Effects of anticoagulants on human plasma soluble corin levels measured by ELISA

BackgroundRecently, soluble corin was detected in human plasma. In patients with heart failure, plasma corin levels were lower than that of normal controls. In this study, we analyzed experimental conditions for measuring plasma or serum corin by an immunoassay.MethodsSerum and plasma corin levels were measured by ELISA. Effects of different anticoagulants (EDTA, heparin and sodium citrate) on plasma corin levels were examined.ResultsCorin levels in serum were similar to that in plasma with heparin (950 ± 305 vs. 929 ± 301 pg/ml, n = 40, p = 0.73), but were significantly higher than those in plasma with sodium citrate (735 ± 237 pg/ml, p < 0.01) or EDTA (716 ± 261 pg/ml, p < 0.001). Native and recombinant human corin proteins were stable in human plasma with EDTA at 4 °C or underwent freezing-and-thawing. In 348 healthy Chinese individuals, plasma corin levels ranged from 216 to 1663 pg/ml. The levels were higher in males than that in females (842 ± 283 vs. 569 ± 192 pg/ml, p < 0.001).ConclusionSoluble corin was stable in plasma samples. Plasma soluble corin levels vary depending on anticoagulants used. Samples containing heparin had significantly higher levels of corin than that in samples with EDTA or sodium citrate.     

Inhaled fentanyl citrate improves exercise endurance during high-intensity constant work rate cycle exercise in chronic obstructive pulmonary disease

ContextActivity limitation and dyspnea are the dominant symptoms of chronic obstructive pulmonary disease (COPD). Traditionally, efforts to alleviate these symptoms have focused on improving ventilatory mechanics, reducing ventilatory demand, or both of these in combination. Nevertheless, many patients with COPD remain incapacitated by dyspnea and exercise intolerance despite optimal therapy.ObjectivesTo determine the effect of single-dose inhalation of nebulized fentanyl citrate (a μ-opioid agonist drug) on exercise tolerance and dyspnea in COPD.MethodsIn a randomized, double-blind, placebo-controlled, crossover study, 12 stable patients with COPD (mean ± standard error of the mean post-β₂-agonist forced expiratory volume in one second [FEV₁] and FEV₁ to forced vital capacity ratio of 69% ± 4% predicted and 49% ± 3%, respectively) received either nebulized fentanyl citrate (50 mcg) or placebo on two separate days. After each treatment, patients performed pulmonary function tests and a symptom-limited constant work rate cycle exercise test at 75% of their maximum incremental work rate.ResultsThere were no significant postdose differences in spirometric parameters or plethysmographic lung volumes. Neither the intensity nor the unpleasantness of perceived dyspnea was, on average, significantly different at isotime (5.0 ± 0.6 minutes) or at peak exercise after treatment with fentanyl citrate vs. placebo. Compared with placebo, fentanyl citrate was associated with 1) increased exercise endurance time by 1.30 ± 0.43 minutes or 25% ± 8% (P = 0.01); 2) small but consistent increases in dynamic inspiratory capacity by ～0.10 L at isotime and at peak exercise (both P ≤ 0.03); and 3) no concomitant change in ventilatory demand, breathing pattern, pulmonary gas exchange, and/or cardiometabolic function during exercise. The mean rate of increase in dyspnea intensity (1.2 ± 0.3 vs. 2.9 ± 0.8 Borg units/minute, P = 0.03) and unpleasantness ratings (0.5 ± 0.2 vs. 2.9 ± 1.3 Borg units/minute, P = 0.06) between isotime and peak exercise was less after treatment with fentanyl citrate vs. placebo.ConclusionSingle-dose inhalation of fentanyl citrate was associated with significant and potentially clinically important improvements in exercise tolerance in COPD. These improvements were accompanied by a delay in the onset of intolerable dyspnea during exercise near the limits of tolerance.     

