ACTH, Cortisol and IL-6 Levels in Athletes following Magnesium Supplementation.

BACKGROUND: Physical exercise activates the hypothalamo-pituitary-adrenal (HPA) axis and induces the body's inflammatory response. Due to contemporary dietary habits and increased energy expenditure, athletes are susceptible to depletion of magnesium ions. The aim of our study was to investigate, through assessment of plasma ACTH, serum IL-6, and salivary/serum cortisol levels, if chronic magnesium supplementation might reduce damaging stress effects in amateur rugby players. METHODS: Rugby players (N=23) were randomly assigned to intervention and control group. Basal samples were collected before intervention group started a 4-week-long supplementation with magnesium (500 mg Mg/d). Blood and saliva sampling were done a day before the match (Day-1), on the morning of competition (Game), and during a six-day-long recovery period (Day1, Day3 and Day6). ACTH, serum/salivary cortisol, IL-6 and total/differential leukocytes counts were determined at each time point. RESULTS: There was a statistically significant increase in ACTH concentration in intervention group compared to control group, while reductions in cortisol concentrations between the two groups were the greatest at Day-1 (p < 0.01) and at the day of competition (Game) (p < 0.01). Our results revealed that magnesium completely abolished the increase in IL-6 level noted in control group on Day1 and Day3 vs. Day-1 (p < 0.01) and also diminished the rise in neutrophil/lymphocyte ratio in intervention group vs. control group (p < 0.01). CONCLUSIONS: These results suggest the possibly important influence magnesium supplementation might have on the change of parameters of HPA axis activity and reduction of immune response activation following strenuous physical exercise such as a rugby game.

Cystatin C and serum creatinine as predictors of kidney graft outcome.

PURPOSE: Serum cystatin C (Cys C) was evaluated as a predictor of kidney graft failure progression, and its predictive ability was compared to other markers of graft function. METHODS: The following kidney graft markers were determined in 91 patients who came for regular checkups of kidney graft function to our outpatient service in February 2008: Cys C, serum creatinine (sCr), 24-h proteinuria and 24-h urinary creatinine clearance (CCr). Glomerular filtration rate (eGFR) was estimated using sCr-based and Cys C formula. Patients were regularly monitored until February 2013 or to graft failure. RESULTS: During follow-up, graft failure occurred in 21 recipients. The Cys C ≥2.65 mg/l discriminated patients with and without graft failure (sensitivity of 80.95% and specificity of 92.86%). According to c statistic, the highest performance was achieved for Cys C (0.874). In addition, Cys C area under the curve (AUC) was significantly better than CCr AUC (p = 0.007), 24-h proteinuria AUC (p = 0.03), eGFR estimated by the chronic kidney disease epidemiology collaboration (EPI) AUC (p = 0.05), but not better than sCr or eGFR AUCs calculated by other formulas. In the multivariable model, sCr, CCr, Cys C and eGFRs were predictors of graft failure. Combination of Cys C, sCr and logarithm of 24 h proteinuria (0.883) or Cys C, CCr and logarithm of 24-h proteinuria (0.884) had the highest AUC for predicting graft outcome that exceed insignificantly Cys C or sCr areas. CONCLUSIONS: The most reliable predictors of graft outcome were Cys C, sCr and proteinuria. Because Cys C is unavailable in many transplant centers, from the practical point of view, sCr remains the most sensitive predictor of graft outcome.

Correlation of N-terminal pro-B-type natriuretic peptide with clinical parameters in patients with hypertension.

