggest that HP can be considered as a new inducible DAMP with an important role in in vitro DC activation for cancer immunotherapy. [ABSTRACT FROM AUTHOR]

Non-alcoholic fatty liver disease (NAFLD) is becoming the predominant liver disease worldwide, and vitamin E has been clinically shown to improve histological parameters in a subset of patients. In this narrative review, we investigate whether genetic factors may help to explain why some patients show histological improvements upon high-dose alpha-tocopherol (αT) treatment while others do not. In summary, we identified two factors that are associated with treatment response, including genetic variations in haptoglobin as well as fatty acid desaturase 1/2 (FADS1/FADS2). Other genetic variants such as in alpha-tocopherol transfer protein (αTTP), tocopherol associated protein (TAP), transmembrane 6 superfamily 2 (TM6SF2), cluster of differentiation 36 (CD36), and proteins involved in lipoprotein metabolism may also play a role, but have not yet been investigated in a clinical context. We propose to further validate these associations in larger populations, to then use them as a clinical tool to identify the subset of patients that will benefit the most from vitamin E supplementation. [ABSTRACT FROM AUTHOR]