Association between N-terminal Pro-Brain Natriuretic Peptide levels, glomerular filtration rate, and heart failure in the Multi-Ethnic Study of Atherosclerosis.

BACKGROUND: This study examined the complementary prognostic role of NT-proBNP and eGFR for predicting heart failure (HF) in adults with and without chronic kidney disease (CKD) defined as eGFR<60 ml/min/1.73m2. METHODS: We used data from the Multi-Ethnic Study of Atherosclerosis, a cohort of 6814 adults without baseline clinical cardiovascular disease. Five-year risk prediction of HF based on clinical HF risk variables (HFRV) plus NT-proBNP, eGFR or both was assessed using the C-statistic and the net reclassification index (NRI) after stratifying by CKD status. RESULTS: Mean age at baseline was 62.3±10.3 years and CKD were present in 5.9%. A total of 39 and 180 HF events occurred in participants with and without CKD, respectively. Among adults with CKD, the C-statistic for HF risk prediction increased significantly (P =0.04) from 0.71 (95% CI 0.64, 0.78) with HFRV alone to 0.78 (95% CI 0.71, 0.85) with addition of NT-proBNP. In the non-CKD group, the C-statistic increased from 0.77 (95% CI 0.74, 0.80) with HFRV alone to 0.83 (95% CI 0.80, 0.85) with addition of NT-proBNP. Further addition of eGFR to the model did not alter the C-statistic regardless of CKD status. NRI improved by 23.1% and 10.2% in CKD and non-CKD, respectively, with the addition of NT-proBNP alone and findings were similar when both eGFR and NT-proBNP were both added to model. CONCLUSIONS: In adults without clinical cardiovascular disease, the addition of NT-proBNP but not eGFR to established HFRV improves HF risk prediction in adults with and without CKD.

Postoperative Changes in the Systemic Inflammatory Milieu in Older Surgical Patients.

Dysregulated inflammation is a central component of wound healing following surgery. We prospectively enrolled older patients (n = 25, age 65 ± 7 years) undergoing elective total knee arthroplasty or total hip arthroplasty secondary to advanced osteoarthritis (OA) and healthy controls (n = 48). Inflammatory, proangiogenic (vascular endothelial growth factor [VEGF], monocyte chemoattractant protein-1 [MCP-1], and interleukin-8 [IL-8]), and antiangiogenic (interferon γ [IFN-γ] and IL-4) factors were measured using a high-sensitivity biochip. Patients with OA had significantly higher baseline VEGF (10.5 ± 1.2 pg/mL vs 4.8 ± 0.2 pg/mL, P < .001), MCP-1 (130.6 ± 7.7 pg/mL vs 88.6 ± 3.9 pg/mL, P < .0001), and IL-8 (4.0 ± 0.5 pg/mL vs 2.6 ± 0.1 pg/mL, P < .05). Postoperatively, the levels of VEGF (10.5 ± 1.2 pg/mL vs 18.8 ± 1.5 pg/mL, P < .001) and MCP-1 (130.6 ± 7.7 pg/mL vs 153.1 ± 11.5 pg/mL, P < .05) increased significantly. Baseline and postoperative MCP-1 levels correlated positively and significantly with age. The levels of IFN-γ and IL-4 (which has anti-inflammatory properties) did not significantly differ at baseline in patients with OA compared to controls and did not significantly rise postoperatively. We conclude that systemic levels of pro-inflammatory and angiogenic proteins are increased in patients with OA and rise further postoperatively, while proteins that restrain inflammation and angiogenesis do not coordinately rise. These findings implicate imbalance in inflammatory pathways in OA that may contribute to its pathobiology.

Biomarker Profile of Sepsis-Associated Coagulopathy Using Biochip Assay for Inflammatory Cytokines.

Disseminated intravascular coagulation (DIC) is a major pathophysiological mechanism of sepsis and greatly increases the risk of death in septic patients. Disseminated intravascular coagulation is a complex physiological phenomenon that involves inappropriate activation of coagulation, inflammation, and endothelial processes. The purpose of this study was to analyze the levels of inflammatory cytokines in the plasma of patients with DIC in order to compare the measured levels with those from healthy individuals, draw correlations, and provide a basis for further biomarker panel development. The inflammatory biomarkers interleukin (IL) 1ß, IL-6, IL-8, IL-10, interferon (IFN) γ, vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF) α, monocyte chemoattractant protein (MCP)-1, and epidermal growth factor (EGF) showed significant ( P < .05) elevation in patients with DIC. Interestingly, while numerous correlations were present between IL-ß, IL-6, IL-8, IL-10, IFN-γ, TNF-α, MCP-1, and many of the inflammatory cytokines measured, VEGF and EGF exhibited much less extensive correlation, suggesting that their involvement in DIC may be independent of the other investigated inflammatory markers.

