Novel Methodological Tools for Behavioral Interventions: The Case of HRV-Biofeedback. Sham Control and Quantitative Physiology-Based Assessment of Training Quality and Fidelity.

Scientific research on heart rate variability (HRV) biofeedback is burdened by certain methodological issues, such as lack of consistent training quality and fidelity assessment or control conditions that would mimic the intervention. In the present study, a novel sham HRV-biofeedback training was proposed as a credible control condition, indistinguishable from the real training. The Yield Efficiency of Training Index (YETI), a quantitative measure based on the spectral distribution of heart rate during training, was suggested for training quality assessment. A training fidelity criterion derived from a two-step classification process based on the average YETI index and its standard deviation (YETISD) was suggested. We divided 57 young, healthy volunteers into two groups, each subjected to 20 sessions of either real or sham HRV-biofeedback. Five standard HRV measures (standard deviation of the NN (SDNN), root mean square of the standard deviation of the NN (RMSSD), total power, low-frequency (LF), and high-frequency (HF) power) collected at baseline, after 10 and 20 sessions were subjected to analysis of variance. Application of a training fidelity criterion improved sample homogeneity, resulting in a substantial gain in effect sizes of the group and training interactions for all considered HRV indices. Application of methodological amendments, including proper control conditions (such as sham training) and quantitative assessment of training quality and fidelity, substantially improves the analysis of training effects. Although presented on the example of HRV-biofeedback, this approach should similarly benefit other behavioral training procedures that interact with any of the many psychophysiological mechanisms in the human body.

Heart Rate Variability and Accelerometry as Classification Tools for Monitoring Perceived Stress Levels-A Pilot Study on Firefighters.

Chronic stress is the main cause of health problems in high-risk jobs. Wearable sensors can become an ecologically valid method of stress level assessment in real-life applications. We sought to determine a non-invasive technique for objective stress monitoring. Data were collected from firefighters during 24-h shifts using sensor belts equipped with a dry-lead electrocardiograph (ECG) and a three-axial accelerometer. Levels of stress experienced during fire incidents were evaluated via a brief self-assessment questionnaire. Types of physical activity were distinguished basing on accelerometer readings, and heart rate variability (HRV) time series were segmented accordingly into corresponding fragments. Those segments were classified as stress/no-stress conditions. Receiver Operating Characteristic (ROC) analysis showed true positive classification as stress condition for 15% of incidents (while maintaining almost zero False Positive Rate), which parallels the amount of truly stressful incidents reported in the questionnaires. These results show a firm correspondence between the perceived stress level and physiological data. Psychophysiological measurements are reliable indicators of stress even in ecological settings and appear promising for chronic stress monitoring in high-risk jobs, such as firefighting.

FRET-assisted determination of CLN3 membrane topology.

Juvenile neuronal ceroid lipofuscinosis (JNCL) is caused by mutations in the CLN3 gene, which encodes for a putative lysosomal transmembrane protein with thus far undescribed structure and function. Here we investigate the membrane topology of human CLN3 protein with a combination of advanced molecular cloning, spectroscopy, and in silico computation. Using the transposomics cloning method we first created a library of human CLN3 cDNA clones either with a randomly inserted eGFP, a myc-tag, or both. The functionality of the clones was evaluated by assessing their ability to revert a previously reported lysosomal phenotype in immortalized cerebellar granular cells derived from Cln3Δex7/8 mice (CbCln3Δex7/8). The double-tagged clones were expressed in HeLa cells, and FRET was measured between the donor eGFP and an acceptor DyLight547 coupled to a monoclonal α-myc antibody to assess their relative membrane orientation. The data were used together with previously reported experimental data to compile a constrained membrane topology model for hCLN3 using TOPCONS consensus membrane prediction algorithm. Our model with six transmembrane domains and cytosolic N- and C-termini largely agrees with those previously suggested but differs in terms of the transmembrane domain positions as well as in the size of the luminal loops. This finding improves understanding the function of the native hCLN3 protein.