Detection of Airway Colonization by Aspergillus fumigatus by Use of Electronic Nose Technology in Patients with Cystic Fibrosis.

Currently, there is no noninvasive test that can reliably diagnose early invasive pulmonary aspergillosis (IA). An electronic nose (eNose) can discriminate various lung diseases through an analysis of exhaled volatile organic compounds. We recently published a proof-of-principle study showing that patients with prolonged chemotherapy-induced neutropenia and IA have a distinct exhaled breath profile (or breathprint) that can be discriminated with an eNose. An eNose is cheap and noninvasive, and it yields results within minutes. We determined whether Aspergillus fumigatus colonization may also be detected with an eNose in cystic fibrosis (CF) patients. Exhaled breath samples of 27 CF patients were analyzed with a Cyranose 320. Culture of sputum samples defined the A. fumigatus colonization status. eNose data were classified using canonical discriminant analysis after principal component reduction. Our primary outcome was cross-validated accuracy, defined as the percentage of correctly classified subjects using the leave-one-out method. The P value was calculated by the generation of 100,000 random alternative classifications. Nine of the 27 subjects were colonized by A. fumigatus. In total, 3 subjects were misclassified, resulting in a cross-validated accuracy of the Cyranose detecting IA of 89% (P = 0.004; sensitivity, 78%; specificity, 94%). Receiver operating characteristic (ROC) curve analysis showed an area under the curve (AUC) of 0.89. The results indicate that A. fumigatus colonization leads to a distinctive breathprint in CF patients. The present proof-of-concept data merit external validation and monitoring studies.

Small sample sizes in high-throughput miRNA screens: A common pitfall for the identification of miRNA biomarkers.

Since the discovery of microRNAs (miRNAs), circulating miRNAs have been proposed as biomarkers for disease. Consequently, many groups have tried to identify circulating miRNA biomarkers for various types of diseases including cardiovascular disease and cancer. However, the replicability of these experiments has been disappointingly low. In order to identify circulating miRNA candidate biomarkers, in general, first an unbiased high-throughput screen is performed in which a large number of miRNAs is detected and quantified in the circulation. Because these are costly experiments, many of such studies have been performed using a low number of study subjects (small sample size). Due to lack of power in small sample size experiments, true effects are often missed and many of the detected effects are wrong. Therefore, it is important to have a good estimate of the appropriate sample size for a miRNA high-throughput screen. In this review, we discuss the effects of small sample sizes in high-throughput screens for circulating miRNAs. Using data from a miRNA high-throughput experiment on isolated monocytes, we illustrate that the implementation of power calculations in a high-throughput miRNA discovery experiment will avoid unnecessarily large and expensive experiments, while still having enough power to be able to detect clinically important differences.

Individuals with coronary artery disease at a young age and features of the metabolic syndrome have an increased prothrombotic potential.

The relation between coagulation and atherosclerosis has been extensively described, pointing towards a hypercoagulable state in patients with atherosclerosis, especially in young individuals. However, not all studies were conclusive. It is known that the metabolic syndrome (MetS), a risk factor for coronary artery disease (CAD), is related to a higher incidence of thrombo-embolic events. We hypothesised that individuals with CAD at a young age and MetS have an increased prothrombotic potential. It was the study objective to analyse the endogenous thrombin potential (ETP) and related thrombin generation parameters in patients with CAD before the age of 51 in men and 56 in women with and without MetS features and their healthy first-degree relatives. In this case-control study we included 118 CAD patients and 50 first-degree relatives (controls). Parameters of thrombin generation were obtained with calibrated automated thrombinography. An adjusted general linear model (GLM) showed a positive association between the peak thrombin levels and the presence of CAD at a young age. Based on the NCEP criteria we divided our patient group in CAD patients with and without MetS, and compared them to the controls without MetS. We showed that CAD patients with MetS have increased ETP levels, both in comparison with healthy first-degree relatives and with CAD patients without MetS. There were no differences in ETP between patients without MetS and healthy controls. In conclusion, this study shows that individuals with CAD at a young age and MetS features have an increased prothrombotic potential, compared to CAD patients without MetS.