Inflammatory and Metabolic Syndrome Biomarker Analysis of Vascular Outcomes in End-stage Renal Disease.

End-stage renal disease (ESRD) presents a complex syndrome in which inflammatory and metabolic processes contribute to disease progression and development of comorbid conditions. Over $1 trillion is spent globally on ESRD care. Plasma samples collected from 83 ESRD patients prior to hemodialysis were profiled for metabolic and inflammatory biomarker concentrations. Concentrations were compared between groups with and without history of stroke, acute coronary syndrome (ACS), congestive heart failure (CHF), and coronary artery disease (CAD). The 25 patients (30.1%) with history of stroke demonstrated decreased plasma interferon-γ levels (p = 0.042) and elevated plasma resistin, interleukin (IL)-1α, and leptin levels (p = 0.008, 0.021, 0.026, respectively) when compared with ESRD patients without history of stroke. The 14 patients (16.9%) with history of ACS demonstrated elevated plasma IL-6 levels (p = 0.040) when compared with ESRD patients without history of ACS. The 30 patients (36.1%) with history of CHF demonstrated decreased plasma leptin levels (p = 0.031) and elevated plasma IL-1ß levels (p = 0.042) when compared with ESRD patients without history of CHF. Finally, the 39 patients (47.0%) with history of CAD demonstrated elevated plasma IL-1α levels (p = 0.049) when compared with ESRD patients without history of CAD. Plasma biomarker concentration disturbances were observed in ESRD patients with history of stroke, ACS, CHF, and CAD when compared with ESRD patients without such history. Proinflammatory biomarker elevations were seen in stroke, ACS, CHF and CAD, while adipocytokine aberrations were observed in both stroke and CHF. These studies demonstrate that biomarker profiling of vascular comorbidities in ESRD may provide useful diagnostic and prognostic information in the management of ESRD patients.

Effect of erythropoietin stimulating agents on vascular endothelial growth factor levels in patients with end stage renal disease.

BACKGROUND: End stage renal disease (ESRD) is a debilitating disease that not only impacts quality of life, but also increases the risk of cardiovascular disease, stroke, and overall mortality. By improving our understanding of the molecular patterns seen in progression of chronic renal disease (CRD), we may be able to slow down and possibly even prevent progression renal failure. The vascular endothelial growth factor (VEGF) has been implicated as a major contributor to CRD disease progression, thus our aim was to profile the VEGF levels in patients with ESRD and to determine the effects of the erythropoietin stimulating agents (ESAs). METHODS: Under institution review board (IRB) approval, plasma samples were collected from 77 patients prior to hemodialysis and heparin administration. Normal human plasma samples (female & male, 18-35 years old) were purchased from George King Biomedical Inc (Overland Park, KS). All samples were stored at -80 °C. Inflammatory biochips were purchased from RANDOX (Co. Antrim, Northern Ireland) to test VEGF on 77 ESRD and 48 normal samples. RESULTS: The VEGF was significantly elevated in ESRD vs. normal (P<0.0001), ESRD+ESA vs. normal (P<0.0001), and ESRD-ESA vs. normal (P<0.0001). No difference was seen between ESRD+ESA and ESRD-ESA. Hemoglobin and free iron were significantly elevated in ESRD-ESA compared to ESRD+ESA (PHemoglobin=0.0007, PIron<0.0001). CONCLUSIONS: Our finding that VEGF was significantly elevated in ESRD vs. normal, aligns with the literature and suggests that VEGF may in fact play a key role in CRD progression. However, further studies to evaluate VEGF isomer levels are needed. While we saw lack of difference in VEGF levels between ESRD+ESA and ESRD-ESA, this may be due to the treatment algorithm used. Further investigation is warranted to determine whether the ESAs and hemoglobin levels show any influence on or crosstalk with the VEGF levels.

Modulation of Interleukins in Sepsis-Associated Clotting Disorders: Interplay With Hemostatic Derangement.

Interleukins play a central role in the immune system and are involved in a variety of immunological, inflammatory, and infectious disease states including sepsis syndrome. Levels of interleukins may correlate with overall survival and may directly or indirectly affect some of the regulators of coagulation and fibrinolysis, thereby disrupting hemostasis and thrombosis. Our hypothesis is that in sepsis-associated coagulopathies (SACs), interleukins may be upregulated, leading to hemostatic imbalance by generating thrombogenic mediators. We profiled the levels of interleukins IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-8, and IL-10 in addition to d-dimer (DD) in patients with SAC and in normal donors. We observed the highest increase in interleukins IL-6 (322-fold), IL-8 (48-fold), IL-10 (72-fold), and DD (18-fold). This suggests that interleukins such as IL-6 and IL-10 have a close association with coagulopathy and fibrinolytic dysregulation in sepsis and can be considered as candidates for potential therapeutic targets in SAC.

