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INTRODUCTION Heart Rate Variability (HRV), the variation in time between each heartbeat, is a proxy for vagus nerve function and validated indicator of cardiovascular health. Lower HRV is associated with increased risk of cardiac events and greater vulnerability to psychological stress. Fatigue is a common, frustrating and persistent symptom for IBD patients, with multifaceted mechanisms including nutrient deficiencies, inflammation, and poor sleep. Lower HRV is associated with higher fatigue in patients with cancer and myalgic encephalomyelitis. We postulate lower HRV could be associated with fatigue in IBD patients. METHODS Adults recruited from an outpatient IBD clinic wore a FitBit Inspire 2.0 for 14 days to monitor nighttime total time asleep, restlessness (# of ~30 second awakenings), and HRV (RMSSD in milliseconds (ms)). At baseline, weeks 1 and 2, participants completed the IBD Fatigue Scale and Harvey Bradshaw Index (HBI) or Simple Colitis Clinical Activity Index (SCCAI). Baseline laboratory testing for c-reactive protein (CRP), vitamins B12 and D, and ferritin was done. Averages for HRV and sleep metrics were computed for the 14-day period. Pearson’s correlations assessed relationships between all study variables (any p < .10). Then, partial correlations calculated the relationship between HRV and fatigue scores while controlling for identified confounding variables. We report an interim analysis. RESULTS 41 participants (63% female, 72.5% White, 92.7% non-Hispanic, 61% Crohn’s, 40.02 (SD=14.18) yrs old). Most (>75%) were in remission (HBI/SCCAI< 4). 34% had B12< 400 pg/mL, 20% vitamin D< 30 ng/mL, 54% ferritin< 50 ng/mL. Participants slept an average of 6.6 hours with 24.1(5.48) awakenings per night. All reported fatigue, with 43.9% having severe levels. Fatigue severity and impact remained consistent over the study (p=.106). Average HRV was 31.79(19.0) ms (Range: 11 - 92). Older patients had lower HRV (r= -0.34, p=.025) with no gender differences. Patients who reported more fatigue had higher CRP (r= 0.29), more active IBD symptoms (r= 0.62), and spent more time asleep (r= 0.20). When controlling for these variables and age, IBD patients with lower HRV reported significantly more global fatigue (r= -0.38, p=.041). The relationship between HRV and fatigue was larger for lower HRV and fatigue impact (r= -0.37, p=.047) than fatigue severity (r= -0.30, p=.11). No relationships existed for restlessness, vitamins B12 or D, or ferritin and fatigue. CONCLUSIONS When controlling for other contributors, patients with IBD and lower HRV reported more significant global fatigue and impact of fatigue on daily functioning. Findings may suggest lower HRV increases physical feelings of fatigue and reduces ability to manage fatigue impacts. Increasing HRV, e.g., with biofeedback training, may be a way to improve fatigue symptoms and management in IBD.

The introduction of a second coordination sphere, in the form of a protein scaffold, to synthetic catalysts can be beneficial for their reactivity and substrate selectivity. Here we present semi-native polyacrylamide gel elec-trophoresis (semi-native PAGE) as a rapid screening method for studying metal complex-protein interactions. Such a screening is generally performed using electron spray ionization mass spectrometry (ESI-MS) and/or UV-Vis spectroscopy. Semi-native PAGE analysis has the advantage that it does not rely on spectral changes of the metal complex upon protein interaction and can be applied for high-throughput screening and optimization of complex binding. In semi-native PAGE non-denatured protein samples are loaded on a gel containing sodium dodecyl sulphate (SDS), leading to separation based on differences in structural stability. Semi-native PAGE gel runs of catalyst-protein mixtures were compared to gel runs obtained with native and denaturing PAGE. ESI-MS was additionally realised to confirm protein-complex binding. The general applicability of semi-native PAGE was investigated by screening the binding of various cobalt-and ruthenium-based compounds to three types of haem proteins.

UVR8 is a novel UV-B photoreceptor that regulates a range of plant responses and is already used as a versatile optogenetic tool. Instead of an exogenous chromophore, UVR8 uniquely employs tryptophan side chains to accomplish UV-B photoreception. UV-B absorption by homodimeric UVR8 induces monomerization and hence signaling, but the underlying photodynamic mechanisms are not known. Here, by using a combination of time-resolved fluorescence and absorption spectroscopy from femto- to microseconds, we provide the first experimental evidence for the UVR8 molecular signaling mechanism. The results indicate that tryptophan residues at the dimer interface engage in photoinduced proton coupled electron transfer reactions that induce monomerization.