BACKGROUND/AIM: Identification of patients with arterial hypertension and a possible onset of heart failure by determining the concentration of N-terminal pro-B-type natriuretic peptide (NT-proBNP) enables timely intensification of treatment and allows clinicians to prescribe and implement optimal and appropriate care. The aim of this study was to evaluate NT-proBNP in patients with longstanding hypertension and in patients with signs of hypertensive cardiomyopathy. METHODS: The study involved 3 groups, with 50 subjects each: "healthy" persons (control group), patients with hypertension and normal left ventricular systolic function (group 1) and patients with longstanding hypertension and signs of hypertensive cardiomyopathy with impaired left ventricular systolic function (group 2). We measured levels of NT-proBNP, C-reactive protein and creatinine according to the manufacturer's instructions. All the patients were clinically examined including physical examination of the heart with blood pressure, pulse rate, electrocardiogram (ECG) and echocardiogram. RESULTS: Our results showed that the determined parameters generally differed significantly (Student's t-test) among the groups. The mean (+/- SD) values of NT-proBNP in the control group, group 1 and group 2 were: 2.794 (+/- 1.515) pmol/L, 9.575 (+/- 5.449) pmol/L and 204.60 (84,93) pmol/L, respectively. NT-proBNP correlated significantly with the determined parameters both in the group 1 and the group 2. In the group 1, the highest correlation was obtained with C-reactive protein (r = 0.8424). In the group 2, the highest correlation was obtained with ejection fraction (r = -0.9111). NT-proBNP showed progressive increase in proportion to the New York Heart Association (NYHA) classification. The patients in the- group 2 who belonged to the II and III NYHA class had significantly higher levels of NT-proBNP than those in the NYHA class I (ANOVA test, p = 0.001). CONCLUSION: The obtained results suggest that NT-proBNP is a useful biomarker in the treatment of patients with longstanding hypertension who are at risk for heart failure.

Glycogen phosphorylase isoenzyme BB plasma kinetics is not related to myocardial ischemia induced by exercise stress echo test.

BACKGROUND: Glycogen phosphorylase BB (GPBB) is released from cardiac cells during myocyte damage. Previous studies have shown contradictory results regarding the relation of enzyme release and reversible myocardial ischemia. The aim of this study was to determine the plasma kinetics of GPBB as a response to the exercise stress echocardiographic test (ESET), and to define the relationship between myocardial ischemia and enzyme plasma concentrations. METHODS: We studied 46 consecutive patients undergoing ESET, with recent coronary angiography. In all patients, a submaximal stress echo test according to Bruce protocol was performed. Concentration of GPBB was measured in peripheral blood that was sampled 5 min before and 10, 30 and 60 min after ESET. RESULTS: There was significant increase of GPBB concentration after the test (p=0.021). Significant increase was detected 30 min (34.9% increase, p=0.021) and 60 min (34.5% increase, p=0.016) after ESET. There was no significant effect of myocardial ischemia on GPBB concentrations (p=0.126), and no significant interaction between sampling intervals and myocardial ischemia, suggesting a similar release profile of GPBB in ischemic and non-ischemic conditions (p=0.558). Patients in whom ESET was terminated later (stages 4 or 5 of standard Bruce protocol; n=13) had higher GPBB concentrations than patients who terminated ESET earlier (stages 1, 2 or 3; n=33) (p=0.049). Baseline GPBB concentration was not correlated to any of the patients' demographic, clinical and hemodynamic characteristics. CONCLUSIONS: GPBB plasma concentration increases after ESET, and it is not related to inducible myocardial ischemia. However, it seems that GPBB release during ESET might be related to exercise load/duration.

Gammopathy and B lymphocyte clonality in patients with Gaucher type I disease.

INTRODUCTION: We evaluated a novel approach for investigation of lymphocyte dysregulation in Gaucher patients by including determination of IgH and TCR gene rearrangements together with levels of immunoglobulins, natural autoantibodies as well as presence of monoclonal protein. MATERIALS AND METHODS: Measurement of serum immunoglobulins, monoclonal immunoglobulins, selected autoantibodies, as well as analysis of immunoglobulin heavy chain and T cell receptor gene rearrangements. RESULTS: Immunoglobulin disorder was detected in 29.6% patients, 40.7% demonstrated presence of B cell clonality and 44.4% demonstrated presence of autoantibodies. In five patients in our series, the presence of IgH gene rearrangement was the only detectable indicator of B cell dysfunction. TCR gene rearrangements were not found in any of the patients. CONCLUSION: Based on our results, we propose IgH gene rearrangements as a new biomarker for investigation of B cell dysfunction occurring as a complication of Gaucher disease.

[Evaluation of methods for rapid microalbuminuria screening in kidney diseased patients].

