Monitoring of myocardial injury by serial measurements of QRS area and T area: The MaastrICCht cohort.

BACKGROUND: Manually derived electrocardiographic (ECG) parameters were not associated with mortality in mechanically ventilated COVID-19 patients in earlier studies, while increased high-sensitivity cardiac troponin-T (hs-cTnT) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) were. To provide evidence for vectorcardiography (VCG) measures as potential cardiac monitoring tool, we investigated VCG trajectories during critical illness. METHODS: All mechanically ventilated COVID-19 patients were included in the Maastricht Intensive Care Covid Cohort between March 2020 and October 2021. Serum hs-cTnT and NT-proBNP concentrations were measured daily. Conversion of daily 12-lead ECGs to VCGs by a MATLAB-based script provided QRS area, T area, maximal QRS amplitude, and QRS duration. Linear mixed-effect models investigated trajectories in serum and VCG markers over time between non-survivors and survivors, adjusted for confounders. RESULTS: In 322 patients, 5461 hs-cTnT, 5435 NT-proBNP concentrations and 3280 ECGs and VCGs were analyzed. Non-survivors had higher hs-cTnT concentrations at intubation and both hs-cTnT and NT-proBNP significantly increased compared with survivors. In non-survivors, the following VCG parameters decreased more when compared to survivors: QRS area (-0.27 (95% CI) (-0.37 to -0.16, p < .01) µVs per day), T area (-0.39 (-0.62 to -0.16, p < .01) µVs per day), and maximal QRS amplitude (-0.01 (-0.01 to -0.01, p < .01) mV per day). QRS duration did not differ. CONCLUSION: VCG-derived QRS area and T area decreased in non-survivors compared with survivors, suggesting that an increase in myocardial damage and tissue loss play a role in the course of critical illness and may drive mortality. These VCG markers may be used to monitor critically ill patients.

Protein S-100B, neuron-specific enolase (NSE), myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) in cerebrospinal fluid (CSF) and blood of neurological patients.

In this study, data about protein S-100B, neuron-specific enolase, myelin basic protein and glial fibrillary acidic protein in cerebrospinal fluid and blood of patients with an acute or chronic progressive neurological disorder with brain damage are reviewed. Especially in disorders with acute brain damage, determination of these proteins in CSF and blood can be helpful to establish structural and/or functional brain damage to determine severity and prognosis of the disease process and to monitor treatment effects.

Clinical validation of renin monoclonal antibody-based sandwich assays of renin and prorenin, and use of renin inhibitor to enhance prorenin immunoreactivity.

Newly developed IRMAs to measure the plasma concentrations of renin and prorenin were validated for clinical use and compared with a classical enzyme kinetic assay. The IRMAs involve two monoclonal antibodies, one that reacts equally well with renin and prorenin and one that recognizes renin well but prorenin only minimally. Prorenin reactivity with the second antibody was enhanced by adding the renin inhibitor, Remikiren, to plasma. The complex of prorenin with this active-site ligand undergoes a conformational change, whereby prorenin is converted into a form that cannot be differentiated from renin by the IRMA. The linear working range of the assay was 4.0-3000 mU/L. The concentration of prorenin was calculated by subtracting the assay result obtained without Remikiren (i.e., renin) from the result obtained with Remikiren (i.e., renin plus prorenin). No more than 2% of prorenin present in plasma was detected as renin. The interassay CVs for renin quantification were 18%, 13%, and 8% at low, medium, and high concentrations, respectively. The interassay CV for calculated prorenin was 8% at both low and high concentrations. The IRMA results were highly correlated with those of an enzyme kinetic assay in healthy subjects; in patients with such conditions as primary hyperaldosteronism, renovascular hypertension, and low-, medium-, and high-renin essential hypertension; and in women undergoing gonadotropin stimulation.

Free deoxypyridinoline in urine and serum--results in children and adolescents.

The urinary excretion of the collagen crosslinking compound dexoypyridinoline (DPD) is considered a specific index of bone resorption. Here we report on the levels of free (i.e., non peptide-bound) DPD in the urine and serum of subjects from 5 to 19 years of age, as determined by a new radioimmunoassay. Reference values for free DPD were established using 24-h urine collections from 118 healthy children and serum samples from 133 children with acute febrile illnesses. Serum and urine levels of free DPD were compared in samples from 23 short, normal children. Additionally, total (the sum of peptide-bound and free) DPD was measured by high-performance liquid chromatography in the 24-h urine collections. Urinary-free DPD was significantly correlated with total DPD (r = 0.90; P < 0.001) and declined steadily with age. Serum levels of free DPD ranged from 0.9 to 5.7 nmol/l and varied with age in boys only. No significant association was found between serum and urine levels of free DPD (r = 0.08; P = 0.37). In conclusion, urinary-free DPD did not reflect enhanced bone turnover during the time of puberty. Free DPD serum levels are very low, which may be due to rapid clearance via the kidneys.

Plasma immunoreactive endothelin-1 in pregnant women with and without pre-eclampsia.

