Catalase-peroxidases have a predominant catalatic activity but differ from monofunctional catalases in exhibiting a substantial peroxidatic reaction which has been implicated in the activation of the antitubercular drug isoniazid in Mycobacterium tuberculosis. Hydroperoxidase I of Escherichia coli encoded by katG is a catalase-peroxidase, and residues in its putative active site have been the target of a site directed-mutagenesis study. Variants of residues R102 and H106, on the distal side of the heme, and H267, the proximal side ligand, were constructed, all of which substantially reduced the catalatic activity and, to a lesser extent, the peroxidatic activity. In addition, the heme content of the variants was reduced relative to the wild-type enzyme. The relative ease of heme loss from HPI and a mixture of tetrameric enzymes with 2, 3, and 4 hemes was revealed by mass spectrometry analysis. Conversion of W105 to either an aromatic (F) or aliphatic (I) residue caused a 4-5-fold increase in peroxidatic activity, coupled with a >99% inhibition of catalatic activity. The peroxidatic-to-catalatic ratio of the W105F variant was increased 2800-fold such that compound I could be identified by both electronic and EPR spectroscopy as being similar to the porphyrin cation radical formed in other catalases and peroxidases. Compound I, when generated by a single addition of H(2)O(2), decayed back to the native or resting state within 1 min. When H(2)O(2) was generated enzymatically in situ at low levels, active compound I was evident for up to 2 h. However, such prolonged treatment resulted in conversion of compound I to a reversibly inactivated and, eventually, to an irreversibly inactivated species, both of which were spectrally similar to compound I.

Astronauts returning from space flight and performing Earth-bound activities must rapidly transition from the microgravity-adapted sensorimotor state to that of Earth's gravity. The goal of the current study was to assess locomotor dysfunction and recovery of function after long-duration space flight using a test of functional mobility. Eighteen International Space Station crewmembers experiencing an average flight duration of 185 days performed the functional mobility test (FMT) pre-flight and post-flight. To perform the FMT, subjects walked at a self selected pace through an obstacle course consisting of several pylons and obstacles set up on a base of 10-cm-thick, medium-density foam for a total of six trials per test session. The primary outcome measure was the time to complete the course (TCC, in seconds). To assess the long-term recovery trend of locomotor function after return from space flight, a multilevel exponential recovery model was fitted to the log-transformed TCC data. All crewmembers exhibited altered locomotor function after space flight, with a median 48% increase in the TCC. From the fitted model we calculated that a typical subject would recover to 95% of his/her pre-flight level at approximately 15 days post-flight. In addition, to assess the early motor learning responses after returning from space flight, we modeled performance over the six trials during the first post-flight session by a similar multilevel exponential relation. We found a significant positive correlation between measures of long-term recovery and early motor learning (P < 0.001) obtained from the respective models. We concluded that two types of recovery processes influence an astronaut's ability to re-adapt to Earth's gravity environment. Early motor learning helps astronauts make rapid modifications in their motor control strategies during the first hours after landing. Further, this early motor learning appears to reinforce the adaptive realignment, facilitating re-adaptation to Earth's 1-g environment on return from space flight.

Stochastic resonance (SR) is a phenomenon whereby the response of a non-linear system to a weak periodic input signal is optimized by the presence of a particular non-zero level of noise. Stochastic resonance using imperceptible stochastic vestibular electrical stimulation, when applied to normal young and elderly subjects, has been shown to significantly improve ocular stabilization reflexes in response to whole-body tilt; improved balance performance during postural disturbances and optimize covariance between the weak input periodic signals introduced via venous blood pressure receptors and the heart rate responses. In our study, 15 subjects stood on a compliant surface with their eyes closed. They were given low-amplitude binaural bipolar stochastic electrical stimulation of the vestibular organs in two frequency ranges of 1-2 and 0-30 Hz over the amplitude range of 0 to ±700 μA. Subjects were instructed to maintain an upright stance during 43-s trials, which consisted of baseline (zero amplitude) and stimulation (non-zero amplitude) periods. Measures of stability of the head and trunk using inertial motion unit sensors attached to these segments and the whole body using a force plate were measured and quantified in the mediolateral plane. Using a multivariate optimization criterion, our results show that the low levels of vestibular stimulation given to the vestibular organs improved balance performance in normal healthy subjects in the range of 5-26% consistent with the stochastic resonance phenomenon. In our study, 8 of 15 and 10 of 15 subjects were responsive for the 1-2- and 0-30-Hz stimulus signals, respectively. The improvement in balance performance did not differ significantly between the stimulations in the two frequency ranges. The amplitude of optimal stimulus for improving balance performance was predominantly in the range of ±100 to ±400 μA. A device based on SR stimulation of the vestibular system might be useful as either a training modality to enhance adaptability or skill acquisition, or as a miniature patch-type stimulator that may be worn by people with disabilities due to aging or disease to improve posture and locomotion function.

Low levels of bipolar binaural white noise based imperceptible stochastic electrical stimulation to the vestibular system (stochastic vestibular stimulation, SVS) have been shown to improve stability during balance tasks in normal, healthy subjects by facilitating enhanced information transfer using stochastic resonance (SR) principles. We hypothesize that detection of time-critical sub-threshold sensory signals using low levels of bipolar binaural SVS based on SR principles will help improve stability of walking during support surface perturbations. In the current study 13 healthy subjects were exposed to short continuous support surface perturbations for 60 s while walking on a treadmill and simultaneously viewing perceptually matched linear optic flow. Low levels of bipolar binaural white noise based SVS were applied to the vestibular organs. Multiple trials of the treadmill locomotion test were performed with stimulation current levels varying in the range of 0–1500 μA, randomized across trials. The results show that subjects significantly improved their walking stability during support surface perturbations at stimulation levels with peak amplitude predominantly in the range of 100–500 μA consistent with the SR phenomenon. Additionally, objective perceptual motion thresholds were measured separately as estimates of internal noise while subjects sat on a chair with their eyes closed and received 1 Hz bipolar binaural sinusoidal electrical stimuli. The optimal improvement in walking stability was achieved on average with peak stimulation amplitudes of approximately 35% of perceptual motion threshold. This study shows the effectiveness of using low imperceptible levels of SVS to improve dynamic stability during walking on a laterally oscillating treadmill via the SR phenomenon.