Systematic evaluation of plasma signaling cascades by functional proteomics approaches: SARS-CoV-2 infection as model.

PURPOSE: Acute phase reactants (APRs) play a critical role in inflammation. The difference in their physiological functions or the different dynamic ranges of these proteins in plasma makes it difficult to detect them simultaneously and to use several of these proteins as a tool in clinical practice. EXPERIMENTAL DESIGN: A novel multiplex assay has been designed and optimized to carry out a high-throughput and simultaneous screening of APRs, allowing the detection of each of them at the same time and in their corresponding dynamic range. RESULTS: Using Sars-CoV-2 infection as a model, it has been possible to profile different patterns of acute phase proteins that vary significantly between healthy and infected patients. In addition, severity profiles (acute respiratory distress syndrome and sepsis) have been established. CONCLUSIONS AND CLINICAL RELEVANCE: Differential profiles in acute phase proteins can serve as a diagnostic and prognostic tool, among patient stratification. The design of this new platform for their simultaneous detection paves the way for them to be more extensive use in clinical practice.

A Deep Look Into COVID-19 Severity Through Dynamic Changes in Blood Cytokine Levels.

An excessive inflammatory response to SARS-CoV-2 is thought to be a major cause of disease severity and mortality in patients with COVID-19. Longitudinal analysis of cytokine release can expand our understanding of the initial stages of disease development and help to identify early markers serving as predictors of disease severity. In this study, we performed a comprehensive analysis of 46 cytokines (including chemokines and growth factors) in the peripheral blood of a large cohort of COVID-19 patients (n=444). The patients were classified into five severity groups. Longitudinal analysis of all patients revealed two groups of cytokines, characterizing the "early" and "late" stages of the disease course and the switch between type 1 and type 2 immunity. We found significantly increased levels of cytokines associated with different severities of COVID-19, and levels of some cytokines were significantly higher during the first three days from symptom onset (DfSO) in patients who eventually required intensive care unit (ICU) therapy. Additionally, we identified nine cytokines, TNF-α, IL-10, MIG, IL-6, IP-10, M-CSF, G-CSF, GM-CSF, and IFN-α2, that can be used as good predictors of ICU requirement at 4-6 DfSO.

Acute-Phase Levels of CXCL8 as Risk Factor for Chronic Arthralgia Following Chikungunya Virus Infection.

The immunopathogenesis of chikungunya virus (CHIKV) infection and the role of acute-phase immune response on joint pain persistence is not fully understood. We investigated the profile of serum chemokine and cytokine in CHIKV-infected patients with acute disease, compared the levels of these biomarkers to those of patients with other acute febrile diseases (OAFD) and healthy controls (HC), and evaluated their role as predictors of chronic arthralgia development. Chemokines and cytokines were measured by flow Cytometric Bead Array. Patients with CHIKV infection were further categorized according to duration of arthralgia (≤ 3 months vs >3 months), presence of anti-CHIKV IgM at acute-phase sample, and number of days of symptoms at sample collection (1 vs 2-3 vs ≥4). Patients with acute CHIKV infection had significantly higher levels of CXCL8, CCL2, CXCL9, CCL5, CXCL10, IL-1ß, IL-6, IL-12, and IL-10 as compared to HC. CCL2, CCL5, and CXCL10 levels were also significantly higher in patients with CHIKV infection compared to patients with OAFD. Patients whose arthralgia lasted > 3 months had increased CXCL8 levels compared to patients whose arthralgia did not (p<0.05). Multivariable analyses further indicated that high levels of CXCL8 and female sex were associated with arthralgia lasting >3 months. Patients with chikungunya and OAFD had similar cytokine kinetics for IL-1ß, IL-12, TNF, IFN-γ, IL-2, and IL-4, although the levels were lower for CHIKV patients. This study suggests that chemokines may have an important role in the immunopathogenesis of chronic chikungunya-related arthralgia.

Soluble Transferrin Receptor during infancy and reference intervals for the Roche Cobas platform.

