Positive Acute Phase Proteins Research Articles

Acute phase responses in clinically healthy multiparous Holsteins with and without calcium dysregulation during the early postpartum period

Patterns of calcium dysregulation resulting in low total serum calcium concentrations (tCa) at 4 DIM, known as dyscalcemia, commonly occur in multiparous Holsteins. Dyscalcemia is associated with risk of disease, decreased production, and poor reproductive performance. Inflammation is well-documented early in lactation and is associated with similarly suboptimal outcomes. The acute phase response produces markers and mediators of inflammation; therefore, the objective of our case-control study was to evaluate postpartum patterns of 3 positive acute phase proteins in cows with and without dyscalcemia. We hypothesized that dyscalcemic cows would experience more activated inflammation than eucalcemic cows and that inflammation would precede dyscalcemia diagnosis. Multiparous Holstein cows at 2 commercial dairy farms in central New York were enrolled from a parent study based on tCa at 4 DIM, at a 2:1 ratio of eucalcemic (tCa >2.3 mmol/L; n = 32) to dyscalcemic cows (tCa <2.2 mmol/L; n = 16). Blood was collected 1 to 3 d before parturition and once every 24 h postpartum through 4 DIM. Samples were analyzed for 3 acute phase proteins, serum amyloid A (SAA), haptoglobin and lipopolysaccharide binding protein (LBP). Patterns of protein concentrations in blood over time were compared using linear mixed effects models including fixed effects of calcium status group, time, parity group, farm, relevant 2-way interactions, and the random effect of cow. Overall, dynamics of acute phase proteins showed that dyscalcemic cows experienced increased acute phase responses compared with eucalcemic cows, and that these responses preceded dyscalcemia diagnosis at 4 DIM. Dyscalcemic cows had elevated concentrations of SAA beginning at 2 DIM (eucalcemic: mean = 13.88 µg/mL, 95% CI = 11.34 to 16.99 µg/mL; dyscalcemic: mean = 32.95 µg/mL, 95% CI = 24.55 to 44.21 µg/mL). and continuing through 4 DIM (eucalcemic: mean = 8.14 µg/mL, 95% CI = 6.66 to 9.95 µg/mL; dyscalcemic: mean = 30.01 µg/mL, 95% CI = 22.60 to 39.83 µg/mL). Haptoglobin concentrations were also elevated in the blood of dyscalcemic cows from 2 DIM (eucalcemic: mean = 0.39 g/L, 95% CI = 0.31 to 0.49 g/L; dyscalcemic: mean = 1.11 g/L, 95% CI = 0.79 to 1.56 g/L) through 4 DIM (eucalcemic: mean = 0.27 g/L, 95% CI = 0.21 to 0.34 g/L; dyscalcemic: mean = 1.65 g/L, 95% CI = 1.19 to 2.28 g/L). Concentrations of LBP exhibited a different pattern with a small difference between groups at 3 DIM (eucalcemic: mean = 4.67 µg/mL, 95% CI = 4.02 to 5.42 µg/mL; dyscalcemic: mean = 7.91 µg/mL, 95% CI = 6.49 to 9.63 µg/mL) that became larger at 4 DIM (eucalcemic: mean = 4.88 µg/mL, 95% CI = 4.22 to 5.64 µg/mL; dyscalcemic: mean = 10.79 µg/mL, 95% CI = 8.84 to 13.17 µg/mL). Our work supports the hypothesis that dyscalcemia and inflammatory activity are associated in dairy cows under naturally occurring postpartum conditions. While the causal structure of this relationship remains unknown, improved understanding of inflammation and dyscalcemia may provide insight into mechanisms by which some cows experience maladaptation during the early postpartum period.

The Inhibition of Serum Amyloid A Protein Aggregation by a Five-Residue Peptidomimetic: Structural and Morphological Insights.