Plasma ascorbic acid preparation and storage for epidemiological studies using TCA precipitation

Objectives:To introduce a procedure to validate an ascorbic acid method using trichloroacetic acid (TCA) for plasma stabilization at different storage temperatures.Methods:EDTA and heparin plasma were precipitated with TCA (1:5) containing 0.54 mol/L EDTA, or without. Samples were stored at ? 20 °C and ? 70 °C and their stability was tested at room temperature for 24 h.Results:A significant 40% loss (p < 0.001) of plasma ascorbic acid was found when EDTA samples with added EDTA were stored at ? 20 °C for 2–4 weeks compared with storage at ? 70 °C. Ascorbic acid in heparin plasma without added EDTA was most unstable and samples left at room temperature for 24 h lead to almost a total loss of ascorbic acid. Addition of EDTA to the TCA solution improved stability of samples of both plasma types at room temperature.Conclusion:The recommended procedure for ascorbic acid determination in plasma stabilized with TCA is immediate storage at ? 70 °C and inclusion of EDTA into the TCA solution.     

Comparative study on the pharmacodynamics of cisatracurium: continuous infusion or intermittent bolus injection

ObjectiveTo explore a better administration way through comparison of the pharmacodynamics of cisatracurium administered by continuous infusion or intermittent bolus injection.MethodsThirty patients (ASAI-II) who had no neuromuscular disease and underwent selective surgery under general anesthesia were randomly divided into group I and II (each group with 15 patients). In group I, patients received cisatracurium by continuous infusion and in group II, by intermittent bolus injection. The responses of adductor pollicis to train-of-four (TOF) stimulation were monitored. The duration of neuromuscular blockade, recovery index and total dose of cisatracurium consumption were recorded in the two groups. Intravenous anesthesia was used for anesthesia induction and sevoflurane inhalation for maintenance of anesthesia.ResultsThe mean infusion rate was significantly lower in group I (0.78 ± 0.15 μg.kg^? 1.min^? 1) than in group II (1.09 ± 0.33 μg.kg^? 1.min^? 1) (P < 0.05). There was no significant difference in duration of neuromuscular blockade between the two groups (P > 0.05). The recovery index was 13.13 ± 3.36 min in group I and 14.38 ± 4.48 min in group II, which indicated that the recovery was faster in group I than in group II, but without statistical significance (P > 0.05). During the duration of neuromuscular blockade, 8 patients had T₁ < 3%, 4 T₁ of 3%–7% and 3 T₁ of 7%–10% in group I; T₁ was maintained between 0 and 20% in group II.ConclusionsAlthough cisatracurium consumption was significantly lower in continuous infusion than in intermittent bolus injection, continuous infusion can obtain more stable neuromuscular blockade than intermittent bolus injection.     

Anticoagulants affect matrix metalloproteinase 9 levels in blood samples of stroke patients and healthy controls

ObjectivesMatrix metalloproteinase-9 (MMP-9) represents a promising marker for acute stroke management. In clinical studies MMP-9 has been quantified by ELISA using differing protocols. We aimed to establish a valid protocol by evaluation of preanalytics.Design and methodsBlood from stroke patients (n = 28) and healthy controls (n = 28) was drawn into tubes containing different anticoagulants (EDTA, citrate, lithium-heparin (heparin) and heparin with proteinase inhibitors) and processed after 0, 60 and 240 min. MMP-9 plasma protein and mRNA from mononuclear leukocytes were determined.ResultsIn regard to anticoagulants used, samples showed different MMP-9 protein baseline values and kinetics. Stable MMP-9 protein concentrations were only measured from EDTA samples. Particularly in samples with proteinase inhibitors protein and mRNA concentrations increased over time. Kinetics did not differ between patients and controls.ConclusionsPreanalytics plays a key role for determination of MMP-9. EDTA seems to be a valid anticoagulant for MMP-9 protein measurement.     