INTRODUCTION: Infant iron status assessments may be difficult to interpret due to infections. The soluble transferrin receptor (sTfR) has been suggested as a biomarker mainly unaffected by the acute phase response. Reference intervals reflecting dynamics of infant growth first year in life are not well established. METHODS: The sTfR and CRP concentrations were measured in samples from 451 term infants with the Roche Cobas platform in umbilical cord, at 48-96 hours, 4 and 12 months. Reference values were constructed as the 2.5th and 97.5th percentiles. The relationship between CRP concentrations >1 mg/L and sTfR was tested by Kendall correlation. RESULTS: Reference intervals for girls and boys were 2.4-9.5 mg/L at birth, 2.9-8.4 mg/L at 48-96 hours, 2.6-5.7 mg/L at 4 months and 3.0-6.3 mg/L at 12 months. No differences between sexes were observed except for at 4 months. sTfR did not covariate with CRP concentrations >1 mg/L except in 48-96 hours samples. CONCLUSION: This study reports reference intervals for sTfR from birth to 12 months of age in a large group of infants in a low-risk area for iron deficiency. sTfR might add value to infant iron status diagnostics since no covariation with CRP was found at birth, at 4 months or at 12 months.

Lipidomic analysis of local inflammation models shows a specific systemic acute phase response to lipopolysaccharides.

Toll-like receptors (TLR) are crucial for recognizing bacterial, viral or fungal pathogens and to orchestrate the appropriate immune response. The widely expressed TLR2 and TLR4 differentially recognize various pathogens to initiate partly overlapping immune cascades. To better understand the physiological consequences of both immune responses, we performed comparative lipidomic analyses of local paw inflammation in mice induced by the TLR2 and TLR4 agonists, zymosan and lipopolysaccharide (LPS), respectively, which are commonly used in models for inflammation and inflammatory pain. Doses for both agonists were chosen to cause mechanical hypersensitivity with identical strength and duration. Lipidomic analysis showed 5 h after LPS or zymosan injection in both models an increase of ether-phosphatidylcholines (PC O) and their corresponding lyso species with additional lipids being increased only in response to LPS. However, zymosan induced stronger immune cell recruitment and edema formation as compared to LPS. Importantly, only in LPS-induced inflammation the lipid profile in the contralateral paw was altered. Fittingly, the plasma level of various cytokines and chemokines, including IL-1ß and IL-6, were significantly increased only in LPS-treated mice. Accordingly LPS induced distinct changes in the lipid profiles of ipsilateral and contralateral paws. Here, oxydized fatty acids, phosphatidylcholines and phosphatidylethanolamines were uniquely upregulated on the contralateral side. Thus, both models cause increased levels of PC O and lyso-PC O lipids at the site of inflammation pointing at a common role in inflammation. Also, LPS initiates systemic changes, which can be detected by changes in the lipid profiles.

Evaluation of the level of high density lipoproteins as a predictor of severity in abdominal sepsis.

Background: Alterations in the lipid profile are part of the acute phase response, this corresponds to the so-called lipemia of sepsis. Objective: To determine if the serum level of high density lipoprotein (HDL) is related to severity and mortality. Method: Retrospective, descriptive, cross-sectional study of patients diagnosed with abdominal sepsis. During the period from April 2016 to February 2017. The severity was determined by APACHE II, SOFA, Mannheim, CONUT, the presence of organic faults and mortality. Results: We included 154 cases, 59 female and 95 male; The main organ causing abdominal sepsis was the appendix 41.6%. The overall mortality was 14.3%. The presence of organic faults was 35.1%. The mean HDL level was 37.64 mg/dl (SD ± 16.16). The findings, subjected to statistical verification by Student's t-test, showed significance among the cases with SOFA > 4 (p = 0.01) and Mannheim > 26 (p = 0.001), CONUT > 6 (p = 0.001), presence of organic failures (p = 0.001), and mortality (p = 0.003). Conclusions: HDL levels are related to severity, with the development of organic failures and mortality in sepsis.

Antecedentes: Las alteraciones en el perfil de lípidos son parte de la respuesta de fase aguda, lo que corresponde a la denominada lipemia de la sepsis. Objetivo: Determinar si la concentración sérica de lipoproteínas de alta densidad (HDL) se relaciona con la gravedad y la mortalidad. Método: Estudio retrospectivo, descriptivo, transversal, de pacientes con diagnóstico de sepsis abdominal, durante el periodo de abril de 2016 a febrero de 2017. Se determinó la gravedad mediante APACHE II, SOFA, Mannheim, CONUT, la presencia de fallas orgánicas y la mortalidad. Resultados: Se incluyeron 154 casos, 59 mujeres y 95 hombres. El principal órgano causante de sepsis abdominal fue el apéndice (41.6%). La mortalidad global fue del 14.3%. La presencia de fallas orgánicas fue del 35.1%. El valor medio de HDL se situó en 37.64 mg/dl (desviación estándar: ± 16.16). Los hallazgos, sometidos a verificación estadística mediante la prueba t de Student, mostraron significancia entre los casos con SOFA > 4 (p = 0.01) y Mannheim > 26 (p = 0.001), CONUT > 6 (p = 0.001), presencia de fallas orgánicas (p = 0.001) y mortalidad (p = 0.003). Conclusión: Los valores de HDL se relacionan con la gravedad, con el desarrollo de fallas orgánicas y con la mortalidad en la sepsis.

