Respiratory Variability, Sighing, Anxiety, and Breathing Symptoms in Low- and High-Anxious Music Students Before and After Performing.

Music performance anxiety (MPA) is a major problem for music students. It is largely unknown whether music students who experience high or low anxiety differ in their respiratory responses to performance situations and whether these co-vary with self-reported anxiety, tension, and breathing symptoms. Affective processes influence dynamic respiratory regulation in ways that are reflected in measures of respiratory variability and sighing. This study had two goals. First, we determined how measures of respiratory variability, sighing, self-reported anxiety, tension, and breathing symptoms vary as a function of the performance situation (practice vs. public performance), performance phase (pre-performance vs. post-performance), and the general MPA level of music students. Second, we analyzed to what extent self-reported anxiety, tension, and breathing symptoms co-vary with the respiratory responses. The participants were 65 university music students. We assessed their anxiety, tension, and breathing symptoms with Likert scales and recorded their respiration with the LifeShirt system during a practice performance and a public performance. For the 10-min periods before and after each performance, we computed number of sighs, coefficients of variation (CVs, a measure of total variability), autocorrelations at one breath lag (ARs(1), a measure of non-random variability) and means of minute ventilation (V'E), tidal volume (VT), inspiration time (TI), and expiration time (TE). CVs and sighing were greater whereas AR(1) of V'E was lower in the public session than in the practice session. The effect of the performance situation on CVs and sighing was larger for high-MPA than for low-MPA participants. Higher MPA levels were associated with lower CVs. At the within-individual level, anxiety, tension, and breathing symptoms were associated with deeper and slower breathing, greater CVs, lower AR(1) of V'E, and more sighing. We conclude that respiratory variability and sighing are sensitive to the performance situation and to musicians' general MPA level. Moreover, anxiety, tension, breathing symptoms, and respiratory responses co-vary significantly in the context of music performance situations. Respiratory monitoring can add an important dimension to the understanding of music performance situations and MPA and to the diagnostic and intervention outcome assessments of MPA.

Robust synchronization of coupled circadian and cell cycle oscillators in single mammalian cells.

Circadian cycles and cell cycles are two fundamental periodic processes with a period in the range of 1 day. Consequently, coupling between such cycles can lead to synchronization. Here, we estimated the mutual interactions between the two oscillators by time-lapse imaging of single mammalian NIH3T3 fibroblasts during several days. The analysis of thousands of circadian cycles in dividing cells clearly indicated that both oscillators tick in a 1:1 mode-locked state, with cell divisions occurring tightly 5 h before the peak in circadian Rev-Erbα-YFP reporter expression. In principle, such synchrony may be caused by either unidirectional or bidirectional coupling. While gating of cell division by the circadian cycle has been most studied, our data combined with stochastic modeling unambiguously show that the reverse coupling is predominant in NIH3T3 cells. Moreover, temperature, genetic, and pharmacological perturbations showed that the two interacting cellular oscillators adopt a synchronized state that is highly robust over a wide range of parameters. These findings have implications for circadian function in proliferative tissues, including epidermis, immune cells, and cancer.

Stimulus-induced modulation of transcriptional bursting in a single mammalian gene.

Mammalian genes are often transcribed discontinuously as short bursts of RNA synthesis followed by longer silent periods. However, how these "on" and "off" transitions, together with the burst sizes, are modulated in single cells to increase gene expression upon stimulation is poorly characterized. By combining single-cell time-lapse luminescence imaging with stochastic modeling of the time traces, we quantified the transcriptional responses of the endogenous connective tissue growth factor gene to different physiological stimuli: serum and TGF-ß1. Both stimuli caused a rapid and acute increase in burst sizes. Whereas TGF-ß1 showed prolonged transcriptional activation mediated by an increase of transcription rate, serum stimulation resulted in a large and temporally tight first transcriptional burst, followed by a refractory period in the range of hours. Our study thus reveals how different physiological stimuli can trigger kinetically distinct transcriptional responses of the same gene.

Structural and functional insights into IκB-α/HIV-1 Tat interaction.

Protein-protein interactions play fundamental roles in physiological and pathological biological processes. The characterization of the structural determinants of protein-protein recognition represents an important step for the development of molecular entities able to modulate these interactions. We have recently found that IκB-α (nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha) blocks the HIV-1 expression and replication in a NF-κB-independent manner by directly binding to the virus-encoded Tat transactivator. Here, we report the evaluation of the entity of binding of IκB-α to Tat through in vitro Surface Plasmon Resonance assay. Moreover, by designing and characterizing a set of peptides of the C-terminus region of IκB-α, we show that the peptide corresponding to the IκB-α sequence 262-287 was able to bind to Tat with high affinity (300 nM). The characterization of a number of IκB-α-based peptides also provided insights into their intrinsic folding properties. These findings have been corroborated by mutagenesis studies on the full-length IκB-α, which unveil that different IκB-α residues are involved in NF-κB or Tat recognition.