Serum amyloid A (SAA) is a small protein consisting of 104 residues and, under physiological conditions, exists mainly in hexameric form. It belongs to the positive acute-phase proteins, which means that its plasma concentration increases rapidly in response to injury, inflammation, and infection. The accumulation of SAA molecules promotes the formation of amyloid aggregates, which deposit extracellularly in many organs, causing their dysfunction. In our previous work, we successfully designed a peptidomimetic that inhibited the aggregation of amyloidogenic SAA fragments. In the present paper, we show how the same inhibitor, named saa3Dip, affects the oligomerization and aggregation processes of MetSAA1.1 protein. The thioflavin T assay showed that saa3Dip inhibited its fibrillization. The measurement of the internal fluorophore fluorescence (Trp) showed differences that occurred in the tertiary structure of MetSAA1.1 in the presence of the inhibitor, which was also confirmed by CD spectra in the aromatic range. FTIR results suggested that saa3Dip could stabilize some fragments of the native structure of MetSAA1.1, which was confirmed by determining the melting temperature (Tm) of the protein-inhibitor complex. AFM images demonstrated that the presence of saa3Dip prevented the formation of large SAA aggregates. Our results suggest that saa3Dip stabilizes the native conformation of MetSAA1.1.

Sequence variants associated with resilient responses in growing pigs.

The current work aimed to identify genomic regions and candidate genes associated with resilience in pigs. In previous work, we proposed the body weight deviation from the expected growth curve (ΔBW) and the increase of the positive acute-phase protein haptoglobin (ΔHP) after a vaccine challenge as resilience indicators which may be improved through selective breeding in pigs. Individuals with steady growth rate and minor activation of haptoglobin (high ΔBW and low ΔHP values) were considered resilient. In contrast, pigs with perturbed growth rate and high activation of haptoglobin (low ΔBW and high ΔHP values) were considered susceptible. Both ∆BW and ∆HP were simultaneously considered to select the most resilient (N = 40) and susceptible (N = 40) pigs. A genome-wide association study was carried out for the pigs' response classification to the challenge test using whole-genome sequence data (7,760,720 variants). Eleven associated genomic regions were identified, harbouring relevant candidate genes related to the immune response (such as pro- and anti-inflammatory responses) and growth pathways. These associated genomic regions harboured 41 potential functional mutations (frameshift, splice donor, splice acceptor, start loss and stop loss/gain) in candidate genes. Overall, this study advances our knowledge about the genetic determinism of resilience, highlighting its polygenic nature and strong relationship with immunity and growth.

Longitudinal Fluctuations in Protein Concentrations and Higher-Order Structures in the Plasma Proteome of Kidney Failure Patients Subjected to a Kidney Transplant.

Using proteomics and complexome profiling, we evaluated in a year-long study longitudinal variations in the plasma proteome of kidney failure patients, prior to and after a kidney transplantation. The post-transplant period was complicated by bacterial infections, resulting in dramatic changes in the proteome, attributed to an acute phase response (APR). As positive acute phase proteins (APPs), being elevated upon inflammation, we observed the well-described C-reactive protein and Serum Amyloid A (SAA), but also Fibrinogen, Haptoglobin, Leucine-rich alpha-2-glycoprotein, Lipopolysaccharide-binding protein, Alpha-1-antitrypsin, Alpha-1-antichymotrypsin, S100, and CD14. As negative APPs, being downregulated upon inflammation, we identified the well-documented Serotransferrin and Transthyretin, but added Kallistatin, Heparin cofactor 2, and interalpha-trypsin inhibitor heavy chain H1 and H2 (ITIH1, ITIH2). For the patient with the most severe APR, we performed plasma complexome profiling by SEC-LC-MS on all longitudinal samples. We observed that several plasma proteins displaying alike concentration patterns coelute and form macromolecular complexes. By complexome profiling, we expose how SAA1 and SAA2 become incorporated into high-density lipid particles, replacing largely Apolipoprotein (APO)A1 and APOA4. Overall, our data highlight that the combination of in-depth longitudinal plasma proteome and complexome profiling can shed further light on correlated variations in the abundance of several plasma proteins upon inflammatory events.

Substance P concentration is associated with the inflammatory response and pain perception in patients with chronic pain in peripheral artery disease.