Vitamin D-binding protein as a biomarker to confirm specific clinical diagnoses.

Introduction: Vitamin D-binding protein (DBP) performs a variety of functions as a transporter for various ligands and takes part in a number of systemic and local physiological and pathological processes. The knowledge about the pathomechanisms of this protein involvement justifies its use as a biomarker to confirm specific clinical diagnoses suggested by nonspecific signs and symptoms.Areas covered: DBP has properties of both systemic laboratory parameters measured in the blood plasma and specific parameters measured in variety of physiological fluids to assess local changes in specific body organs. Articles published in English between 1993 and 2019 were searched for in PubMed using terms DBP, vitamin D, and metabolites, inflammation. DBP is a transport protein and a regulator of immune and inflammatory processes.Expert opinion: DBP capacity for transporting numerous ligands and co-involvement of DBP in immune and inflammatory processes suggest that DBP may be used in laboratory diagnostics as a specific parameter to confirm pathomechanisms of several systemic diseases and local conditions. Changes in the concentration of DBP present in a variety of clinical material may provide valuable information for use in assessing the severity and treatment of pathological processes.

Evaluation of commercially available three dengue rapid diagnostic test kits for diagnosis of acute dengue virus infection at the point-of-care setting in Myanmar.

Early and accurate diagnosis of dengue virus (DENV) infection is very important and Rapid Diagnostic Test (RDT) Kits are been used as a point-of-care test to check DENV infection. A Hospital and Laboratory-based descriptive study was conducted at 550-bedded Mandalay Children Hospital in 2018. Acute-phase serum samples were collected from 202 dengue suspected patients to evaluate the efficacy of RDT Kits for the diagnosis of DENV infection. Commercially available three test kits which include: ((i) CareUs Dengue Combo, Korea, (ii) Humasis Dengue Combo, Korea and (iii) Wondfo Dengue Combo, China) were validated against WHO-based reference standard tests. 140/202 patients (69.3%) was confirmed to have DENV infection. All four serotypes of dengue viruses (57 DENV-1, 7 DENV-2, 6 DENV-3 and 10 DENV-4) were identified from 80 dengue confirmed patients and DENV-1 was the dominant serotype. Combining the NS-1 antigen and IgM antibody results from the CareUs Dengue Combo Kit gave the best sensitivity (92.1%, 95% CI 86.4%-96.0%) and specificity (75.8%, 95%CI 63.3%-85.8%). Among the three RDT Kits, the performance of CareUS Kit was better than the other two. This study explored the evidence of the usefulness of RDT Kits at the point-of-care setting for diagnosis of acute dengue infection.

Evaluation of lung parenchyma, blood vessels, and peripheral blood lymphocytes as a potential source of acute phase reactants in patients with COPD.