Revaluation of biological variation of glycated hemoglobin (HbA(1c)) using an accurately designed protocol and an assay traceable to the IFCC reference system

BackgroundGlycated hemoglobin (HbA_1c) has a key role for diagnosing diabetes and monitoring glycemic state. As recently reviewed, available data on HbA_1c biological variation show marked heterogeneity. Here we experimentally revaluated these data using a well designed protocol.MethodsWe took five EDTA whole blood specimens from 18 apparently healthy subjects on the same day, every two weeks for two months. Samples were stored at ? 80 °C until analysis and assayed in duplicate in a single run by Roche Tina-quant® Gen.2 immunoassay. Data were analyzed by the ANOVA. To assess the assay traceability to the IFCC reference method, we preliminarily carried out a correlation experiment.ResultsThe bias (mean ± SD) of the Roche immunoassay was 0.3% ± 0.7%, confirming the traceability of the employed assay. No difference was found in HbA_1c values between men and women. Within- and between-subject CV were 2.5% and 7.1%, respectively. Derived desirable analytical goals for imprecision, bias, and total error resulted 1.3%, 1.9%, and 3.9%, respectively. HbA_1c had marked individuality, limiting the use of population-based reference limits for test interpretation. The estimated critical difference was ~ 10%.ConclusionsFor the first time we defined biological variation and derived indices for the clinical application of HbA_1c measurements using an accurately designed protocol and an assay standardized according to the IFCC.     

Immunosensors for quantifying cyclooxygenase 2 pain biomarkers

BackgroundCyclooxygenase 2 (COX-2) is a key enzyme in pain biomarkers, inflammation and cancer cell proliferation. Thus biosensors that can quantify pain mediators based on biochemical mechanism are imperative.MethodsBiomolecular recognition and affinity of antigenic COX-2 with the antibody were investigated using surface plasmon resonance (SPR) and ultra-sensitive portable capillary (UPAC) fluorescence sensors. Polyclonal goat anti-COX-2 (human) antibodies were covalently immobilized on gold SPR surface and direct recognition for the COX-2 antigen assessed. The UPAC sensor utilized an indirect sandwich design involving covalently attached goat anti-COX-2 as the capture antibody and rabbit anti-COX-2 (human) antibody as the secondary antibody.ResultsUPAC fluorescence signals were directly proportional to COX-2 at a linear range of 7.46 × 10^? 4–7.46 × 10¹ ng/ml with detection limit of 1.02 × 10^? 4 ng/ml. With SPR a linear range was 3.64 × 10^? 4–3.64 × 10² ng/ml was recorded and a detection limit of 1.35 × 10^? 4 ng/ml. Validation was achieved in simulated blood samples with percent recoveries of 81.39% and 87.23% for SPR and UPAC respectively.ConclusionThe developed sensors have the potential to provide objective characterization of pain biomarkers for clinical diagnoses.     

Effect of somatostatin analogue octreotide on pain relief after major abdominal surgery

BackgroundOctreotide acetate is an 8-amino-acids synthetic octapeptide analogue of somatostatin with much-enhanced duration of action and lower incidence of side effects. We assessed the utility of using intravenous octreotide as an adjuvant to opioid analgesia that might exert a post-operative opioid-sparing effect.MethodsForty-four patients were randomly allocated, to receive either a placebo or intraoperative octreotide 0.33 μg kg^?1 h^?1 intravenous infusion that was maintained in the post-operative period. Patients received for post-operative analgesia an intravenous piritramide patient controlled analgesia (PCA), set to deliver a piritramide 0.02 mg kg^?1 dose.ResultsTwo-way ANOVA revealed significantly fewer (P = 0.0003) mean ± SD weighted piritramide dose requirements in the octreotide group (19.5 ± 6.3 μg kg^?1 h^?1) than in the control group (35.7 ± 8.2 μg kg^?1 h^?1). Dunnett’s two-sided multiple-comparison post hoc test revealed a significant difference between the two groups during the first 22 post-operative hours, following which there were no differences between the two groups. There were no significant differences over time in the mean arterial pressure (P = 0.722), heart rate (P = 0.579) and respiratory rate (P = 0.823) between the octreotide group (80 ± 10 mm Hg, 74 ± 12, 14 ± 2) and the control group (82 ± 9 mm Hg, 76 ± 11, 15 ± 3), respectively.ConclusionWe demonstrated that perioperative octreotide intravenous infusion could be an adjuvant to opioid analgesia as it exerted a piritramide opioid-sparing effect. We encountered more systemic side effects such as nausea, abdominal discomfort, and diarrhea in the octreotide group than in the control group. Our findings could be beneficial to patients who cannot tolerate the adverse effects of opioids.     