Perioperative Factors and Their Effect on the Fibrinolytic System in Arthroplasty Patients.

BACKGROUND: Total joint arthroplasty (TJA) patients are mostly of advanced age and with comorbidities such as increased body mass index (BMI) and impaired glucose tolerance. These factors and type of surgery may affect the fibrinolytic system. AIM: To investigate the effect of age, sex, BMI, type of surgery, and tranexamic acid (TXA) treatment on the fibrinolytic system in TJA patients. METHODS: Ninety-nine patients undergoing TJA (32 total hip arthroplasty [THA] and 67 total knee arthroplasty [TKA]) were included in this study. Blood samples were drawn at preoperative clinic appointments and on postoperative day 1. Antigenic levels of d-dimer, plasminogen activator inhibitor 1 (PAI-1), and tissue plasminogen activator (tPA) were measured using a commercially available enzyme-linked immunosorbent assay kit. Antiplasmin activity was measured using functional method. Age, gender, hemoglobin (Hb) levels, and BMI were collected from the records. RESULTS: Preoperative d-dimer and tPA levels were positively correlated with age, whereas preoperative antiplasmin was negatively correlated with age. Body mass index was only associated with preoperative tPA levels. There was no significant difference in postoperative levels of d-dimer, PAI-1, tPA, or antiplasmin between patients treated with TXA or without TXA. Percentage change in d-dimer and tPA showed significantly lower values in patients treated with TXA compared to the nontreated group. Type of surgery did not affect the fibrinolytic markers. CONCLUSION: These results confirm that advanced age and elevated BMI positively contribute to fibrinolytic dysregulation in TJA patients, whereas TXA seems to decrease the fibrinolytic activity.

Prevalence of metabolic syndrome in patients with end stage renal disease and relevance of biomarkers.

BACKGROUND: As chronic kidney disease (CKD) and metabolic syndrome (MetS) share many of the same risk factors and similar inflammatory pathogenesis, multiple studies have suggested a correlation between CKD and MetS.. The purpose of this study is to investigate the prevalence of MetS in end stage renal disease (ESRD) patients. Furthermore, investigating metabolic biomarker levels in patients with ESRD may provide insight into the pathogenic processes and the development of associated comorbidities. METHODS: Under IRB approval, plasma samples were collected from 89 patients with ESRD prior to hemodialysis. Biochips purchased from RANDOX (Co. Antrim, Northern Ireland) were used to test C peptide, ferritin, IL-6, resistin, insulin, TNFα, IL-1α, leptin, PAI-1 on 82 ESRD and 17 normal samples. RESULTS: All biomarkers, except insulin, were significantly elevated in patients with ESRD, suggesting an ongoing inflammatory process. Patients with ESRD+MetS, as compared to ESRD-MetS, had significantly elevated leptin. Furthermore, ESRD+MetS vs. normal was significant for leptin, but ESRD-MetS vs. normal was not. ESRD+MetS and ESRD-MetS populations were not statistically different for all other biomarkers. CONCLUSION: These findings suggest elevated Leptin in ESRD may be attributed to MetS, which is highly prevalent in the ESRD population, rather than ESRD/CKD pathogenesis alone. Lack of a significant difference for all other biomarkers suggest biomarker elevation is due to ESRD pathogenesis, rather than due to MetS as a comorbidity.

Fibrinolytic Dysregulation in Total Joint Arthroplasty Patients: Potential Clinical Implications.