Background: Previous studies have shown that the arterial wall is a potential source of inflammatory markers in COPD. Here, we sought to compare the expression of acute phase reactants (APRs) in COPD patients and controls both at the local (pulmonary arteries and lung parenchyma) and systemic (peripheral blood leukocytes and plasma) compartments. Methods: Consecutive patients undergoing elective surgery for suspected primary lung cancer were eligible for the study. Patients were categorized either as COPD or control group based on the spirometry results. Pulmonary arteries and lung parenchyma sections, peripheral blood leukocytes, and plasma samples were obtained from all participants. Gene expression levels of C-reactive protein (CRP) and serum amyloid A (SAA1, SAA2, and SAA4) were evaluated in tissue samples and peripheral blood leukocytes by reverse transciption-PCR. Plasma CRP and SAA protein levels were measured by enzyme-linked immunosorbent assays. Proteins were evaluated in paraffin-embedded lung tissues by immunohistochemistry. Results: A total of 40 patients with COPD and 62 controls were enrolled. We did not find significant differences in the gene expression between COPD and control group. Both CRP and SAA were overexpressed in the lung parenchyma compared with pulmonary arteries and peripheral blood leukocytes. The expression of SAA was significantly higher in the lung parenchyma than in the pulmonary artery (2-fold higher for SAA1 and SAA4, P=0.015 and P<0.001, respectively; 8-fold higher for SAA2, P<0.001) and peripheral blood leukocytes (16-fold higher for SAA1, 439-fold higher for SAA2, and 5-fold higher for SAA4; P<0.001). No correlation between plasma levels of inflammatory markers and their expression in the lung and peripheral blood leukocytes was observed. Conclusions: The expression of SAA in lung parenchyma is higher than in pulmonary artery and peripheral blood leukocytes. Notably, no associations were noted between lung expression of APRs and their circulating plasma levels, making the leakage of inflammatory proteins from the lung to the bloodstream unlikely. Based on these results, other potential sources of systemic inflammation in COPD (eg, the liver) need further scrutiny.

Acute phase response in bovine coronavirus positive post-weaned calves with diarrhea.

Bovine coronavirus (BCoV) is associated with severe diarrhea in calves, winter dysentery in adult cattle, and respiratory diseases in cattle of all ages. This study aimed to investigate the relationship between white blood cell counts and haptoglobin (Hp) and serum amyloid A (SAA) levels in post-weaned calves with diarrhea caused by BCoV and those that recovered from diarrhea. Blood and fecal samples were collected twice from the same animals; 17 post-weaned calves with diarrhea (first) and 15 post-weaned calves that recovered from diarrhea (second). Real-time polymerase chain reaction revealed that all 17 fecal samples from post-weaned calves with diarrhea and one out of 15 from diarrhea-recovered calves were positive for BCoV and negative for Cryptosporidium spp., Escherichia coli K99, Salmonella spp., bovine rotavirus, and bovine viral diarrhea virus. No Eimeria oocysts were detected using the flotation method. In comparison with post-weaned calves with diarrhea, in diarrhea-recovered calves, the lymphocyte count was significantly higher (P = 0.018), and the monocyte count was significantly lower (P = 0.001); however, the number of monocytes was still high. Post-weaned calves with diarrhea had a significantly higher Hp concentration (P < 0.001) compared with diarrhea-recovered calves. The results indicated that increased Hp concentration and monocytosis but not SAA may be associated with diarrhea caused by BCoV. The present study suggests that the monitoring of Hp concentration and monocyte count is useful in the diagnosis of post-weaned calves with diarrhea caused by BCoV in this field.

Association of acute Babesia canis infection and serum lipid, lipoprotein, and apoprotein concentrations in dogs.

BACKGROUND: Babesia canis infection induces a marked acute phase response (APR) that might be associated with alteration in lipid and lipoprotein metabolism and disease prognosis. HYPOTHESIS: Dogs with B. canis-induced APR develop dyslipidemia with altered lipoprotein concentration and morphology. ANIMALS: Twenty-nine client-owned dogs with acute B. canis infection and 10 clinically healthy control dogs. METHODS: Observational cross-sectional study. Serum amyloid A (SAA) was measured using ELISA. Cholesterol, phospholipids, and triglycerides were determined biochemically. Lipoproteins were separated using agarose gel electrophoresis. Lipoprotein diameter was assessed by polyacrylamide gradient gel electrophoresis; correlation with ApoA-1 (radioimmunoassay) and SAA was determined. RESULTS: Dogs with B. canis infection had a marked APR (median SAA, 168.3 µg/mL; range, 98.1-716.2 µg/mL) compared with controls (3.2 µg/mL, 2.0-4.2 µg/mL) (P < .001). Dogs with B. canis infection had significantly lower median cholesterol (4.79 mmol/L, 1.89-7.64 mmol/L versus 6.15 mmol/L, 4.2-7.4 mmol/L) (P = .02), phospholipid (4.64 mmol/L, 2.6-6.6 mmol/L versus 5.72 mmol/L, 4.68-7.0 mmol/L) (P = .02), and α-lipoproteins (77.5%, 27.7%-93.5% versus 89.2%, 75.1%-93.5%) (P = .04), and higher ApoA-1 (1.36 U, 0.8-2.56 U versus 0.95 U, 0.73-1.54 U) concentrations (P = .02). Serum amyloid A correlated with high-density lipoproteins (HDLs) diameter (rho = .43; P = .03) and ApoA-1 (rho = .63, P < .001). CONCLUSIONS AND CLINICAL IMPORTANCE: Major changes associated with B. canis-induced APR in dogs are related to concentration, composition, and morphology of HDL particles pointing to an altered reverse cholesterol transport. Parallel ApoA-1 and SAA concentration increase is a unique still unexplained pathophysiological finding.