Association of beta 1-adrenergic receptor gene polymorphisms with left ventricular hypertrophy in human essential hypertension

ObjectivesExperimental evidence support a key role for beta (1)-adrenergic receptor (ADRB1) in the modulation of cardiac mass. This relationship has not yet been described in Chinese population. The goal of our study was to investigate the relationships between ADRB1 gene polymorphisms and left ventricular structure in human essential hypertension.Design and methodsA total of 2417 hypertensive patients were successfully investigated. The polymorphisms of ADRB1 gene (Arg389Gly and Gly49Ser) were genotyped by using PCR-RFLP and confirmed by sequencing.ResultsPatients carrying the Arg389Arg genotype had an increase in the left ventricular septal thickness (10.4 ± 1.5 mm vs. 9.6 ± 1.5 mm, P < 0.01 or 9.4 ± 1.4 mm, P < 0.01); left ventricular posterior wall thickness (10.4 ± 2.4 mm vs. 9.6 ± 2.4 mm or 9.7 ± 2.9 mm, P < 0.01); left ventricular mass index (51.6 ± 13.3 g/m^2.7 vs. 44.6 ± 12.9 g/m^2.7, P < 0.01 or 43.2 ± 14.4 g/m^2.7, P < 0.01) and relative wall thickness (45.0 ± 9.0% vs. 42.6 ± 8.1%, P < 0.01 or 43.2 ± 8.8%, P < 0.01) as compared with those carrying genotypes Arg389Gly and Gly389Gly. These associations were independent of anthropometric factors and major clinical features and were replicated in another independent hypertensive population (n = 327).ConclusionsOur findings show that the Arg389Gly polymorphism of the ADRB1 gene confers higher risk of left ventricular hypertrophy in human essential hypertension.     

Potential negative effects of epinephrine on carotid blood flow and ETCO2 during active compression-decompression CPR utilizing an impedance threshold device

ObjectivesThis study examines the effects of IV epinephrine administration on carotid blood flow (CBF) and end tidal CO₂ (ETCO₂) production in a swine model of active compression–decompression CPR with an impedance threshold device (ACD-CPR + ITD).MethodsSix female swine (32 ± 1 kg) were anesthetized, intubated and ventilated. Intracranial, thoracic aorta and right atrial pressures were measured via indwelling catheters. CBF was recorded. ETCO₂, SpO₂ and EKG were monitored. V-fib was induced and went untreated for 6 min. Three minutes each of standard CPR (STD), STD-CPR + impedance threshold device (ITD) and active compression–decompression (ACD)-CPR + ITD were performed. At minute 9 of the resuscitation, 40 μg/kg of IV Epinephrine was administered and ACD-CPR + ITD was continued for 1 min. Statistical analysis was performed with a paired t-test. p values of <0.05 were considered statistically significant and all values are reported in mmHg unless otherwise noted.ResultsAortic pressure, cerebral and coronary perfusion pressures increased from STD < STD + ITD < ACD-CPR + ITD (p <0.001). Epinepherine administered during ACD-CPR + ITD signficantly increased mean aortic pressure (29 ± 5 vs 42 ± 12, p = 0.01), cerebral perfusion pressure (12 ± 5 vs 22 ± 10, p = 0.01), and coronary perfusion pressure (8 ± 7 vs 17 ± 4, p = 0.02); however, mean CBF and ETCO₂ decreased (respectively 29 ± 15 vs 14 ± 7.0 ml/min, p = 0.03; 20 ± 7 vs 18 ± 6, p = 0.04).ConclusionsIn this model, administration of epinepherine during ACD-CPR + ITD signficantly increased markers of macrocirculation, while significantly decreasing carotid blood flow and ETCO₂. This calls into question the ability of calculated perfusion pressures to accurately reflect oxygen delivery to end organs. The administration of epinepherine during ACD-CPR + ITD does not improve cerebral tissue perfusion.     