BACKGROUND: The alterations of the fibrinolytic components in osteoarthritic joint disease and their postsurgical modulation are not clearly understood. Preexisting hemostatic dysfunction may lead to both thrombotic and bleeding events in these patients. AIM: To profile fibrinolytic parameters in patients undergoing total joint arthroplasty prior to and on postoperative day 1. METHODS: A total of 98 total joint arthroplasty patients were included in this study. Blood samples were drawn preoperatively and on postoperative day 1 status posttotal knee or total hip arthroplasty surgery. d-Dimer, plasminogen activator inhibitor 1 (PAI-1), and tissue plasminogen activator (tPA) were measured using commercially available enzyme-linked immunosorbent assay kits. Antiplasmin activity was measured by using a functional method. RESULTS: Preoperative PAI-1, d-dimer, and tPA levels were significantly higher in arthroplasty patients compared to healthy controls. Preoperative antiplasmin level was lower than controls. Postoperative levels of PAI-1 and d-dimer were increased compared to preoperative values. Postoperative antiplasmin values were lower than preoperative levels. Changes in tPA was not significant. There was no correlation between preoperative PAI-1 and d-dimer levels. Pre- and postoperative percentage changes in each individual were calculated for PAI-1, d-dimer, tPA, and antiplasmin. There was a positive correlation between d-dimer and PAI-1. Negative correlations between antiplasmin and d-dimer and between antiplasmin and PAI-1 were noted. CONCLUSION: These results confirm the perturbation in the fibrinolytic system of patients undergoing total joint arthroplasty surgery. Surgical intervention may also enhance the observed changes. The alterations in the fibrinolytic system may lead to the observed hemostatic complications such as bleeding, hematoma formation, or potential need for blood transfusion.

d-Dimer Levels are Significantly Increased in Blood Taken From Varicose Veins Compared With Antecubital Blood From the Same Patient.

d-Dimer is a prothrombotic biomarker and a very sensitive measure of endogenous fibrinolysis. It is used as a screening test for suspected deep vein thrombosis. This study investigated whether d-dimer levels were increased in the varicose veins of patients in comparison to their own arm samples. Patients, n = 24, 17 male, age 45 (25-91), C2-6, awaiting saphenous laser ablation were compared to matched controls, n = 24, 17 male, age 42 (24-89). Concurrent venous blood samples were taken from the arm and a lower calf/ankle (varicose) vein. The median (interquartile range) d-dimer (ng/mL) level was significantly greater in the ankle than in the arm blood of the same patient at 319 (164-631) versus 281 (167-562), P = .003, Wilcoxon. This did not occur in the controls at 269 (80-564) versus 262 (106-526), P = .361, Wilcoxon. The results indicate increased endogenous fibrinolysis in varicose veins compared with arm blood. This suggests there is more thrombotic activity or dissolution of formed subclinical fibrin thrombus which may explain the association of varicose veins with superficial vein thrombosis. This contrasts with earlier studies reporting a local reduction in fibrinolysis in venous disease.

Comparative Biochemical and Functional Studies on a Branded Human Recombinant Factor VIIa and a Biosimilar Equivalent Product.

Recombinant factor VIIa (rFVIIa; NovoSeven, Novo Nordisk, Copenhagen, Denmark) is used to control bleeding in patients with hemophilia. A generic version of FVIIa was developed by AryoGen (Tehran, Iran). This study compared the composition and functional activities of AryoSeven and NovoSeven. Each product was compared at equigravimetric (1 mg/mL) stock solution for protein content. The proteomic profile was obtained using surface-enhanced laser desorption ionization mass spectrometry. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis was carried out to determine the protein profile and Western blotting was performed using a polyclonal rabbit antihuman FVIIa antibody. The FVIIa-related antigen was also measured using a commercially available enzyme-linked immunosorbent assay method. Functional assay included the prothrombin time correction in FVII-deficient plasma. The protein content was comparable in 2 products and the mass spectra analysis showed a single peak at 50 kDa in all products. The SDS-PAGE and immunoblotting studies were comparable. Both products exhibited similar coagulant properties in different assays.

Increased expression of secreted frizzled-related protein-1 and microtubule-associated protein light chain 3 in keratoconus.

PURPOSE: To study the expression of secreted frizzled-related protein-1 (SFRP-1) and microtubule-associated protein light chain 3 (LC3), an autophagy marker, in keratoconus. METHODS: Under an institutional review board-approved protocol, de-identified and/or surgically discarded normal donor (n = 10) and keratoconus corneas (n = 10) were obtained. The corneal samples were fixed in formalin and embedded in paraffin. Immunohistochemical staining using SFRP-1 and LC3 antibodies was performed. RESULTS: The majority of expression of SFRP-1 was seen in the epithelium; however, in 3 tissues that showed high expression, staining was also present in the stroma and endothelium. Like SFRP-1, the LC3 expression in keratoconus tissues occurred at 3 different levels: low, medium, and high. Collectively these data suggest that there are differences in the expression levels of SFRP-1 and LC3 in keratoconus tissue compared with the normal tissue. Low expressivity of SFRP-1 seemed to correspond to low expressivity of LC3, whereas medium to high expressivity of SFRP-1 corresponded to medium to high expressivity of LC3. CONCLUSIONS: Increased expression of SFRP-1 and LC3 was observed in keratoconus corneas. Keratocyte autophagy seen with keratoconus may play a role in the pathogenesis of keratoconus.