Increased perioperative C-reactive protein and decreased postoperative albumin is associated with acute posttraumatic osteomyelitis in patients with high-energy tibial fractures.

BACKGROUND: Early diagnosis of acute posttraumatic osteomyelitis (POM) is of vital importance for avoiding devastating complications. Diagnosing POM is difficult due to the lack of a highly specific and sensitive test, such as in myocardial infarct, stroke and intracranial bleeding. Serum inflammatory markers, C-reactive protein (CRP), procalcitonin (PCT), white blood cells (WBC) can support clinical findings but they are not able to differentiate between inflammatory response to infection and the host response to non-infection insult with high specificity and sensitivity. AIM: The objectives of the study were to investigate whether the biochemical and immunoinflammatory patient profile could facilitate postoperative monitoring, guide the antibiotic treatment and timing of revision surgery. PATIENTS AND METHODS: This prospective nonrandomised cohort study included 86 patients after high-energy injury to the shin requiring primary surgical treatment (open or closed reduction and internal fixation of tibial fracture). Values of the biochemical and immunoinflammatory profile were measured on admission (ADD), first postoperative day (POD1) and fourth-postoperative day (POD4). RESULTS: We discovered on our sample that the development of POM is associated with increased CRP on ADD, POD1 and decreased albumins on POD4. Further studies are needed to prove that these differences can be useful in diagnosing the risk of infection. The assessment of other important risk factors such as: the extent of soft tissue damage, multiple fractures, transfusion rate, need for conversion primary external fixation to intramedullary (IM) nailing or locking plate fixation can empower our clinical judgment of POM. CONCLUSIONS: We can improve prediction of posttraumatic osteomyelitis by using the perioperative inflammatory biomarker CRP in combination with postoperative albumins levels and other associated independent risk factors.

An analysis of plasma reveals proteins in the acute phase response pathway to be candidate diagnostic biomarkers for depression.

Globally, depression is one of the most serious debilitating psychiatric mental disorders. In this study, we validated the expression levels of fibrinogen alpha (FGA), fibrinogen beta (FGB), fibrinogen gamma (FGG), Complement factor B (CFB) and serpin family D member 1(SERPIND1) in the acute phase response signaling pathway in plasma samples using enzyme-linked immunosorbent assay (ELISA).Then illuminate the roles of FGA, FGB, FGG, CFB, SERPIND1 in depression using microarray data. Gene expression dataset GSE98793 was downloaded from the Gene Expression Omnibus database. There were 128 whole blood samples included 64 patients with major depressed patients and 64 healthy controls. Differentially expressed genes (DEGs) were identified, and then protein-protein interaction (PPI) network was constructed to screen crucial genes associated with FGA, FGB, FGG, CFB and SERPIND1. Moreover, gene ontology (GO) biological processes analyses was performed. The ELISA data showed that the expression levels of FGA, FGB, FGG, CFB and SERPIND1 were up-regulated in depressed patients. The enriched GO terms were predominantly associated with the biological processes including more genes were inflammation related. The PPI network was found these five genes interacted with 11 genes. FGA, FGB, FGG, CFB and SERPIND1 may be important in the pathogenesis of depression.

Quantitative proteomics using tandem mass tags in relation to the acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin.