Comparison of normal saline, hypertonic saline and hypertonic saline colloid resuscitation fluids in an infant animal model of hypovolemic shock

PurposeIncorrect resuscitation after hypovolemic shock is a major contributor to preventable pediatric death. Several studies have demonstrated that small volumes of hypertonic or hypertonic–hyperoncotic saline can be an effective initial resuscitation solution. However, there are no pediatric studies to recommend their use. The aim of this study is to determine if in an infant animal model of hemorrhagic shock, the use of hypertonic fluids, as opposed to isotonic crystalloids, would improve global hemodynamic and perfusion parameters.MethodsExperimental, randomized animal study including thirty-four 2-to-3-month-old piglets. 30 min after controlled 30 mL kg^?1 bleed, pigs were randomized to receive either normal saline (NS) 30 mL kg^?1 (n = 11), 3% hypertonic saline (HS) 15 mL kg^?1 (n = 12), or 5% albumin plus 3% hypertonic saline (AHS) 15 mL kg^?1 (n = 11).ResultsHigh baseline heart rate (HR) and low mean arterial pressure (MAP), cardiac index (CI), brain tissue oxygenation index (bTOI), and lactate were recorded 30 min after volume withdrawal, with no significant differences between groups. Thirty minutes after volume replacement there were no significant differences between groups for HR (NS, 188 ± 14; HS, 184 ± 14; AHS, 151 ± 14 bpm); MAP (NS, 80 ± 7; HS, 86 ± 7; AHS, 87 ± 7 mmHg); CI (NS, 4.1 ± 0.4; HS, 3.9 ± 0.4; AHS, 5.1 ± 0.4 mL min^?1 m^?2); lactate (NS, 2.8 ± 0.7; HS, 2.3 ± 0.6; AHS, 2.4 ± 0.6 mmol L^?1); bTOI (NS, 43.9 ± 2.2; HS, 40.1 ± 2.5; AHS, 46.1 ± 2.3%).ConclusionsIn this model of hypovolemic shock, hypertonic fluids achieved similar end-points as twice the volume of NS. Animals treated with albumin plus hypertonic saline presented prolonged increase in blood volume parameters and recovery of the oxygen debt.     

Clinically suspected heparin-induced thrombocytopenia during extracorporeal membrane oxygenation

PurposePatients receiving extracorporeal membrane oxygenation (ECMO) are at risk for thrombocytopenia including heparin-induced thrombocytopenia (HIT). The purpose of this study was to determine the frequency of suspected HIT in patients receiving ECMO and unfractionated heparin (UFH).Materials and methodsWe conducted a retrospective review in adult patients on ECMO. Patients were included if they received ECMO for at least 5 days and concomitant UFH.ResultsThere were 119 patients who met inclusion criteria. Twenty-three patients (19%) had a heparin–platelet factor 4 immunoassay performed. Patients with suspected HIT had a significantly lower platelet count within the first 3 days of ECMO, 69 × 10⁹/L (22-126 × 10⁹/L) vs 87.5 × 10⁹/L (63-149 × 10⁹/L); P = .04. The lowest platelet count on the day of HIT testing was 43 × 10⁹/L (26-73), representing a 71% reduction from baseline. Twenty patients (87%) had an optical density score less than 0.4, and all patients had a score less than 1.0. A functional assay was performed in 7 patients (30%), with only 1 patient having laboratory-confirmed HIT.ConclusionsThe evaluation of HIT occurred in a small percentage of patients, with HIT rarely being detected. Patients who had heparin–platelet factor 4 immunoassay testing exhibited lower platelet counts with a similar duration of ECMO and UFH exposure.     