&lt;b&gt;Introduction:&lt;/b&gt; Previous studies indicate a significant role of the inflammatory response in the etiopathogenesis of peripheral artery disease (PAD) and chronic pain (CP).&lt;b&gt;Aim:&lt;/b&gt; The aim of the study was to determine the relationship between the concentration of SP and the level/concentration of inflammatory mediators (pro-inflammatory cytokines, positive and negative acute phase protein, anti-inflammatory cytokines) and pain intensity in people suffering from chronic pain (CP) in the course of PAD.&lt;b&gt;Material and methods:&lt;/b&gt; We examined 187 patients of the Department of Vascular Surgery. As many as 92 patients with PAD and CP (study group) were compared to 95 patients with PAD without CP (control group). The relationship between SP and the level/concentration of fibrinogen, C-reactive protein (CRP), antithrombin III (AT), serum albumin, interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-α) and pain intensity (Numeric Rating Scale; NRS) was analyzed. Statistical analysis was performed using the R program, assuming the level of statistical significance of α = 0.05.&lt;b&gt;Results:&lt;/b&gt; Patients with CP had significantly higher levels of fibrinogen (P &lt; 0.001), CRP (P &lt; 0.001), SP (P &lt; 0.001), IL-10 (P &lt; 0.001), and lower serum albumin levels (P &lt; 0.023). Higher SP concentration was associated with higher levels of IL-10, CRP, and pain intensity. In both groups, SP concentration correlated negatively with the level of fibrinogen (P &lt; 0.001) as well as with albumin in the control group (P &lt; 0.001).&lt;b&gt;Conclusions:&lt;/b&gt; Thus, there is a relationship between the concentration of SP and fibrinogen, along with CRP, IL-10, and the intensity of pain in people suffering from CP in the course of PAD, and the level of albumin in the group without CP.

Reticulocyte Response in Paired Blood Samples of Babesia canis Infected Young and Adult Dogs

Abstract Acute B. canis infection can lead to an acute phase reaction (APR) in dogs. The parasite invades red blood cells causing anemia through immune-mediated hemolysis and possible erythropoietic suppression. A regenerative response of the erythroid lineage during the babesiosis has not been described in extension. This research examines hematologic parameters focusing on the absolute reticulocyte count and apolipoprotein A I (ApoA I) level on the day of admission and 14 days after treatment with imidocarb-dipropionate in young (n=11) and adult (n=11) dogs naturally infected with B. canis. Metabolic and inflammatory processes were characterized by analyzing protein and lipid profiles, as well as ApoA I at specified time points. Automated analyzers were used to determine complete blood count and biochemical parameters, while ApoA I was assessed using radioimmunoassay. The reticulocyte count was determined using a manual method by means of supravital staining. Both young and adult dogs with acute B. canis infection showed non-regenerative anemia without difference. Fourteen days after successful treatment with imidocarb-dipropionate, the anemia was corrected and a high reticulocyte count was observed (p&lt;0.05). This indicates that the erythroid regenerative response was efficient in young and adult dogs, although vital signs, leukocyte count and triglyceride concentration suggest a more intense APR in young dogs. A decrease in ApoA I in both groups 14 days after treatment (p&lt;0.01) confirmed that this lipoprotein acts as a positive acute-phase protein in acute B. canis infection in dogs, but further studies are needed to connect its role in erythroid lineage regeneration.

PSVI-25 A Phytogenic Additive Improves Immune Responses of Growing Beef Cattle on Grazing System

Abstract Our objective was to evaluate the effect of a phytogenic additive in protein supplement for grazing Nellore young bulls on serum protein profile. One hundred and fourteen animals (iBW, n = 114; 273 ± 4.46 kg) were allocated for 152 days in six pastures of Brachiaria brizantha cv. Marandu (̴7 ha each), and randomly distributed in six treatments with or without additives on protein supplement (30% crude protein). Treatments were: 1) Negative Control (CON), with no additive; 2) Positive Control, with 15 ppm of Monensin (MON); 3) Digestarom (DSM Nutritional Products) at 0.2 g· animal-1· /day-1 (D0.2); 4) Digestarom at 0.4 g· animal-1· /day-1 (D0.4); 5) Digestarom at 0.6 g· animal-1· /day-1 (D0.6) and 6) Digestarom at 0.8 g· animal-1· /day-1 (D0.8). Supplement was offered ad libitum. Additives dosages were adjusted weekly to achieve the targeted dose/animal. Blood samples were collected in the last day and analyzed for positive acute phase proteins (increases with the inflammation response), haptoglobin (HAP) and glycoprotein (GLY); negative acute phase protein (decreases with the inflammation), transferrin (TRA); and Immunoglobulins A and G. Data were analyzed using SAS PROC GLIMMIX. Significance of 5% was adopted. Contrasts of use of Monensin, use of Digestarom, type of additive, and linear and quadratic effects of Digestarom were evaluated. Compared with MON, Digestarom decreased (P &amp;lt; 0.05) HAP (124 vs. 76.2 mg/dL) and GLY (1,361 vs. 85.6 mg/dL). TRA increased (P &amp;lt; 0.05) with Digestarom (310 and 307 mg/dL for D0.6 and D0.8, vs. 204 and 177 mg/dL for CON and MON). Digestarom increased (P &amp;lt; 0.05) immunoglobulins A (193, 218 and 225 mg/dL for D0.4, D0.6 and D0.8) and G (1.5 g/dL for D0.4, D0.6 and D0.8) compared with CON (IgA 146 mg/dL and IgG 0.9 g/dL) and MON (IgA 93 mg/dL and IgG 0.3 g/dL). Digestarom levels also showed a quadratic effect on TRA (including 165 and 245 mg/dL for D0.2 and D0.4) and on IgA and IgG (including 124 mg/dL and 0.8 g/dL for D0.2). Overall serum protein responses of the animals receiving Digestarom (reduced haptoglobin and glycoprotein, and graeter transferrin, IgA and IgG) demonstrates that these animals had improved immune responses and could be better able to respond to infectious and stress challenges.