The inflammatory response in chickens (Gallus Gallus domesticus) is an integral part of the bird's response to infection. Detailing proteomic changes occurring during infection would be beneficial to the poultry industry, offering opportunities for comparative pathophysiological analysis. The objective of this study was to quantify the changes in the plasma proteome in chickens challenged with lipopolysaccharide (LPS), a bacterial endotoxin known to stimulate the host innate immune system. Plasma from chicken (N = 6) challenged with Escherichia coli (LPS) (2 mg/kg body weight) was collected pre (0 h) and at 12, 24, 48, and 72 h post-injection along with plasma from a control group (N = 6) challenged with sterile saline. Samples were analysed by a quantitative Tandem Mass Tags approach using a Q-Exactive-Plus mass-spectrometer. Identification and relative quantification were performed using Proteome Discoverer, and data were analysed using R. Gene Ontology terms were analysed by Cytoscape based on the Gallus gallus database. Finally, 87 significantly regulated proteins were found, including serum-amyloid-A, ovotransferrin and alpha-1-acid-glycoprotein, showing a significant effect of time post-injection in the LPS-treated group. Different pathways related with protein activation cascade and heterotopic cell-cell adhesion were affected by LPS-challenge. LPS-challenged chickens demonstrate significant changes to the plasma proteome with both increases and decreases of individual proteins within 12 h of challenge. SIGNIFICANCE: The injection of chicken with bacterial lipopolysaccharide followed by sequential plasma and clinical analysis of the bird, is a long established and a widely used model for inflammation and infection studies. This study, utilising and combining proteomic and immunoassay analysis with bioinformatic analysis, revealed that several biological pathways are modulated during this early period of inflammation. In addition, proteins with biomarker potential were identified and successfully validated. This experimental model also demonstrated potential for pathophysiological mechanism investigation and as an inflammatory model for biomedical research. There is, despite plasma being an easily accessible biological matrix which is representative of the health status of the bird, scarce data on the chicken plasma proteome. This research makes a positive contribution to the current field, generating significant data for continuing comparative analysis.

Is cord blood hepcidin influenced by the low-grade acute-phase response occurring during delivery? A small-scale longitudinal study.

AIM: To measure serum hepcidin in late pregnancy and in cord blood, and to analyze relationship between hepcidin, interleukin-6, and biomarkers of fetal iron status. MATERIALS AND METHODS: Data from 15 uncomplicated singleton pregnancies were analyzed longitudinally in trimester 3 (T3) and at birth. RESULTS: In T3, S-ferritin (median 14 µg/L) and transferrin (median 4.0 g/L) indicated low iron status, whereas the median soluble transferrin receptor (sTfR) was 4.0 mg/L, i.e. within the reference interval. Median T3 S-hepcidin was 7.8 ng/mL. Later on in cord blood, ferritin concentration (180 µg/L) were significantly higher, transferrin concentration (1.8 g/L) were significantly lower, and both sTfR (4.7 mg/L) and S-hepcidin concentrations (30.5 ng/mL) were significantly higher than maternal T3 concentrations. At the same time, cord blood interleukin-6 indicated an activated acute-phase reaction. In T3, after logarithmic transformation, there was a significant correlation between S-hepcidin and both S-ferritin (r = 0.691) and sTfR (r = -0.825). There was also a significant correlation between S-ferritin and both sTfR (r = -0.729) and transferrin (r = 0.549) in T3. CONCLUSIONS: Although S-ferritin, S-hepcidin, and sTfR were correlated during pregnancy, these relationships were not apparent in umbilical cord blood. Further, cord blood interleukin-6 indicated an activated acute-phase response, and sTfR, which is known to be unaffected by inflammation, indicated a low iron status in cord blood. Thus, instead of representing an enhanced iron status, the data appear to suggest that hepcidin and ferritin in cord blood may be influenced by the low-grade acute-phase response that occurs during delivery.

Indscators of blood lipid profile, acute phase reactions and uric acid in patients with arterial hypertension combined with connective tissue dysplasia.

