Acute-phase protein concentrations in serum of clinically healthy and diseased European bison (Bison bonasus) – preliminary study

Background This is the first report describing levels of APPs in European bison. Serum concentration of acute phase proteins (APPs) may be helpful to assess general health status in wildlife and potentially useful in selecting animals for elimination. Since there is a lack of literature data regarding concentration of APPs in European bisons, establishment of the reference values is also needed. Methods A total of 87 European bison from Polish populations were divided into two groups: (1) healthy: immobilized for transportation, placing a telemetry collar and routine diagnostic purposes; and (2) selectively culled due to the poor health condition. The serum concentration of haptoglobin, serum amyloid A and α1-acid-glycoprotein were determined using commercial quantitative ELISA assays. Since none of the variables met the normality assumptions, non-parametric Mann-Whitney U test was used for all comparisons. Statistical significance was set at p < 0.05. Statistical analyses were performed using Statistica 13.3 (Tibco, USA). Results The concentration of haptoglobin and serum amyloid A was significantly higher in animals culled (euthanised) due to the poor condition in respect to the clinically healthy European bison. The levels of α1-acid-glycoprotein did not show statistical difference between healthy and sick animals. Conclusions Correlation between APPs concertation and health status was proven, therefore the determination of selected APPs may be considered in future as auxiliary predictive tool in assessing European bison health condition.

Urinary Orosomucoid (α-1-Acid Glycoprotein) As A Single Potential Biomarker Through PET Assay for Diagnosis of Early Onset Sepsis in Neonates: A Review

Neonatal sepsis is a condition in which bacteria are present in an infant’s sterile body fluids. It is considered one of the most common causes of infant death, with nearly one million deaths per birthday and approximately 2 million deaths in the first week of life. To aid in the early diagnosis of neonatal sepsis, a potential new biomarker for early neonatal sepsis called orosomucoid (ORM) or α1-glycoprotein (α1AGP) in urine is being evaluated because of its greater accuracy than current diagnostic tools. Combined with particle turbidity analysis (PET), neonatal sepsis can be diagnosed in an immediate, sensitive, specific and non-invasive manner. The early local increase in urinary ORM in sepsis suggests that it could be a new promising marker of sepsis and an important part of routine laboratory and clinical practice.

Therapeutic Effects of Mutian® Xraphconn on 141 Client-Owned Cats with Feline Infectious Peritonitis Predicted by Total Bilirubin Levels

Feline infectious peritonitis (FIP) is a fatal disease caused by feline coronavirus or its variant, referred to as the FIP virus. Recently, favorable treatment outcomes of the anti-viral drug Mutian® Xraphconn (Mutian X) were noted in cats with FIP. Thus, the therapeutic efficacy of Mutian X in cats with FIP must be explored, although the predictors of therapeutic success remain unknown. In the present study, we administered Mutian X to 141 pet cats with effusive FIP following initial veterinarian examinations. Of these, 116 cats survived but the remaining 25 died during treatment. Pre-treatment signalment, viral gene expression, and representative laboratory parameters for routine FIP diagnosis (i.e., hematocrit, albumin-to-globulin ratio, total bilirubin, serum amyloid-A, and α1-acid glycoprotein) were statistically compared between the survivor and non-survivor groups. The majority of these parameters, including hematocrit, albumin-to-globulin ratio, serum amyloid-A, α1-acid glycoprotein, and viral gene expression, were comparable between the two groups. Interestingly, however, total bilirubin levels in the survivor group were significantly lower than those in the non-survivor group (p < 0.0001). Furthermore, in almost all surviving cats with effusive FIP (96.6%, 28/29), the pre-treatment total bilirubin levels were below 0.5 mg/dL; however, the survival rate decreased drastically (14.3%, 1/7) when the pre-treatment total bilirubin levels exceeded 4.0 mg/dL. Thus, circulating total bilirubin levels may act as a prognostic risk factor for severe FIP and may serve as the predictor of the therapeutic efficacy of Mutian X against this fatal disease.

Characterisation of a new online nanoLC-CZE-MS platform and application for the glycosylation profiling of alpha-1-acid glycoprotein

The ever-increasing complexity of biological samples to be analysed by mass spectrometry has led to the necessity of sophisticated separation techniques, including multidimensional separation. Despite a high degree of orthogonality, the coupling of liquid chromatography (LC) and capillary zone electrophoresis (CZE) has not gained notable attention in research. Here, we present a heart-cut nanoLC-CZE-ESI-MS platform to analyse intact proteins. NanoLC and CZE-MS are coupled using a four-port valve with an internal nanoliter loop. NanoLC and CZE-MS conditions were optimised independently to find ideal conditions for the combined setup. The valve setup enables an ideal transfer efficiency between the dimensions while maintaining good separation conditions in both dimensions. Due to the higher loadability, the nanoLC-CZE-MS setup exhibits a 280-fold increased concentration sensitivity compared to CZE-MS. The platform was used to characterise intact human alpha-1-acid glycoprotein (AGP), an extremely heterogeneous N-glycosylated protein. With the nanoLC-CZE-MS approach, 368 glycoforms can be assigned at a concentration of 50 μg/mL as opposed to the assignment of only 186 glycoforms from 1 mg/mL by CZE-MS. Additionally, we demonstrate that glycosylation profiling is accessible for dried blood spot analysis (25 μg/mL AGP spiked), indicating the general applicability of our setup to biological matrices. The combination of high sensitivity and orthogonal selectivity in both dimensions makes the here-presented nanoLC-CZE-MS approach capable of detailed characterisation of intact proteins and their proteoforms from complex biological samples and in physiologically relevant concentrations. Graphical abstract