Development and clinical evaluation of a quantitative fluorescent immunoassay for detecting canine CRP

ABSTRACT Canine C-reactive protein (cCRP) is one of the major positive acute phase proteins in dogs and is commonly measured to detect and monitor systemic inflammation as well as the efficacy of treatment. Traditional methods for testing cCPR, including enzyme-linked immunosorbent assay (ELISA), have some drawbacks, such as a long time for diagnosis and the requirement of well-equipped laboratories. Therefore, there is a need for a rapid and precise diagnostic test for cCRP at point-of-care. This study assessed the accuracy, precision, and validated clinical effectiveness of a diagnostic test based on fluorescent lateral flow immunoassay to detect cCRP. For the standard cCRP concentration ranging from 0 to 200 μg/mL, the cCRP diagnostic test showed strong linearity with R2 of 0.9977 (p < 0.001), and both inter- and intra-assay CVs were <14%. The limit of detection and limit of quantitation were found to be 4.0 μg/mL and 5.0 μg/mL, respectively. The cCRP serum concentration was evaluated in 21 client-owned dogs and the results were compared to a previously validated ELISA. The Pearson Correlation Coefficient between the diagnostic test kit and ELISA was 0.942 [95% confidence interval: 0.859 to 0.976, p < 0.001], and the Bland–Altman plot indicated a bias of 26.82% [95% limits of agreement: −56.03 to 109.67], indicating a significant correlation and the agreement between the data from the cCRP diagnostic test and ELISA. In conclusion, the fluorescent immunoassay based diagnostic test is a suitable option for rapidly and precisely detecting cCRP in dogs, providing a convenient alternative to traditional methods for diagnosing acute inflammation.

Divergences between serum C-reactive protein and ferritin concentrations in canine pyometra

The main aim of this report was to investigate and compare the response of serum C-reactive protein (CRP) and ferritin, two positive acute phase proteins (APPs) which usually show an increase in inflammatory processes, in dogs with pyometra. For this purpose, two different studies were made. In the first one , both proteins were measured together in an APPs profile in 25 dogs with pyometra, 25 dogs with pancreatitis (as an example of a positive inflammatory control group), and in 25 healthy dogs. In the second study, to advance the knowledge of the changes and evolution of serum ferritin and CRP in dogs with pyometra after treatment, the concentrations of both APPs were analyzed in 30 dogs with pyometra at diagnosis and after ovariohysterectomy and in 10 clinically healthy female dogs before and after elective spaying. In both studies, bitches with pyometra showed significant increases in serum CRP, indicating an inflammatory condition, but not in serum ferritin despite being a moderate positive APP. This divergence between the dynamics of these APPs could be a useful tool for the suspicion of cases of canine pyometra.

