Automation of serum (1→3)-beta-D-glucan testing allows reliable and rapid discrimination of patients with and without candidemia.

Testing for (1→3)-beta-D-glucan (BDG) is used for detection of invasive fungal infection. However, current assays lack automation and the ability to conduct rapid single-sample testing. The Fungitell assay was adopted for automation and evaluated using clinical samples from patients with culture-proven candidemia and from culture-negative controls in duplicate. A comparison with the standard assay protocol was made in order to establish analytical specifications. With the automated protocol, the analytical measuring range was 8-2500 pg/ml of BDG, and precision testing resulted in coefficients of variation that ranged from 3.0% to 5.5%. Samples from 15 patients with culture-proven candidemia and 94 culture-negative samples were evaluated. All culture-proven samples showed BDG values >80 pg/ml (mean 1247 pg/ml; range, 116-2990 pg/ml), which were considered positive. Of the 94 culture-negative samples, 92 had BDG values <60 pg/ml (mean, 28 pg/ml), which were considered to be negative, and 2 samples were false-positive (≥80 pg/ml; up to 124 pg/ml). Results could be obtained within 45 min and showed excellent agreement with results obtained with the standard assay protocol. The automated Fungitell assay proved to be reliable and rapid for diagnosis of candidemia. It was demonstrated to be feasible and cost efficient for both single-sample and large-scale testing of serum BDG. Its 1-h time-to-result will allow better support for clinicians in the management of antifungal therapy.

Mild hypothermia attenuates circulatory and pulmonary dysfunction during experimental endotoxemia.

OBJECTIVE: We tested whether mild hypothermia impacts on circulatory and respiratory dysfunction during experimental endotoxemia. DESIGN: Randomized controlled prospective experimental study. SETTING: Large animal facility, Medical University of Graz, Austria. SUBJECTS: Thirteen anesthetized and mechanically ventilated pigs. INTERVENTIONS: Lipopolysaccharide was administered for 4 hours. With the beginning of lipopolysaccharide infusion, animals were assigned to either normothermia (38°C, n = 7) or mild hypothermia (33°C, n = 6, intravascular cooling) and followed for 8 hours in total. MEASUREMENTS AND MAIN RESULTS: At the end of the protocol, cardiac output was lower in mild hypothermia than in normothermia (4.5 ± 0.4 L/min vs 6.6 ± 0.4 L/min, p < 0.05), but systemic vascular resistance (885 ± 77 dyn·s/cm vs 531 ± 29 dyn·s/cm, p < 0.05) and (Equation is included in full-text article.)(77% ± 6% vs 54% ± 3%, p < 0.05) were higher. Indices of left ventricular contractility in vivo were not different between groups. The high-frequency band in spectral analysis of heart rate variability indicated a better preserved vagal autonomic modulation of sinuatrial node activity in mild hypothermia versus normothermia (87 ± 5 vs 47 ± 5, normalized units, p < 0.05). Plasma norepinephrine levels were elevated compared with baseline in normothermia (2.13 ± 0.27 log pg/mL vs 0.27 ± 0.17 log pg/mL, p < 0.05) but not in mild hypothermia (1.02 ± 0.31 vs 0.55 ± 0.26, p = not significant). At 38°C in vitro, left ventricular muscle strips isolated from the mild hypothermia group had a higher force response to isoproterenol. SaO2 (100% ± 0% vs 92% ± 3%, p < 0.05) and the oxygenation index (PO2/FIO2, 386 ± 52 mm Hg vs 132 ± 32 mm Hg, p < 0.05) were substantially higher in mild hypothermia versus normothermia. Plasma cytokine levels were not consistently different between groups (interleukin 10) or higher (tumor necrosis factor-α and interleukin 6 and 8) during mild hypothermia versus normothermia. CONCLUSION: The induction of mild hypothermia attenuates cardiac and respiratory dysfunction and counteracts sympathetic activation during experimental endotoxemia. This was not associated with lower plasma cytokine levels, indicating a reduction of cytokine responsiveness by mild hypothermia.

The role of CYP3A5 genotypes in dose requirements of tacrolimus and everolimus after heart transplantation.