In normotensive nonpregnant women (n = 23), the plasma concentrations of immunoreactive endothelin-1 (ir-ET-1) were not different from nonpregnant women with essential hypertension (n = 15): 3.6 (2.0-5.4) ng/L (mean, range) versus 3.8 (2.4-5.8) ng/L. In normotensive pregnant women (n = 25), the plasma level of ir-ET-1 was 2.1 (1.3-3.4) ng/L (p less than 0.01) lower than in normotensive nonpregnant women. Pre-eclamptic patients (n = 25) had elevated ir-ET-1 plasma levels of 5.0 (2.1-12.4) ng/L (p less than 0.001 for normotensive pregnant women, p less than 0.01 for nonpregnant women). The low level of ir-ET-1 in normotensive pregnant women may be explained by the increase in the distribution volume of ir-ET-1 during the course of pregnancy. Damage of vascular endothelium is a consistent morphological abnormality in pre-eclampsia. Elevated ir-ET-1 in plasma might be a biochemical marker of this abnormality and may contribute to the pathogenesis of pre-eclampsia.

Evaluation of a direct assay for atrial natriuretic peptide.

A direct radioimmunoassay for the assessment of human atrial natriuretic peptide (ANP) in plasma, using a highly specific antibody and a well-defined monoiodotyrosyl-tracer was developed and evaluated by concurrent application of an extraction method. Sensitivity was 13.4 pg/ml; intra- and interassay variations were 3.1 and 5.5%, respectively; recovery averaged 99%; normal values ranged from 15-111 pg/ml (mean +/- SD = 59 +/- 25 pg/ml, n = 41). The results, including the effect of exercise, of the two methods correlated well. Pooling ANP values in normal subjects and patients with congestive heart failure gave a good correlation (p less than 0.01). However, due to processes in unextracted plasma, in the lower range the results from the direct method were erratic. Velocity and duration of centrifugation changed the number of platelets, but no effect on ANP levels, whether assessed by the direct or by the extraction method, was observed. Although the direct method is considerably less laborious than the extracted method its lack of reliability disqualifies it for most purposes.

A sensitive radioimmunoassay of atrial natriuretic peptide in human plasma; some guidelines for clinical applications.

We report the development of an in-house RIA for human alpha-ANP. The method requires the extraction of EDTA-plasma on C18-cartridges followed by radioimmunoassay of the eluates with an antibody raised in sheep against human-alpha-ANP coupled to Keyhole Limped Hemocyanin, and a monoiodotyrosyl-alpha-ANP tracer purified with HPLC before use. The assay is precise and sensitive: intra- and interassay coefficients of variation were 8.6% and 11.6%, respectively, and the lower limit of detection was 0.8 pg/tube (4.0 pg/ml plasma). Normal values were 26.0 +/- 15.5 (mean +/- sd) pg/ml plasma (n = 25). Change from supine to sitting position significantly lowered the ANP immunoreactivity. No episodic variations were observed. Our method was applied in a study comparing plasma ANP values in samples of normal persons and CHF-patients. Assay of ANP after extraction using four different antibodies showed significant correlations. HPLC analysis of both plasma of CHF patients and plasma of normal persons revealed the existence of two major immunoreactive components besides alpha-ANP which were not detected in a radioreceptor assay.

Atrial natriuretic peptide--the influence of various physiological and sampling conditions.

A study of various physiological conditions, possibly influencing levels of atrial natriuretic peptide (ANP), is described. Atrial natriuretic peptide was determined by a radioimmunoassay, suitable for routine measurements, using 1 mL of human plasma. Atrial naturetic peptide was adsorbed onto Sep-pak C-18 cartridges, eluted with acidified ethanol and subsequently radioimmunoassayed. The detection limit was slightly less than 2 pg/tube (approximately 5 pg/mL plasma). Intra- and interassay coefficients of variation were 7 and 10%, respectively. Study of various sampling conditions revealed that blood sampling in EDTA-tubes kept on ice, and centrifuged within 1 h, gives the most reliable results. Reference values in 74 individuals ranged between 10 to 69 pg/mL (mean +/- SD = 30 +/- 11 pg/mL). We observed no difference in reference values when the blood sampling procedure was in the sitting (29 +/- 11 pg/mL, n = 43) or supine (31 +/- 12 pg/mL, n = 31) position. Venepuncture stress did not consistently change ANP-levels. No difference was observed in ANP between the follicular and the luteal phase of the menstrual cycle. ANP levels of 7 ambulant female subjects declined significantly during the study period from 7 a.m. to 3 p.m.

A sensitive radioimmunoassay of atrial natriuretic peptide in human plasma, using a tracer with an immobilized glycouril agent.

A highly specific and sensitive radioimmunoassay (RIA) for alpha-human atrial natriuretic peptide (hANP[1-28]) in plasma was developed. The assay used a [125I]monoiodotyrosyl-hANP[1-28] tracer, prepared with an immobilized glycouril agent (Protag) and purified by high pressure liquid chromatography (HPLC), and a highly specific antiserum raised against hANP[1-28], coupled to keyhole limpet haemocyanin, in sheep. Plasma was extracted using C-18 Seppak cartridges. A good parallelism was found after dilution prior to extraction of plasma of patients with congestive heart failure (CHF) or of plasma of healthy subjects. Recovery of hANP[1-28] added to plasma was 96%. The limit of detection was 0.8 pg/tube, intra- and inter-assay variation were 9 and 12%, respectively. Mean plasma ANP values in 25 normal persons with a normal salt intake was 26.0 +/- 15.5 (+/- SD) pg/ml. Plasma levels of 18 subjects (7 normals, 11 CHF) were measured using four different antisera after the extraction step. High correlations were found between the values obtained with these four antisera.