The influence of smoking on semen quality, seminal microelements and Ca2+-ATPase activity among infertile and fertile men

ObjectiveTobacco smoking is now increasing rapidly throughout the developing world and is one of the biggest threats to current and future world health. Several studies have addressed the role of cigarette smoking on semen quality, but the exact mechanisms remain inconclusive. In order to evaluate the detrimental effects of smoking on semen quality among Saudi subjects, the levels of different seminal parameters in smokers were compared to non-smokers.Patients and methodsA total of 159 semen samples (61 smokers and 98 non-smokers) from men attending an infertility clinic for routine infertility workup were sub-grouped into fertile or infertile and were compared based on standard semen analysis (according to WHO guidelines), content of metals (magnesium, zinc and cadmium) and plasma membrane Ca²⁺-ATPase activity of sperms.ResultsCadmium concentration was found significantly higher in smokers than in non-smokers either in fertile or infertile group (2.9 ± 0.4 vs 1.4 ± 0.7; 2.9 ± 0.5 vs 1.3 ± 0.7 μg L^? 1; respectively). Together with this increase in seminal Cd a significant decrease in Ca²⁺-ATPase activity (21.5 ± 2.8 vs 33.71 ± 1.2; 20.7 ± 1.5 vs 35.07 ± 2.9 mmol min^? 1 mg^? 1 protein, p < 0.05), decrease in seminal zinc (109.8 ± 8.1 vs 189.7 ± 9.9 mg L^? 1, p < 0.01) and decrease in sperm motility (41.9% ± 2.9 vs 46.01% ± 2.5; 9.8% ± 2.4 vs 15.3% ± 2.7, p < 0.05) were found.ConclusionOur data demonstrate that cigarette smoking affects both Ca²⁺-ATPase activity and motility of the spermatozoa. These effects may be attributed to increased seminal cadmium and reduced zinc concentrations.     

A feasibility study evaluating the role of cerebral oximetry in predicting return of spontaneous circulation in cardiac arrest

To date there has been no reliable noninvasive real time monitoring available to determine cerebral perfusion during cardiac arrest.ObjectivesTo investigate the feasibility of using a commercially available cerebral oximeter during in-hospital cardiac arrest, and determine whether this parameter predicts return of spontaneous circulation (ROSC).MethodsCerebral oximetry was incorporated in cardiac arrest management in 19 in-hospital cardiac arrest cases, five of whom had ROSC. The primary outcome measure was the relationship between rSO₂ and ROSC.ResultsThe use of cerebral oximetry was found to be feasible during in hospital cardiac arrest and did not interfere with management. Patients with ROSC had a significantly higher overall mean ± SE rSO₂ (35 ± 5 vs. 18 ± 0.4, p < 0.001). The difference in mean rSO₂ between survivors and non-survivors was most pronounced in the final 5 min of cardiac arrest (48 ± 1 vs. 15 ± 0.2, p < 0.0001) and appeared to herald imminent ROSC. Although spending a significantly higher portion of time with an rSO₂ > 40% was found in survivors (p < 0.0001), patients with ROSC had an rSO₂ above 30% for >50% of the duration of cardiac arrest, whereas non-survivors had an rSO₂ that was below 30% > 50% of their cardiac arrest. Patients with ROSC also had a significantly higher change in rSO₂ from baseline compared to non-survivors (310% ± 60% vs. 150% ± 27%, p < 0.05).ConclusionCerebral oximetry may have a role in predicting ROSC and the optimization of cerebral perfusion during cardiac arrest.     

Direct electrochemical determination of morphine on a novel gold nanotube arrays electrode

BackgroundMorphine (MP), a group of narcotic pain relieve drugs, is frequently used to relieve severe pain for patients, especially for those who undergo a surgical procedure. However, when overdosed or abused, MP is toxic and can cause disruption in the central nervous system. Therefore, to prevent overdose-induced toxication, it is necessary to sensitively determine the concentrations of MP in patient's blood or urine.MethodsWe report an easy-to-use approach for directly electrodepositing gold nanotube arrays onto an anodic aluminum oxide template (AAO) by performing cyclic voltammetry for a short period of time. Morphological characterization of gold nanotubes demonstrates that the inner diameter of gold nanotubes becomes smaller upon the increase of deposition time. An electrochemical sensor for direct determination of morphine was constructed by attaching gold nanotube arrays onto the surface of a glassy carbon electrode (GCE).ResultsThe resulting electrochemical sensor offers an excellent response for morphine in the concentration range of 1.22 × 10^? 7–7.44 × 10^? 4 mol/l with a detection limit of 4.06 × 10^? 8 mol/l and a correlation coefficient of 0.995.ConclusionThe electrochemical sensor shows good regeneration, stability and selectivity and has been used for the determination of morphine in serum samples with satisfied results.     