OBJECTIVE: Introduction:Changes in the cardiovascular system can be divided into 2 groups due to the connective tissue dysplasia (CTD) and changes in the circulatory system, caused by pathological processes that arose on the basis of the connective structures failure. One of the risk factors of arterial hypertension (AH) remaining insufficiently studied is collagen pathology - nondifferentiated connective tissue dysplasia (NCTD). The presence of connective tissue in all organs and systems, the common origin of smooth muscles, blood and lymph from mesenchyma leads to dysplastic changes in any organ and system. NCTD is morphologically characterized by changes in collagen, elastic fibrils, glycoproteins, proteoglycans and fibroblasts, which are based on hereditary mutations of genes encoding the synthesis and spatial organization of collagen, structural proteins and protein-hydrocarbon complexes, enzymes and cofactors to them. The aim was to study external and internal phenotypic signs of CTD, indicators of blood lipid spectrum, acute phase reactions and uric acid in patients with hypertension associated with CTD. PATIENTS AND METHODS: Materials and methods: The study implied examination of 52 patients (19 women and 33 men) with AH of the 2-nd stage from 1-st to 3-rd degrees and CTD manifestations, which were on inpatient treatment in the cardiology department of the Lviv City Communal Clinical Emergency Hospital. The average age of patients was 61.14 ± 2.58 years. Patients were divided into 3 groups depending on the degree of hypertension. The first group (n = 5) included patients with AH of the 1-st degree, the second (n = 28) - with AH of the 2-nd degree, the third (n = 19) - with AH of the 3-rd degree. The control group consisted of 25 practically healthy persons. Patients underwent checkup, palpation, percussion, auscultation, laboratory examinations (blood lipid spectrum, CRP, serumukoid content and uric acid), instrumental studies (ECG, echocardiography, DBPM, ultrasound examination of the internal organs and lower limbs vessels, ultrasound examination of the sleep and vertebral arteries, X-ray examination of the bone and articular system), consultation of an ophthalmologist, a neuropathologist, a traumatologist and a dentist. RESULTS: Results: In a comparative analysis between the control group and patients with stage ІІ hypertension of 1-3 grades, in 84.6% cases external and internal CTD signs were observed, and in 15.4% cases there were no manifestations. In applying the diagnostic criteria for assessing signs of CTD and stigmata of dysebryogenesis, different numbers of points were defined depending on the severity of the AH. The highest quantity of points was found in patients of the 3-rd group (30,8 ± 0,81), which indicates a significant presence of external signs of CTD in comparison with the 1-st and 2-nd groups of patients (25.2 ± 1.38 and 26.7 ± 0.72), respectively. CONCLUSION: Conclusions: The external and internal phenotypic signs of CTD of medium and severe expressiveness degree were revealed, however, most commonly they were observed in patients with hypertension of grade 3. The presence of positive correlations between the levels of TC, HDL-C, LDL-C, AC, UA and TG indicates its direct role in the pathogenesis of hypertension, and the combination with CTD complicates the underlined pathology. Screening of the studied indicators can improve the prognosis of the course and development of cardiovascular complications.

ACUTE PHASE REACTIONS AFTER ZOLEDRONIC ACID INFUSION: PROTECTIVE ROLE OF 25-HYDROXYVITAMIN D AND PREVIOUS ORAL BISPHOSPHONATE THERAPY.

OBJECTIVE: The most common adverse reaction to zoledronic acid (ZOL) infusion is the acute phase reaction (APR), characterized by transient, usually mild, flu-like symptoms. Previous treatment with oral amino-bisphosphonates (BPs) was reported as an independent protective factor for APR, and an association between APR and 25-hydroxyvitamin D (25(OH)D) levels in BP-naïve patients treated with ZOL was identified. The aims of our study were to confirm this association and to see if it was different in patients previously treated with oral BPs compared with BP-naïve patients and to investigate the role of 25(OH)D for the time of APR onset. METHODS: We included 153 consecutive patients with postmenopausal osteoporosis undergoing their first ZOL infusion. Sixty-eight had been previously treated with oral BPs. Clinical, demographic, and serologic data were recorded. RESULTS: 25(OH)D levels were significantly lower in patients experiencing APR compared to patients without APR (26.3 ± 12.7 vs. 37.0 ± 13.5 ng/mL, respectively; P<.0001). Patients with 25(OH)D <30 ng/mL had a significantly higher risk of APR (odds ratio [OR] 4.2 [95% confidence interval [CI] 2.1-8.2]) occurring in 65%. APR was significantly less frequent in patients previously treated with oral BPs than in BP-naïve subjects (33.8% [23/68] vs 52.9% [45/85], P = .018), but only a weak association remained after correction for 25(OH)D (OR 0.5, 95% CI 0.3-1.1, P = .08). CONCLUSION: Higher baseline 25(OH)D levels appear to be protective for APR post-ZOL infusion. The role of previous treatment with oral BPs as an independent protective factor for APR should be evaluated in a larger cohort. ABBREVIATIONS: APR = acute phase reaction; BPs = amino-bisphosphonates; CI = confidence interval; 25(OH)D = 25-hydroxyvitamin D; OP = osteoporosis; OR = odds ratio; PTH = parathyroid hormone; ROC = receiver operating characteristic; ZOL = zoledronic acid.