Effect of Periodontitis and Periodontal Treatment on Positive Acute Phase Proteins

Background Periodontitis is an inflammatory disease with the production of cytokines that can lead to local and systemic inflammatory responses. On the other hand, during systemic inflammation in the body, the levels of acute phase proteins (APPs) can change. APPs include positive and negative types, which in response to inflammatory conditions, the concentration of positive types increases and negative types decreases. Therefore, examining changes in APPs can be used as a marker during periodontitis and periodontal treatment. Objective The purpose of this review article was to investigate the effect of periodontitis and periodontal treatment on positive APPs. The distinguishing feature of this article is the investigation of the effect of periodontal disease and its treatment on positive APPs. In addition, the introduction of new positive APPs in the future can be used to conduct new studies. Methods In this study, the keywords periodontitis, acute phase proteins, periodontal debridement, and inflammation mediators in English were searched in Google Scholar and PubMed and Scopus databases from 2000 to 2022. Among the found articles, finally, 65 related articles were selected and reviewed. Results Periodontal treatment can reduce the levels of positive APPs increased during periodontitis. Conclusion Periodontitis as an inflammatory disease can lead to changes in the levels of positive APPs leading to an increase in the levels of these proteins. Periodontal treatment, which includes non-surgical treatment (the first step of periodontal treatment and the gold standard of this treatment is scaling above and below the gums and leveling the root surfaces to remove etiological factors) and, if necessary, surgical treatment can lead to the reduction of their levels.

The usefulness of ferritin as a biomarker in evaluating the severity of disease in wave one and wave two of COVID-19 patients – A comparative retrospective study

Background: Serum ferritin was found to be a positive acute phase protein and it was linked to a hyperinflammatory process, cytokine storm, and multiple organ damage in coronavirus disease (COVID-19). Objective: The objective of this study was to establish whether ferritin could be utilized as a biomarker to evaluate disease severity in COVID-19 waves one and two in hospitalized patients. Materials and Methods: It was an observational, retrospective study. A total of 2093 patients’ data with confirmed reverse transcriptase-polymerase chain reaction positive for COVID-19 disease were reviewed. Serum ferritin values on the day of hospital admission were compared in both waves among different grades of clinical severity correlated in terms of survival and death. Results: Survivors in wave one were 92.5% and 85.4% in wave two. In waves one and two, the nonsurvivor rate was 7.5% and 14.6%, respectively, with statistically significant P values of 0.001. Analysis of the receiver operating curve (ROC) showed the ideal ferritin cutoff point for death is 687.6 ng/mL (sensitivity: 70%; specificity: 57.3%), with an Area under curve (AUC) of 0.67, suggesting that ferritin was a better marker of mortality and severity in the first wave. In the second wave, the cutoff for severity is 583.9 ng/mL (sensitivity of 85.1%, specificity of 54.8%). AUC 0.74 suggested a marker of severity in the second wave. Conclusion: Elevated serum ferritin levels in the first wave indicated severity and mortality with exaggerated cytokine storm and early organ damage and severity in the second wave, indicating reduced inflammation and better treatment plan.

Serum amyloid A, haptoglobin and ceruloplasmin levels before and after treatment in cattle with dermatophytosis

The aim of this study is to determine the changes in serum amyloid A (SAA), haptoglobin (Hp) and ceruloplasmin (Cp) levels before and after treatment in cattle with dermatophytosis. The animal material of the study consisted of a total of 30 cattle, 20 with dermatophytosis and 10 healthy, of different ages and genders. Blood was taken from the sick animals twice, before and after the treatment, and once from the healthy animals, and their serum was separated. The obtained sera were stored at -20 ºC until the Hp, SAA and Cp levels were measured. Trichoben® (Interhas, TR) vaccine was administered to the sick animals twice, 14 days apart. When the pre-treatment and post-treatment serum Hp, SAA and Cp values of the sick animals were statistically compared, it was determined that the pre-treatment values were significantly higher (P&amp;lt;0.001) than the post-treatment values. Similarly, when the values of pre-treatment and control animals were compared, it was seen that the difference was significant (P&amp;lt;0.001). When the serum SAA, Hp and Cp values of the animals in the post-treatment and control groups were compared, it was determined that the difference was statistically insignificant (P&amp;gt;0.05). As a result, it was determined that SAA, Hp and Cp values, which are positive acute phase proteins, increased significantly in cattle with dermatophytosis and these values decreased with treatment. According to this result, it was concluded that serum SAA, Hp and Cp values are important biomarkers in the evaluation of the prognosis of the disease in cattle with dermatophytosis.