INTRODUCTION: One of the criteria for chronic kidney disease detection is determination of microalbuminuria. OBJECTIVE: This analysis was performed to evaluate accuracy of three useful methods for microalbuminuria detection in 24h urine collection and in the morning urine specimen calculated from urine albumin creatinine ratio, or with a dipstick in patients with different kidney diseases or kidney function. METHODS: Microalbuminuria was detected in 74 patients referred to the Outpatient Nephrology Department for kidney function determination or regular nephrology checking. Albumin concentration determined using immunonephelometry was lower than 300 mg/day. Discriminates cutoff values for spot urine test strip and albumin creatinin ratio in predicting 24 h protein'threshold' excretion were determined using ROC analysis. RESULTS: Mean value of 24 h microalbuminuria was 80.3 mg/24 h, and value >30 mg/24 h was present in 71.8% of patient. Correlation coefficients between dipstick microalbuminuria or albumin/creatinine ratio in a spot urine specimen and 24 h microalbuminuria were 0.709 and 0.598 (p<0.0001). For pathological value of 24 h microalbuminuria >30 mg/24 h, the coresponding dipstick microalbuminuria value was > or = 20 mg/L (AUC 0.849, specificity 95%, positive predictive value 97.3%), and > or = 3.55 mg albumin/mmol creatinine ratio (AUC 0.914, specificity 90% and positive predictive value 95.5%). No difference was found between dipstick mikroalbuminuria and albumin/creatinine ratio value. In addition, albumin/creatinine ratio value from 24 h urine was similar to the value obtained from the spot urine sample. CONCLUSION: Obtained results indicated that albuminuria could be determined accurately in spot urine either with the Micral test strip or with albumin creatinine ratio.

Biomarkers in Serbian patients with Gaucher disease.

OBJECTIVES: The aim of the study was to evaluate the efficiency of the biomarkers chitotriosidase (Chito), total acid phosphatase (TACP), angiotensin converting enzyme (ACE) and ferritin in the diagnosis of Gaucher disease (GD) and to assess the utility of biomarkers for monitoring the effects of enzyme replacement therapy (ERT). DESIGN AND METHODS: Forty treatment-naive Gaucher patients were studied. 27/40 GP were put on ERT and monitored every 6 months. RESULTS: The baseline median values of Chito, TACP, ACE and ferritin were highly elevated in GP: 10216 nmol/mL/h, 26.1 U/L, 253 U/L, 515 µg/L, and 555 µg/L, respectively. The only significant difference between mild and moderate GP subgroups is observed for Chito activity (p=0.0116). During ERT, Chito showed the steepest decrease in regard to TACP and ACE, mainly within the first year (71.4%). CONCLUSIONS: Among these biomarkers, Chito proved to be the most useful biomarker for diagnosing GD and monitoring the ERT.

Evaluation of renal damage by urinary beta-trace protein in patients with chronic kidney disease.

BACKGROUND: Beta-trace protein (BTP) was found to be increased in the serum and urine of patients with renal diseases. The aim of this study was to compare the urinary levels of beta-trace protein with levels of other urinary proteins: albumin, beta2-microglobulin (B2M), alpha1-microglobulin (A1M), and cystatin C and to determine its clinical usefulness for detection of renal dysfunction in chronic kidney disease (CKD). METHODS: These markers were measured in 24-hour urine samples from 134 patients with CKD. RESULTS: BTP correlated significantly with A1M (r = 0.871), cystatin C (r = 0.759), total protein (r = 0.684), B2M (r = 0.497), and albumin (r = 0.448) in 24-hour urine samples (P < 0.05). Urinary BTP concentrations in patients with albuminuria below 30 mg/day were significantly lower than in patients with albuminuria above 30 mg/day (P < 0.0001). ROC analysis showed high diagnostic accuracy of BTP for detection of > 30 mg/day albuminuria (AUC 0.908). Urinary BTP was also in significant correlation with the estimated glomerular filtration rate (r = -0.580). CONCLUSIONS: The results of our study suggest that BTP may be a useful and reliable urinary marker of renal dysfunction and may have a place in addition to urinary alpha1-microglobulin and albumin as an alternative marker for tubular damage and the magnitude of renal impairment in patients with chronic kidney disease.

Placental growth factor as short-term predicting biomarker in acute coronary syndrome patients with non-ST elevation myocardial infarction.