BACKGROUND: tacrolimus and everolimus are immunosuppressive drugs metabolized by enzymes of the CYP3A subfamily. A common variant of the CYP3A5 gene, CYP3A5*3, results in strongly decreased CYP3A5 activity and has been shown to influence Tacrolimus blood concentrations, but its role for the pharmacogenetics of Everolimus remains unclear. Aim of the study was to examine the role of CYP3A5*3 variant in tacrolimus and everolimus dose and drug levels after heart transplantation. METHODS: The present study comprised 15 patients with Tacrolimus and 30 patients with Everolimus-based maintenance therapy after heart transplantation. CYP3A5 genotypes were determined and correlated with clinical data. RESULTS: In the Tacrolimus group, 13 subjects were CYP3A5 non-expressors (*3/*3 genotype) and two were heterozygous expressors (*1/*3 genotype). Average Tacrolimus dose was significantly higher in subjects expressing CYP3A5 compared to non-expressors. Tacrolimus levels were not significantly different at any point of time. In the Everolimus group, 27 subjects were CYP3A5 non-expressors (*3/*3 genotype) and three were heterozygous expressors (*1/*3). Neither Everolimus dose nor levels were significantly different between CYP3A5 expressors and non-expressors at any point of time. DISCUSSION: We conclude that in adult patients after heart transplantation, CYP3A5 genotypes have a strong influence on Tacrolimus, but not Everolimus dose requirement.

Increased dopamine is associated with the cGMP and homocysteine pathway in female migraineurs.

BACKGROUND: The group of catecholamines, which include dopamine, adrenaline, and noradrenaline, are neurotransmitters which have been considered to play a role in the pathogenesis of migraine. However, the impact of catecholamines, especially dopamine on migraine as well as the exact mechanisms is not clear to date as previous studies have yielded in part conflicting results. OBJECTIVE: This study aimed to produce a comprehensive examination of dopamine in migraineurs. METHODS: Catecholamines and various parameters of the homocysteine, folate, and iron metabolism as well as cyclic guanosine monophosphate (cGMP) and inflammatory markers were determined in 135 subjects. RESULTS: We found increased dopamine levels in the headache free period in female migraineurs but not in male patients. Increased dopamine is associated with a 3.30-fold higher risk for migraine in women. We found no significant effects of aura symptoms or menstrual cycle phases on dopamine levels. Dopamine is strongly correlated with cGMP and the homocysteine-folate pathway. CONCLUSION: We show here that female migraineurs exhibit increased dopamine levels in the headache free period which are associated with a higher risk for migraine.

Human cytomegalovirus load in various body fluids of congenitally infected newborns.

BACKGROUND: Congenital human cytomegalovirus (hCMV) infection is the most common intrauterine viral disease in western countries. Little is known about hCMV virus load in various body fluids of congenitally infected children. OBJECTIVES: To determine virus load in various body fluids. To assess the impact of hCMV virus load to predict the outcome of congenitally infected newborns and efficacy of antiviral therapy. STUDY DESIGN: Cord vein blood, urine, and cerebrospinal fluid (CSF) of congenitally hCMV-infected children were investigated and hCMV load was determined by quantitative polymerase chain reaction (PCR). Fourteen of 30 children had clinical symptoms and/or pathological laboratory results and 16 had none of them at birth. Ganciclovir was given to 21 children (10 of them with symptoms, 11 of them without symptoms). Viral load before and after therapy was measured. RESULTS: There was a significant difference between median virus load in cord vein blood (2.3 x 10(3) copies per ml) and in urine (4.2 x 10(5) copies per ml; P<0.001) at diagnosis of congenital hCMV infection. At that time, no significant difference of virus load was found between the various groups (symptomatic vs. asymptomatic; with therapy vs. without therapy), neither in serum nor in urine. Comparing median virus load in urine before (3.0 x 10(5) copies per ml) and after therapy (2.0 x 10(3) copies per ml), a significant decrease was observed (P<0.001). Virus load in CSF was always found to be less than 400 copies per ml, and only those children with symptoms showed a positive result. CONCLUSION: At birth, virus load in urine seems to be superior to that in cord vein blood to reflect the situation in the organs precisely. As predicting factor for the risk of developing symptoms, only hCMV detection in the CSF appears to be promising. The significant decrease of virus load in children with therapy may reflect the efficacy of therapy. Studies including a greater number of children are needed.

Left ventricular diastolic dysfunction during acute myocardial infarction: effect of mild hypothermia.

BACKGROUND: Mild hypothermia (MH) decreases infarct size and mortality in experimental reperfused myocardial infarction, but may potentiate ischaemia-induced left ventricular (LV) diastolic dysfunction. METHODS: In anaesthetized pigs (70 ± 2 kg), polystyrol microspheres (45 µm) were infused repeatedly into the left circumflex artery until cardiac power output decreased >40%. Then, pigs were assigned to normothermia (NT, 38.0°C, n=8) or MH (33.0°C, n=8, intravascular cooling) and followed for 6h (CME 6h). p<0.05 vs baseline, †p<0.05 vs NT. RESULTS: In NT, cardiac output (CO) decreased from 6.2 ± 0.3 to 3.4 ± 0.2 l/min, and heart rate increased from 89 ± 4 to 101 ± 6 bpm. LV end-diastolic volume fell from 139 ± 8 to 64 ± 4 ml, while LV ejection fraction remained constant (49 ± 1 vs 53 ± 4%). The corresponding end-diastolic pressure-volume relationship was progressively shifted leftwards, reflecting severe LV diastolic dysfunction. In MH, CO fell to a similar degree. Spontaneous bradycardia compensated for slowed LV relaxation, and the leftward shift of the end-diastolic pressure-volume relationship was less pronounced during MH. MH increased systemic vascular resistance, such that mean aortic pressure remained higher in MH vs NT (69 ± 2† vs 54 ± 4 mm Hg). Mixed venous oxygen saturation at CME 6h was higher in MH than in NT (59 ± 4† vs 42 ± 2%) due to lowered systemic oxygen demand during cooling. CONCLUSION: We conclude that (i) an acute loss of end-diastolic LV compliance is a major component of acute cardiac pump failure during experimental myocardial infarction, and that (ii) MH does not potentiate this diastolic LV failure, but stabilizes haemodynamics and improves systemic oxygen supply/demand imbalance by reducing demand.