Ganoderma lucidum as Anti-Inflammatory Agent on The Level of Albumin and Globulin in Rat (Rattus Norvegicus) Rheumatoid Arthritis (RA) Model

Abstrak: Inflammation is one of the immune system's responses to infection, irritation, also cell damage. Inflammation stimulates pro-inflammatory biomarkers. Albumin and globulin are included as inflammatory biomarkers, albumin is a negative acute phase protein (-APP), which will decrease, and globulin is a positive acute phase protein (+APP), which will increase due to inflammation. Ganoderma lucidum is a medicinal mushroom with anti-inflammatory potential that could increase the albumin level in blood due to inflammation. This research aimed to examine the effect of G. lucidum extract on the albumin and globulin level of inflamed white rats anddetermine the most effective dose of extract to be an anti-inflammatory agent. The study was conducted with a completely randomized design (CRD) consisting of 6 treatments of healthy control (HC), inflamed rats as negative control (C-), inflamed rats with Na-diclofenac administration (C+), and inflamed rats with G. lucidum extract administration with a dose of 250, 500, 750 mg/kg BW (T1, T2, and T3) with four replications each. The independent variable is the dose variant of G. lucidum (250, 500, 750 mg/kg BW), with the dependent variable being the changesin albumin and globulin levels. The main parameters are albumin and globulin levels, and the support parameter is the GC-MS test. The data were processed using Analysis of Variance (ANOVA) at an error rate of 5%, followed by Duncan's analysis at 95% confidence level. The results show that mushroom G. lucidum extract administration with a dose of 250 mg/kg BW is the most effective dose to be ana dose of 250 mg/kg BW, which is the most effective anti-inflammatory agent.

The biological role of IL-1, IL-6 and CRP and their application in the diagnosis of the inflammatory process

Activation of inflammation is a type of innate, non-specific defence of the body against harmful factors of external or internal origin. During this process, various types of cytokines are released, including interleukin-1 (IL-1) and interleukin-6 (IL-6), responsible for the intensification of inflammatory reactions and the activation of hepatic acute phase proteins synthesis. IL-1 exists in two isoforms (IL-1α and IL-1β), showing similar, pro-inflammatory biological properties, but differing in origin, place of release and method of activation. The use of IL-1 in the diagnosis of the inflammatory process is limited, mainly due to the short half-life of this cytokine. IL-6 has a pleiotropic nature of action by using different types of receptors signaling. IL-6 is responsible for activating the synthesis of a wide range of acute phase proteins, also is involved in hematopoiesis and the immune response. IL-6 is a good biomarker of the early phase of inflammation because it has a longer half-life than other cytokines, and its concentration in the blood may increase several thousand times during the first hours after the initiation of inflammation. Due to its high sensitivity in detecting inflammation, C-reactive protein is the most commonly determined highly positive acute phase protein. During the inflammatory process, the pentameric CRP isoform (pCRP) dissociates into monomeric CRP (mCRP), which changes the anti-inflammatory properties of CRP into highly pro-inflammatory. The currently used laboratory tests detect only the pCRP isoform, because it is well soluble in blood and accumulates in it when conversion to mCRP is no longer effective.

High-throughput immunoaffinity enrichment and N-glycan analysis of human plasma haptoglobin.

Haptoglobin (Hp) is a positive acute phase protein, synthesized in the liver, with four N-glycosylation sites carrying mainly complex type N-glycans. Its glycosylation is altered in different types of diseases but still has not been extensively studied mainly due to analytical challenges, especially the lack of a fast, efficient, and robust high-throughput Hp isolation procedure. Here, we describe the development of a high-throughput method for Hp enrichment from human plasma, based on monolithic chromatographic support in immunoaffinity mode and downstream Hp N-glycome analysis by hydrophilic interaction ultrahigh-performance liquid chromatography with fluorescent detection (HILIC-UHPLC-FLR). Chromatographic monolithic supports in a 96-well format enable fast, efficient, and robust Hp enrichment directly from diluted plasma samples. The N-glycome analysis demonstrated that a degree of Hp deglycosylation differs depending on the conditions used for N-glycan release and on the specific glycosylation site, with Asn 241 being the most resistant to deglycosylation under tested conditions. HILIC-UHPLC-FLR analysis enables robust quantification of 28 individual chromatographic peaks, in which N-glycan compositions were determined by UHPLC coupled to electrospray ionization quadrupole time of flight mass spectrometry. The developed analytical approach enables fast evaluation of total Hp N-glycosylation and is applicable in large-scale studies.