Measurement of insulin-like growth factor-1 and insulin-like growth factor binding protein-3 after delayed separation of whole blood samples

Objectives:Epidemiological studies benefit from unbiased blood specimens collected with minimal cost and effort of blood collection and storage. We evaluated the stability of IGF-1 and IGFBP-3 in whole blood samples stored at room temperature to justify delays in blood processing.Design and methods:Total IGF-1 and IGFBP-3 levels were measured in EDTA plasma (n = 12), heparin plasma (n = 12) and serum (n = 10) samples of healthy volunteers after blood processing delays up till 14 days. Stability of measured IGF-1 and IGFBP-3 levels was tested by paired t-test and a linear mixed effect model.Results:Longitudinal analysis showed that IGF-1 levels were not significantly affected by blood processing delays in EDTA tubes (p = 0.18) and IGFBP-3 levels were marginally stable (p = 0.06). In heparin plasma and serum, however, IGF-1 increased over time of delayed processing and IGFBP-3 levels tended to decrease (p < 0.01).Conclusion:Total IGF-1 and IGFBP-3 levels are stable in whole blood collected in EDTA tubes at room temperature up till 7 days, allowing a delay in blood processing to reduce costs in large multi-center studies.     

Increased serum levels of β₂-GPI-Lp(a) complexes and their association with premature atherosclerosis in patients with rheumatoid arthritis

BackgroundOur recent study found the existence of complexes of β₂-glycoprotein I (β₂-GPI) with lipoprotein(a)[Lp(a)] in circulation and the complex concentrations were increased in sera of systemic lupus erythematosus patients. The concentration of β₂-GPI-Lp(a) and its relationship with premature atherosclerosis were evaluated in rheumatoid arthritis (RA) patients.MethodsSerum concentrations of β₂-GPI-Lp(a) were measured in 53 active RA patients and 40 healthy controls by a “sandwich” ELISA. β₂-GPI-ox-LDL, ox-Lp(a), ox-LDL and anti-β₂-GPI were also measured by ELISAs. In addition, inflammatory markers were examined.ResultsSerum β₂-GPI-Lp(a) (1.12 ± 0.25 U/ml vs. 0.87 ± 0.19 U/ml, P < 0.0001) and β₂-GPI-ox-LDL (1.01 ± 0.20 U/ml vs. 0.80 ± 0.08 U/ml, P < 0.0001) concentrations in RA were both significantly higher than those of controls. Ox-Lp(a) (8.38 ± 6.69 mg/l vs. 5.49 ± 4.31 mg/l, P < 0.05) and ox-LDL (0.68 ± 0.65 mg/l vs. 0.37 ± 0.13 mg/l, P = 0.001) were also higher in RA than in controls. The area under the ROC curve (AUC) for β₂-GPI-Lp(a) (0.787) was larger than for ox-Lp(a) (0.731). AUC of β₂-GPI-ox-LDL (0.858) was also larger than for ox-LDL (0.785). β₂-GPI-Lp(a) and β₂-GPI-ox-LDL were positively correlated with ox-Lp(a), ox-LDL and CRP, respectively.Conclusionsβ₂-GPI-Lp(a) complex concentrations increased in active RA. Inflammation and oxidative stress in RA contribute to the increase of ox-Lp(a) and subsequently the formation of β₂-GPI-Lp(a).     