OBJECTIVES: The relevance of placental growth factor was analyzed at the admission of patients with acute coronary syndrome (ACS) without ST elevation in prognosis of fatal outcome after 30 days. METHODS: We collected blood samples from 102 ACS patients admitted to the coronary unit with acute chest pain manifesting within the last 12 hours. RESULTS: In all 102 admitted patients, higher values of placental growth factor (PLGF; >13.2 ng/L, average value) indicated a higher risk of fatal outcome (hazard ratio [HR] 2.28, 95% confidence interval [CI] 1.21- 4.76, P = 0.0125). PLGF is an important independent prognostic marker (adjusted HR 2.35, 95% CI 1.98-4.61, P = 0.1338), and this was shown in a multiparameter model, which involved other statistically important markers of relative risk (age >65, gender, and estimated glomerular filtration rate [eGFR]). CONCLUSION: PLGF levels measured at 12 hours of symptom onset and 30 days later may independently predict fatal outcome in patients with ACS without ST elevation.

Utility of placental growth factor for prediction of 30-day adverse event in emergency department population with non-ST elevation acute coronary syndrome.

BACKGROUND: Placental growth factor (PIGF) belongs to the vascular endothelial growth factor family and seems to be an independent biomarker for plaque disruption, ischemia, and thrombosis. Plasma PIGF is rapidly produced in infarcted myocardial tissue during the acute phase of myocardial infarction. In this study, the relevance of PIGF was analyzed at the admission of patients with acute coronary syndrome (ACS) without ST elevation for the prognosis of fatal outcome after 30 days. METHODS: We collected blood samples from 102 ACS patients admitted to the coronary unit with manifesting acute chest pain within the previous 12 hours and measured the levels of PIGF, high-sensitivity C-reactive protein (hsCRP), and cardiac markers: troponin T (cTnT), B-type natriuretic peptide, creatine kinase-MB (CKMB) and CK activity. RESULTS: PIGF, troponin T, and hsCRP levels were significantly higher in non-survivors than in survivors. ROC analysis showed that PIGF had the highest area under ROC curve (AUC, 0.713), but it was not significantly different from AUCs for cTnT and hsCRP. Higher values of PIGF (>13.2 ng/L) pointed towards a higher risk of fatal outcome (HR 2.28; 95 % CI 1.21-4.76; P=0.0125). The multivariable proportional hazards analysis, which had involved other statistically significant markers of relative risk (age and gender), showed that PIGF was an independent prognostic marker (adjusted HR 2.14; 95 % CI 1.08-4.22). CONCLUSIONS: These results confirmed that PIGF is an independent biomarker of short-term adverse outcome in patients with ACS without ST elevation and that plaque instability, represented by PIGF elevation, has an important role in the pathogenesis of future coronary events.

Association of lipid and inflammatory markers with C-reactive protein in cardiovascular risk assessment for primary prevention.

BACKGROUND: High-sensitivity C-reactive protein (hsCRP) has been recognized as an independent marker of cardiovascular risk. Since atherosclerosis is a multifactorial disease, the aim of this study was to determine association between hsCRP and other markers of inflammation and dyslipidemia. MATERIALS AND METHODS: In 242 healthy volunteers, total cholesterol (TC), HDL cholesterol (HDL-C), LDL cholesterol (LDL-C), nonHDL-C, triglycerides (TG) and hsCRP were measured using Olympus AU2700. Apolipoprotein A-I (apoAI), apolipoprotein B (apo B), lipoprotein (a) (Lp(a)), haptoglobin, alpha1-acid glycoprotein (A1AGP), C3 and C4 complement components were determined on Architect c8000, and serum amyloid A (SAA) and fibrinogen on BN II nephelometer and ACL 7000, respectively. RESULTS: Significant (P < 0.05) partial Pearson's correlation coefficients were found between hsCRP and TC (r = 0.172), nonHDL-C (r = 0.182), LDL-C (r = 0.154), apoB (r = 0.167), fibrinogen (r = 0.411), SAA (r = 0.493), A1AGP (r = 0.462), haptoglobin (r = 0.310), C3 (r = 0.349) and C4 (r = 0.371). In multiple regression analysis, BMI, SAA, A1AGP, fibrinogen and nonHDL-C showed independent correlation with hsCRP. Multinomial logistic regression analysis demonstrated that BMI, nonHDL, fibrinogen and SAA were strong predictors of hsCRP concentration. Odds ratios for intermediate and high risk categories compared with the low risk category were 1.177 (1.033-1.341) and 1.289 (1.091-1.523), 1.515 (1.021-2.249) and 2.062 (1.246-3.411), 2.241 (1.268-3.959) and 7.123 (3.259-15.568), and 1.387 (1.179-1.632) and 1.691 (1.397-2.047), for BMI, nonHDL-C, fibrinogen and SAA, respectively. CONCLUSION: The prediction of risk for future cardiac events based on hsCRP concentration, which is the recommended parameter for improving cardiovascular risk stratification, might be complemented with the information about BMI, nonHDL-C, fibrinogen and SAA.