Concentration-dependent systemic response after inhalation of nano-sized zinc oxide particles in human volunteers.

BACKGROUND: Inhalation of high concentrations of zinc oxide particles (ZnO) may cause metal fume fever. In an earlier human inhalation study, no effects were observed after exposure to ZnO concentrations of 0.5 mg/m3. Further data from experimental studies with pure ZnO in the concentration range between 0.5 and 2.5 mg/m3 are not available. It was the aim of this experimental study to establish the concentration-response relationship of pure nano-sized ZnO particles. METHODS: Sixteen healthy subjects were exposed to filtered air and ZnO particles (0.5, 1.0 and 2.0 mg/m3) for 4 h on 4 different days, including 2 h of cycling with a low workload. The effects were assessed before, immediately after, and about 24 h after each exposure. Effect parameters were symptoms, body temperature, inflammatory markers and clotting factors in blood, and lung function. RESULTS: Concentration-dependent increases in symptoms, body temperature, acute phase proteins and neutrophils in blood were detected after ZnO inhalation. Significant effects were detected with ZnO concentrations of 1.0 mg/m3 or higher, with the most sensitive parameters being inflammatory markers in blood. CONCLUSION: A concentration-response relationship with nano-sized ZnO particles in a low concentration range was demonstrated. Systemic inflammatory effects of inhaled nano-sized ZnO particles were observed at concentrations well below the occpational exposure limit for ZnO in many countries. It is recommended to reassess the exposure limit for ZnO at workplaces.

Acute-Phase Plasma Osteopontin as an Independent Predictor for Poor Outcome After Aneurysmal Subarachnoid Hemorrhage.

Experimental studies reported that osteopontin (OPN), a matricellular protein, is induced in brain after subarachnoid hemorrhage (SAH). The aim of this study was to investigate the relationships between plasma OPN levels and outcome after aneurysmal SAH in a clinical setting. This is a prospective study consisting of 109 aneurysmal SAH patients who underwent aneurysmal obliteration within 48 h of SAH. Plasma OPN concentrations were serially determined at days 1-3, 4-6, 7-9, and 10-12 after onset. Various clinical factors as well as OPN values were compared between patients with 90-day good and poor outcomes. Plasma OPN levels were significantly higher in SAH patients compared with control patients and peaked at days 4-6. Poor-outcome patients had significantly higher plasma OPN levels through all sampling points. Receiver-operating characteristic curves demonstrated that OPN levels at days 10-12 were the most useful predictor of poor outcome at cutoff values of 915.9 pmol/L (sensitivity, 0.694; specificity, 0.845). Multivariate analyses using the significant variables identified by day 3 showed that plasma OPN ≥ 955.1 pmol/L at days 1-3 (odds ratio, 10.336; 95% confidence interval, 2.563-56.077; p < 0.001) was an independent predictor of poor outcome, in addition to increasing age, preoperative World Federation of Neurological Surgeons grades IV-V, and modified Fisher grade 4. Post hoc analyses revealed no correlation between OPN levels and serum levels of C-reactive protein, a non-specific inflammatory parameter, at days 1-3. Acute-phase plasma OPN could be used as a useful prognostic biomarker in SAH.

Regulation of low-density lipoprotein cholesterol by intestinal inflammation and the acute phase response.

Systemic inflammation, induced by disease or experimental intervention, is well established to result in elevated levels of circulating triglycerides, and reduced levels of high-density lipoprotein-cholesterol (HDL-C), in most mammalian species. However, the relationship between inflammation and low-density lipoprotein-cholesterol (LDL-C) concentrations is less clear. Most reports indicate that systemic inflammation, as observed during sepsis or following high dose experimental endotoxaemia, lowers total, and LDL-C in man. However, isolated reports have suggested that certain inflammatory conditions are associated with increased LDL-C. In this review, we summarize the emerging evidence that low-grade inflammation specifically of intestinal origin may be associated with increased serum LDL-C levels. Preliminary insights into potential mechanisms that may mediate these effects, including those connecting inflammation to trans-intestinal cholesterol efflux (TICE), are considered. We conclude that this evidence supports the potential downregulation of major mediators of TICE by inflammatory mediators in vitro and during intestinal inflammation in vivo. The TICE-inflammation axis therefore merits further study in terms of its potential to regulate serum LDL-C, and as a readily druggable target for hypercholesterolaemia.