Impaired capsular polysaccharide is relevant to enhanced biofilm formation and lower virulence in Streptococcus pneumoniae

Streptococcus pneumoniae has been reported to form biofilms. Many different surface molecules, including capsular polysaccharide (CPS), may play a fundamental role in pneumococcal biofilm development. We designed a CPS mutant, TIGR4cps4D^?, from the TIGR4 strain and detected enhanced biofilm formation. The pathogenic diversities of the mutant were also investigated with the in vitro expression levels of pavA, lytA, IgA1, piaA, psaA, ply, and spxB. The mean OD₅₉₅ of TIGR4cps4D^? biofilm was 1.77 and 1.74, whereas that of TIGR4 was 0.76 and 0.33 on day 1 and day 2, respectively. Scanning electron microscopy and confocal laser scanning microscopy showed TIGR4cps4D^? formed a biofilm that was significantly thicker than that formed by TIGR4 (~12.22 vs. ~6.29 μm). Compared to TIGR4, the gene expression of lytA, IgA1, and, psaA in TIGR4cps4D^? was 1.9 × 10^?5-, 2.4 × 10^?5-, and 3.2 × 10^?3 fold lower under the planktonic condition, and 1.9 × 10^?5- and 9.7 × 10^?5 fold lower in biofilms, respectively. Furthermore, TIGR4cps4D^? seemed to induce less cell death, compared to the results of TIGR4 (21.38 vs. 33.47 %, after a 5-h exposure; P < 0.05). Our data indicate that impaired pneumococcal CPS may increase biofilm formation and be involved in inhibition of virulence, possibly by influencing the gene expression.     

Coenzyme Q10 levels are low and associated with increased mortality in post-cardiac arrest patients

AimSurvival after cardiac arrest (CA) is limited by the profound neurologic insult from ischemia–reperfusion injury. Therapeutic options are limited. Previous data suggest a benefit of coenzyme Q₁₀ (CoQ₁₀) in post-arrest patients. We hypothesized that plasma CoQ₁₀ levels would be low after CA and associated with poorer outcomes.MethodsProspective observational study of post-arrest patients presenting to a tertiary care center. CoQ₁₀ levels were drawn 24 h after return of spontaneous circulation (ROSC) and compared to healthy controls. Levels of inflammatory cytokines and biomarkers were analyzed. Primary endpoints were survival to discharge and neurologic status at time of discharge.Results23 CA subjects and 16 healthy controls were enrolled. CoQ₁₀ levels in CA patients (0.28 μmol L^?1, inter-quartile range (IQR): 0.22–0.39) were significantly lower than in controls (0.75 μmol L^?1, IQR: 0.61–1.08, p < 0.0001). The mean CoQ₁₀ level in CA patients who died was significantly lower than in those who survived (0.27 vs 0.47 μmol L^?1, p = 0.007). There was a significant difference in median CoQ₁₀ level between patients with a good vs poor neurological outcome (0.49 μmol L^?1, IQR: 0.30–0.67 vs 0.27 μmol L^?1, IQR: 0.21–0.30, p = 0.02). CoQ₁₀ was a statistically significant predictor of poor neurologic outcome (adjusted p = 0.02) and in-hospital mortality (adjusted p = 0.026).ConclusionCoQ₁₀ levels are low in human subjects with ROSC after cardiac arrest as compared to healthy controls. CoQ₁₀ levels were lower in those who died, as well as in those with a poor neurologic outcome.     

Inflammatory marker levels in obese adolescents with glucose intolerance: increased chitotriosidase activity

ObjectivesExistance of low grade persistent inflammation in obese children may increase the risk of metabolic and cardiovascular events. The aim was to determine whether glucose intolerance has an influence on inflammatory markers in obese adolescents.Designs and methods45 obese adolescents (mean BMI: 30.34 ± 5.42 kg/m²) were grouped as normal or impaired glucose tolerance. IL-6 and CRP levels were analyzed by commercially available kits. Chitotriosidase activity was measured by a fluorescence method and neopterin levels were determined by ELISA. Data were expressed as mean ± SD.ResultsIL-6 and CRP levels were similar in the two groups. Serum neopterin levels were not different between the groups. The chitotriosidase activity was significantly higher in the IGT group than NGT (124.33 ± 51.97 μmol/L/h vs 84.50 ± 53.99 μmol/L/h, p = 0.04).ConclusionSerum chitotriosidase activity is increased in obese adolescents with impaired glucose tolerance.