Plasma bupivacaine levels (total and free/unbound) during epidural infusion in liver resection patients: a prospective, observational study

IntroductionLiver resection patients may be at an increased risk of local anesthetic (LA) toxicity because the liver is essential for metabolizing LA and producing proteins (mainly α1-acid glycoprotein (AAG)) that...

Inflammation and Loss of Skeletal Muscle Mass in Chronic Limb Threatening Ischemia

Lower extremity peripheral arterial disease (PAD) is an atherosclerotic disease of the lower extremities. Atherosclerosis, inflammation, and sarcopenia are independently associated and potentiate each other. Inflammation is deeply involved in the formation and progression of atherosclerosis and is also involved in the pathophysiology of sarcopenia. Sarcopenia is defined as low muscle mass, with low muscle strength. This study aims to determine the differences in skeletal muscle characteristics and in inflammatory parameters between patients with claudication and with chronic limb threatening ischemia (CLTI). An observational, prospective study in patients with PAD was conducted from January 2018 to December 2020. The clinical characteristics and the cardiovascular risk factors were prospectively registered. The inflammatory parameters determined were: positive acute phase proteins (C-reactive Protein- CRP- and fibrinogen) and negative acute phase proteins albumin, total cholesterol and high-density lipoprotein (HDL). The skeletal muscle area and density were quantified with a computed topography (CT) scan. The strength was determined with a Jamar® hydraulic hand dynamometer. A total of 116 patients (mean age: 67.65±9.53 years-old) 64% with claudication and 46% with CLTI were enrolled in the study. No differences were registered between patients with claudication and CLTI on age, cardiovascular risk factors (hypertension, dyslipidemia, diabetes mellitus, and smoking habits) and medication. There was a higher prevalence of men in the claudication group (88.89% vs. 71.70%, P=0.019). Analyzing the inflammatory parameters, we noted that patients with CLTI had increased serum levels of positive acute phase proteins: CRP (37.53±46.61mg/L vs. 9.18±26.12mg/L, P=0.000), and fibrinogen (466.18±208.07mg/dL vs. 317.37±79.42mg/dL, P=0.000). CLTI patients had decreased negative acute phase proteins: albumin (3.53±0.85g/dL vs. 3.91±0.72g/dL, P=0.001), total cholesterol (145.41±38.59mg/dL vs. 161.84±34.94mg/dL, P=0.013) and HDL (38.70±12.19mg/dL vs. 51.31±15.85mg/dL, P=0.000). We noted that patients with CLTIhad lower skeletal muscle area and mass (14,349.77±3,036.60mm2 vs. 15,690.56±3,183.97mm2P=0.013; 10.11±17.03HU vs. 18.02±13.63HU P=0.013). After adjusting for the variable sex, the association between skeletal muscle density and CLTI persisted (r (97)=-0.232, P=0.021). The groups did not differ in strength (patients with claudication: 25.39±8.23 Kgf vs. CLTI: 25.17±11.95 Kgf P=0.910). CLTI patients have decreased skeletal muscle mass and a systemic inflammation status. Recognizing the deleterious triad of atherosclerosis, inflammation and loss of skeletal mass patients with CLTI is an opportunity to improve medical therapy and to perform a timely intervention to stop this vicious cycle.

Fatigue and Pain Severity in Gulf War Illness Is Associated With Changes in Inflammatory Cytokines and Positive Acute Phase Proteins.

The aim of the study is to investigate relationships between inflammatory analytes and symptoms of pain and fatigue in Gulf War illness (GWI). In this preliminary study, 12 male veterans meeting GWI criteria provided daily blood samples and symptom ratings over 25 days. Linear mixed models were used to analyze associations between symptoms and sera concentrations of cytokines, acute phase proteins, insulin, and brain-derived neurotropic factor. Analyses included 277 days with both blood draws and self-reports. Days with worse fatigue severity were associated with higher C-reactive protein and serum amyloid A, and lower eotaxin 1. Muscle pain and joint pain were associated with leptin, monocyte chemoattractant protein 1, and interferon γ-induced protein. Joint pain was further associated with serum amyloid A and eotaxin 3. Gulf War illness involves fatigue and pain associated with inflammation. Conventional and novel anti-inflammatories should be further explored for the treatment of GWI.