The values of estimated glomerular filtration rate calculated with creatinine and cystatin C based equations in healthy adults.

The glomerular filtration rate (GFR) is widely accepted as the best overall index of kidney function. GFR can be measured as the clearance of exogenous or endogenous filtration markers or clinically estimated from serum concentrations of creatinine or cystatin C. Recently, it has been recommended that an estimated GFR (eGFR) should be reported in addition to the value of filtration markers. In this study, we determined the values of eGFR, based on creatinine and cystatin C equations, in 125 healthy volunteers aged 20-75 years. Creatinine was measured by a kinetic alkaline picrate method on an ARCHITECT ci8200 analyzer (Abbott Diagnostics, Wiesbaden, Germany). Cystatin C was determined by a latex particle-enhanced immunonephelometric assay (BNII, Dade Behring, Marburg, Germany). The eGFR values were calculated for creatinine using the Modification of Diet in Renal Disease (MDRD) study equation and Rule's quadratic equation and for cystatin C according to the equation published by Hoek et al. The reference intervals for eGFRs with MDRD, Rule's quadratic and Hoek's equations were calculated nonparametrically and were determined to be 63.5-124.6 mL/min/1.73 m2, 78.3-139.2 mL/min/1.73 m2 and 72.2-115.6 mL/min/1.73 m2, respectively. According to the US National Kidney Foundation, chronic kidney disease (CKD) can be defined as a GFR < 60 mL/min/1.73 m2. Our results showed that healthy adults had eGFR values > 63.5 mL/min/1.73 m2. However, it is important to note that these normal values overlap with values in stages 1 and 2 of CKD, thus an eGFR greater than 60 mL/min/1.73 m2 does not exclude kidney disease.

Cortisol response to ACTH stimulation correlates with blood interleukin 6 concentration in healthy humans.

OBJECTIVE: Interleukin 6 (IL6) has the ability to influence each level of the hypothalamo-pituitary-adrenocortical (HPA) axis. The aim of the study was to test whether IL6 concentration correlates with the adrenal cortex response to ACTH in healthy humans. We postulated that higher basal IL6 concentration would be associated with the higher cortisol response to the stimulation. DESIGN AND METHODS: Basal IL6 concentration was measured and a low dose (1 microg) ACTH test was performed to assess cortisol response. Twenty-seven apparently healthy subjects (11 male, 16 female, mean age 31.1 years, age range 22-47 years) were included in the study. RESULTS: Data are presented as mean+/-S.E.M. Basal IL6 level was 0.84+/-0.10 pg/ml. Basal cortisol was 351.9+/-18.3 nmol/l. Maximal cortisol during synacthen test was 653.0+/-20.6 nmol/l. Maximal cortisol increment was 301.1+/-20.0 nmol/l. IL6 concentration was not correlated with basal or maximal cortisol concentration, but correlated significantly with cortisol increment (r=0.63, 95% confidence interval) 0.42-0.83). CONCLUSIONS: In our study, we found that higher basal IL6 concentration is associated with the higher cortisol response to ACTH stimulation. Based on previous research and our data, IL6, even in low concentrations and under physiologic conditions, modulates adrenal cortex responsivity to ACTH. Therefore, it seems that immune modulation of HPA axis is also present under physiologic and not only pathologic conditions.

Contribution to the definition of diagnostic criteria for Balkan endemic nephropathy.