Light-harvesting antennae of the LHC family form transmembrane three-helix bundles of which two helices are interlocked by conserved arginine-glutamate (Arg-Glu) ion pairs that form ligation sites for chlorophylls. The antenna proteins of photosystem II have an intriguing dual function. In excess light, they can switch their conformation from a light-harvesting into a photoprotective state, in which the excess and harmful excitation energies are safely dissipated as heat. Here we applied magic angle spinning NMR and selective Arg isotope enrichment as a noninvasive method to analyze the Arg structures of the major light-harvesting complex II (LHCII). The conformations of the Arg residues that interlock helix A and B appear to be preserved in the light-harvesting and photoprotective state. Several Arg residues have very downfield-shifted proton NMR responses, indicating that they stabilize the complex by strong hydrogen bonds. For the Arg Cα chemical shifts, differences are observed between LHCII in the active, light-harvesting and in the photoprotective, quenched state. These differences are attributed to a conformational change of the Arg residue in the stromal loop region. We conclude that the interlocked helices of LHCII form a rigid core. Consequently, the LHCII conformational switch does not involve changes in A/B helix tilting but likely involves rearrangements of the loops and helical segments close to the stromal and lumenal ends.

The anisotropic and magic-angle fluorescence decay of the single tryptophan (Trp) residue of melittin, a bee venom peptide, was investigated by time-resolved fluorescence anisotropy using a streak camera setup. The peptide was dissolved either in distilled water or in Hepes/NaOH buffer containing low (10 mM) or high (2 M) concentrations of NaCl, the latter resulting in tetramerized melittin. For melittin in distilled water and low NaCl concentration, two anisotropy decay times were found in the order of similar to50 and similar to800 picoseconds, reflecting, local and overall peptide dynamics, respectively, and for tetramerized melittin, two anisotropy decay times of similar to200 and similar to5500 picoseconds were found. Decay-associated spectra of the isotropic fluorescence decay show three time components in the range of similar to20 picoseconds, similar to500 picoseconds, and similar to3500 picoseconds, respectively. The relative amplitudes of the latter two change upon the self-association of melittin. This change can be explained by the existence of different rotamers of Trp in melittin, of which one is more favored in the melittin tetramer than in the melittin monomer.

We investigated the oligomerization of the core light-harvesting complex (LH1) of Rhodospirillum rubrum from the separated alpha beta BChl(2) subunits (B820) and the oligomerization of the B820 subunit from its monomeric peptides. The full LH1 complex was reversibly associated from B820 subunits by either varying the temperature in the range 277-300 K or by varying the detergent concentration in the buffer from 0.36 to 0.52% n-octyl-beta-D-glucopyranoside. Temperature-induced transition measurements showed hysteresis: raising the temperature induced dissociation of B873 directly into B820 subunits whereas upon recooling an intermediate spectral form was observed with an absorption maximum located around 850 nm. This intermediate form was also observed in detergent-induced transitions. It is speculated that the B850 form is a small aggregate of B820, for instance a dimer. Additionally, during a temperature-mediated transition at low detergent concentration, a set of spectral forms with maxima slightly blue-shifted from 873 nm were observed, possibly due to opened rings with one or only a few alpha beta BChl(2) units missing. The temperature-induced transition of LH1 is discussed in terms of a simple assembly model. It is concluded that a moderately cooperative assembly explains the formation of small aggregates of B820 as well as of incomplete rings. Furthermore, the B820 subunits were reversibly dissociated into the monomeric B777 form by increasing either the temperature or the detergent concentration. Estimations of the enthalpy and entropy changes for the dimeric association reaction of B777 into B820 yielded an enthalpy change of -216 kJ mol(-1) and an entropy change of -0.59 kJ mol(-1)K(-1), at a detergent concentration of 0.8% n-octyl-beta-D-glucopyranoside.

Light-harvesting antennae of the LHC family form transmembrane three-helix bundles of which two helices are interlocked by conserved arginine-glutamate (Arg-Glu) ion pairs that form ligation sites for chlorophylls. The antenna proteins of photosystem II have an intriguing dual function. In excess light, they can switch their conformation from a light-harvesting into a photoprotective state, in which the excess and harmful excitation energies are safely dissipated as heat. Here we applied magic angle spinning NMR and selective Arg isotope enrichment as a noninvasive method to analyze the Arg structures of the major light-harvesting complex II (LHCII). The conformations of the Arg residues that interlock helix A and B appear to be preserved in the light-harvesting and photoprotective state. Several Arg residues have very downfield-shifted proton NMR responses, indicating that they stabilize the complex by strong hydrogen bonds. For the Arg Cα chemical shifts, differences are observed between LHCII in the active, light-harvesting and in the photoprotective, quenched state. These differences are attributed to a conformational change of the Arg residue in the stromal loop region. We conclude that the interlocked helices of LHCII form a rigid core. Consequently, the LHCII conformational switch does not involve changes in A/B helix tilting but likely involves rearrangements of the loops and helical segments close to the stromal and lumenal ends.