Evaluation of acute phase proteins, adiponectin and endothelin-1 to determine vascular damage in dogs with heartworm disease (Dirofilaria immitis), before and after adulticide treatment

Previous studies have shown the existence of an acute phase response in dogs with heartworm (Dirofilaria immitis), probably caused by the vascular inflammation that occurs during the pathogenesis of this disease. In addition, it has been seen that this acute phase response persists after finishing treatment, especially in dogs with pulmonary hypertension (PH). Furthermore, echocardiographic studies have shown that PH and endarteritis appear to persist for at least 10 months after completion of adulticide treatment, suggesting that the vascular changes in these dogs may not be reversible. Therefore, the objective of this study was to evaluate the serum concentrations of different positive acute phase proteins (APP) [C reactive-protein (CRP), haptoglobin and ferritin] and negative APP (albumin and paraoxonase-1 (PON-1)), and the usefulness of the endothelin-1 (ET-1) and adiponectin, in dogs infected by D. immitis to evaluate their usefulness as diagnostic biomarkers of vascular damage and PH and their progression throughout therapy up to 7 months after the end of adulticide treatment. Twenty-five heartworm-infected dogs received adulticide treatment, and serum measurements were performed on the day of diagnosis (day 0), day of discharge (day 90), and 6 months after discharge (day 270). In addition, presence or absence of PH was also echocardiographically determined using the Right Pulmonary Artery Distensibility Index. PH was present in 44% of the dogs on day 0 and day 90, and in 48% of dogs on day 270. Alterations were observed in the concentrations of all APP throughout the study, persisting the alterations in PON-1 and ferritin on day 270. Depending on the presence or absence of PH, CRP showed significant differences throughout the study, as did ET-1. On the other hand, adiponectin did not show variations throughout the study, so it did not seem a useful marker in this disease. These results could reflect the possible persistence of vascular inflammation up to 7 months after finishing treatment, whether or not there was PH, and consolidate the study of APP as useful markers in heartworm disease. Moreover, persistent PH could be the consequent clinical manifestation in dogs with more severe vascular alterations so the study of APP, especially CRP, and ET-1 could be especially advantageous in these patients in the early evaluation of the disease, as well as for the determination of disease severity, monitoring therapeutic responses, and predicting outcomes.

Orosomucoid: a promising biomarker for the assessment of exercise-induced fatigue triggered by basic combat training

BackgroundOrosomucoid (ORM) is a positive acute phase protein verified to be upregulated in various forms of exercise-induced fatigued (EIF) rodents. However, its association with EIF among human beings remained unknown. This study aimed to explore the association between serum ORM and EIF triggered by military basic combat training (BCT).MethodsThe degree of EIF were measured by Borg’s Rating of Perceived Exertion Scale (Borg-RPE-Scale®) as RPE score after BCT. Fifty-three male recruits were classified into three groups according to the RPE score: (1) group 1 (slight fatigue group): RPE score after BCT < 13; (2) group 2 (moderate fatigue group): RPE score after BCT = 13 or 14; (3) group 3 (severe fatigue group): RPE score after BCT > 14. The levels of blood ORM, lactate (LAC), cortisol and C-reactive protein (CRP) were determined before and after BCT. The diagnostic value of ORM was evaluated by receiver operating characteristic (ROC) curve analysis and logistic regression.ResultsAfter BCT, the level of LAC, CRP, and cortisol increased among all groups, but the changes had no significant between-group difference (all p > 0.05). The level of ORM had a specific significant increase in group 3 (p = 0.039), and the changes of ORM (ΔORM) had significant difference among groups (p = 0.033). ROC curve analysis showed that the estimated area under ROC curve for ΔORM was 0.724 (p = 0.009) with the recommended optimal cut-off value as 0.2565 mg/mL. Logistic analysis showed that recruits with ΔORM ≥ 0.2565 mg/mL had higher odds for suffering from severe EIF, 5.625 times (95% CI 1.542–20.523, p = 0.009) as large as those with ΔORM < 0.2565 mg/mL.ConclusionORM might be a promising biomarker of severe EIF triggered by BCT among male recruits. Its potential optimal cut-off value regarding ΔORM was recommended to be 0.2565 mg/mL.