BACKGROUND: Diagnostic criteria for Balkan endemic nephropathy (BEN) have not been precisely established. In the present study the predictive value of variables previously proposed as diagnostic criteria for BEN was examined. METHODS: The study involved 182 patients: 98 patients with BEN, 57 patients with other kidney diseases (20 with glomerulonephritis, 17 with tubulointerstitial diseases and 20 with hypertensive nephrosclerosis) and 27 healthy subjects. The BEN group comprised patients who fulfilled criteria for BEN and suspected BEN, together with patients with proteinuria and at least two tubular abnormalities or one tubular abnormality and a history of urothelial tumour. Demographic, clinical, laboratory and ultrasound variables of examined groups were combined in univariate/multivariate logistic regression analysis. RESULTS: Out of 28 analysed variables only urine alpha1-microglobulin (MG) and kidney length were selected as significant predictors in differentiating BEN from other kidney diseases and healthy controls. Using ROC curves the cutoff values of these variables and proteinuria and kidney volume, variables collinear with them, were found. Moderate sensitivity and specificity characterized all these cutoff values except for proteinuria, which provided high sensitivity and specificity in combination of BEN and healthy persons. The predictive value of different combinations of selected variables was not significantly different from the predictive value of each variable individually. CONCLUSIONS: Proteinuria, urine alpha1-MG, kidney length and volume were selected as significant predictors of BEN. Variables related to kidney failure as well as several tubular disorders (urine specific gravity, FENa and TRP) had an insignificant predictive value and could not be used for differential diagnosis of BEN.

Prognostic value of phospholipase A2 group II, C-reactive protein and Simplified Acute Physiological Score II in intensive care patients.

We investigated the prognostic value of group II phospholipase A2 (PLA2-II) and C-reactive protein (CRP) in patients of the intensive care unit (ICU) who developed severe inflammatory reaction. In addition, the biochemical markers were correlated with the Simplified Acute Physiological Score II (SAPS II). Our study comprised 40 patients with multiple injuries and 35 patients with sepsis admitted to the ICU, and assessed during a follow-up of as long as 28 days. There were 18 survivors and 17 non-survivors in the sepsis group, and 22 survivors and 18 nonsurvivors in the group of patients with multiple injuries. In the group of patients with multiple injuries, the intensity of host inflammatory response showed clearly distinct PLA2-II and CRP profiles: the overall levels of PLA2-II and CRP were significantly higher in non-survivors than in survivors. PLA2-II and CRP do not discriminate nonsurvivors from survivors with sepsis. ROC (receiver operating characteristic curve) analysis and the area under the curve (AUC) showed the best classification of polytrauma patients with SAPS II and PLA2-II. In the group of patients with sepsis, only SAPS II appeared to be the most helpful predictive measure regarding patient outcome.

Analytical performance and clinical efficacy for cardiovascular risk estimation of an Olympus immunoturbidimetric high-sensitivity C-reactive protein assay.

Increased C-reactive protein (CRP) concentration within the reference interval (<10.0 mg/L) is a strong predictor of cardiovascular disease (CVD) in apparently healthy adults. Cutoff points for use of CRP in estimating CVD risk are <1, 1-3 and >3 mg/L for low, average and high relative risk, respectively. For measuring CRP concentrations to assess cardiovascular risk, high-sensitivity CRP (hsCRP) assays have been developed. The aim of this study was to evaluate the analytical performance and clinical efficacy for cardiovascular risk estimation of the Olympus immunoturbidimetric latex CRP assay (sensitive application). The comparative method used was the CardioPhase* hsCRP assay, approved by the Food and Drug Administration for use in CVD risk assessment. The imprecision of the Olympus hsCRP assay in the concentration range 0.2-10.0 mg/L was 0.38-8.16% within runs and 3.75-9.63% between runs. For method comparison studies, 194 fresh serum samples were selected to cover the interval 0.15-10.0 mg/L CRP. Comparison of the Dade Behring and Olympus methods was performed using weighted Deming regression analysis (slope 0.99 mg/L, intercept 0.002 mg/L, S(y,x)=0.02 mg/L, r=0.992) and a Bland-Altman relative difference plot (mean difference -0.002%, SD=0.040%). The agreement between the Dade Behring and Olympus methods for relative risk class assignments was 95.4%. Statistical analysis of the agreement between the two methods for each relative risk class showed that the differences between the methods were not statistically significant (p>0.10). Although previous reports found poor performance of the Olympus CRP tests for use in cardiovascular and peripheral vascular risk estimation, our study proved good analytical performance and clinical efficacy of the Olympus hsCRP assay for this use.