Profile of several autoantibodies, complement and vitamin D levels in Brazilian patients with Chikungunya infection

Introduction. Chikungunya virus infection (CKV) may lead to chronic arthritis in up to 40% of the patients. There are previous data regarding positive auto antibodies in CKV. Objective is to systematically evaluate the prevalence of auto antibodies in CKV patients. Methods. All study participants had clinical manifestations being CKV positive at least serologally or by RT-PCR data. The following autoantibodies were assessed: antinuclear antibodies (ANA), anti-dsDNA, anti-Sm, anti-Ro/SS-A, anti-La/SS-B, anti-U1RNP, IgG and IgM anticardiolipin, anticyclic citrullinated peptide (antiCCP), rheumatoid factor (RF), antiribosomal P protein, lupus anticoagulant, anti-Jo-1 and anti-Scl-70. CH100, C3 and C4 complement components, serum levels of immunoglobulins, C-reactive protein, erythrocyte sedimentation rate, alpha1-acid glycoprotein, antithyroglobulin, antithyroperoxidase, TRAb, 25 hydroxyvitamin D (25OHD), dengue and zika IgG and IgM antibodies were also measured. Results. 30 CKV patients were included. Mean age was 59.1±15.7 years, 85% females and 77% Caucasian subjects. Disease duration comprised 4.9±4.0 months. Oligoarthritis was observed in 93% cases. Mean C-reactive protein levels were 10.1±6.8 ng/dL, erythrocyte sedimentation rate — 34.2±19.9 mm/1st hour and alpha1-acid glycoprotein 115.5±52.6 mg/dL. Intramuscular betamethasone depot single dose injection was administered in 54%, prednisone — in 15% and methotrexate — in 23% cases. Importantly, 1/30 (3.3%) cases was positive for ANA, 4/30 (13.3%) — for RF and none was positive for anti-CCP or any other autoantibodies. Complement and immunoglobulin levels were all within the normal range. Low levels of 25OHD were observed in 88% cases.

Post-exercise urinary alpha-1 acid glycoprotein is not dependent on hypoxia

Introduction: Proteinuria is a transient physiologic phenomenon that occurs with a range of physical activities and during ascent to altitude. Exercise intensity appears to dictate the magnitude of post-exercise proteinuria; however, evidence also indicates possible contributions from exercise-induced hypoxemia or reoxygenation. Utilizing an environmental hypoxic chamber, this crossover designed study aimed to evaluate urinary alpha-1 acid glycoprotein (α1-AGP) excretion pre/post exercise performed in hypoxia and normoxia. Methods: Sixteen individuals underwent experimental sessions in normoxia (NOR, 20.9% O2) and hypoxia (HYP, 12.0% O2). Sessions began with a 2-hour priming period before completing a graded maximal exercise test (GXT) on a cycle ergometer, which was followed by continuation of exposure for an additional 2 hours. Physiologic responses (i.e., blood pressure, heart rate, and peripheral oxygenation), Lake Louise Scores, and urine specimens (analyzed for albumin and α1-AGP) were collected pre- and post-exercise (after 30, 60, and 120 minutes). Results: Peak power output was significantly reduced in HYP (193 ± 45 W) compared to NOR (249 ± 59 W, p < 0.01). Post-exercise urinary α1-AGP was greater in NOR (20.04 ± 14.84 μg•min-1) compared to HYP (15.08 ± 13.46 μg•min-1), albeit the difference was not significant (p > 0.05). Changes in urinary α1-AGP from pre- to post-30 minutes were not related to physiologic responses or performance outcomes observed during GXT in NOR or HYP. Conclusion: Despite profound systemic hypoxemia with maximal exercise in hypoxia, post-exercise α1-AGP excretion was not elevated above levels observed following normoxic exercise.

Serum indoxyl sulphate and its relation to albumin and α1-acid glycoprotein as a potential biomarkers of maternal intestinal metabolism during pregnancy and postpartum

Background Serum indoxyl sulfate (IS) levels depend on the production of indole in the gut. The biological effects of IS in the vascular bed could be confirmed by changes in the levels of individual serum proteins during normal pregnancy and in the postpartum period as compared with non-pregnant controls. Albumin (Alb) and α1-acid glycoprotein (AGP, orosomucoid) are the most abundant serum carrier proteins with potential interrelationships with serum levels of IS. Methods Serum levels of IS, Alb and AGP were measured in 84 pregnant women in the first, second and third trimester of pregnancy and in the postpartum period, as well as in non-pregnant controls (n = 20), using ultra-performance liquid chromatography (UPLC) coupled to mass spectrometry (IS), colorimetric assay (Alb) and immunoturbidimetric assay (AGP). Results The postpartum serum levels [mg/L] of IS were lower (p = 0.027) than in the second trimester (mean±SD: 0.85±0.39 vs 0.58±0.32). There were no differences in the IS to ALB ratio calculated in the three trimesters of pregnancy, the postpartum period, and in the non-pregnant controls. The IS/AGP ratio increased from the first to the second trimester (p = 0.039), and decreased in the postpartum period (p<0.05), when it was lower than in the second and third trimester. Conclusions The variability of the serum IS/AGP ratio during pregnancy and in the postpartum period may reflect shared involvement in the regulation of their intravascular relationships. The link between serum levels of IS derived from the gut and AGP could serve a potential biomarkers of maternal intestinal metabolism during pregnancy and postpartum.