Trunk weighted obesity, cholesterol levels and low grade inflammation are main determinants for enhanced thrombin generation.

OBJECTIVE: Endogenous thrombin generation (ETP) may be critically involved in obesity associated thromboembolism. METHODS: Three hundred and one participants of the STyrian Juvenile OBesity (STYJOBS)/Early DEteCTion of Atherosclerosis (EDECTA) study cohort (age, 16-58years) were analysed. ETP was measured by the new CE-IVD marked Siemens-Innovance(®) ETP test on a BCS-XP analyser, and correlated to clinical findings and extended lipometry-based anthropometric data, biomarkers, and coagulation parameters. RESULTS: In the overweight/obese study group, ETP and fibrinogen levels were significantly higher compared to controls (p<0.001). In a multiple stepwise regression including all subjects, subcutaneous adipose tissue thickness of upper back, cholesterol and ultrasensitive C-reactive protein were the best predictors for ETP. CONCLUSION: Trunk weighted obesity together with low grade inflammation and hypercholesterolemia enhance thrombin generation.

Link between leptin and interleukin-6 levels in the initial phase of obesity related inflammation.

The mechanisms underlying the pathogenesis of obesity-related atherosclerosis remain to be clarified. To investigate the preclinical phase, interleukin-6 (IL-6) plasma levels were analyzed together with clinical, anthropometric, inflammatory, and metabolic variables in a well-defined cohort of 677 young and middle-aged overweight/obese and normal-weight subjects. In the juvenile and adult overweight/obese study group, IL-6 levels were increased significantly compared with normal-weight, age-matched controls (P < 0.001). In both juveniles and adults, higher levels of IL-6 were observed in obese compared with overweight participants. Subjects with metabolic syndrome (MS) had significantly higher IL-6 levels than those without MS. In juveniles, leptin, and in adults, the waist-to-height ratio, turned out to be the best predictor of IL-6 plasma levels in a multiple stepwise regression model. Taken together, in every age group, interleukin-6 is associated positively with the grade of overweight. Interestingly, leptin, which is the best known adipokine, is associated predictively with interleukin-6 plasma levels only in juveniles, which may indicate an important role of this molecule in the initiation of obesity-related inflammation.

High density lipoprotein cholesterol level is a robust predictor of lipid peroxidation irrespective of gender, age, obesity, and inflammatory or metabolic biomarkers.

BACKGROUND: Obesity related dyslipidemia, chronic inflammation and oxidative stress were associated with atherosclerotic sequels. We analysed oxidized low-density lipoprotein (oxLDL) plasma levels of 797 participants of the STyrian Juvenile OBesity (STYJOBS) / Early DEteCTion of Atherosclerosis (EDECTA) Study cohort aged from 5 to 50 years. The rationale of STYJOBS/EDECTA is to investigate the preclinical phase of obesity by a well defined cohort of young and middle aged overweight/obese and normal weight subjects. METHODS AND RESULTS: Plasma oxLDL was analysed by ELISA (Mercodia, Sweden). In the overweight/obese (OW/OB) study group, oxLDL levels were significantly increased compared to normal weighted controls (p<0.001). Probands with metabolic syndrome (MS) had significantly higher oxLDL levels than probands without MS; between overweight and obese participants, and between females and males, no significant difference was seen. In a multiple stepwise regression analysis including all study subjects, age, gender, anthropometric data, presence of metabolic syndrome, systolic, diastolic blood pressure, carotis communis intima media thickness, lipids, adipokines, metabolic, and inflammatory biomarkers, decreased high-density lipoprotein (HDL-cholesterol) and increased total cholesterol were the best predictors for increased oxLDL levels. CONCLUSION: Decreased HDL-cholesterol is an important determinant of lipid peroxidation irrespective of obesity, age, gender, SAT distribution, and inflammatory/metabolic biomarkers.