Horizontal running inside circular walls of Moon settlements: a comprehensive countermeasure for low-gravity deconditioning?

Long-lasting exposure to low gravity, such as in lunar settlements planned by the ongoing Artemis Program, elicits muscle hypotrophy, bone demineralization, cardio-respiratory and neuro-control deconditioning, against which optimal countermeasures are still to be designed. Rather than training selected muscle groups only, 'whole-body' activities such as locomotion seem better candidates, but at Moon gravity both 'pendular' walking and bouncing gaits like running exhibit abnormal dynamics at faster speeds. We theoretically and experimentally show that much greater self-generated artificial gravities can be experienced on the Moon by running horizontally inside a static 4.7 m radius cylinder (motorcyclists' 'Wall of Death' of amusement parks) at speeds preventing downward skidding. To emulate lunar gravity, 83% of body weight was unloaded by pre-tensed (36 m) bungee jumping bands. Participants unprecedentedly maintained horizontal fast running (5.4-6.5 m s-1) for a few circular laps, with intense metabolism (estimated as 54-74 mlO2 kg-1 min-1) and peak forces during foot contact, inferred by motion analysis, of 2-3 Earth body weight (corresponding to terrestrial running at 3-4 m s-1), high enough to prevent bone calcium resorption. A training regime of a few laps a day promises to be a viable countermeasure for astronauts to quickly combat whole-body deconditioning, for further missions and home return.

NRF2 activation by cysteine as a survival mechanism for triple-negative breast cancer cells.

Triple-negative breast cancer (TNBC) is a very aggressive and heterogeneous group of tumors. In order to develop effective therapeutic strategies, it is therefore essential to identify the subtype-specific molecular mechanisms underlying disease progression and resistance to chemotherapy. TNBC cells are highly dependent on exogenous cystine, provided by overexpression of the cystine/glutamate antiporter SLC7A11/xCT, to fuel glutathione synthesis and promote an oxidative stress response consistent with their high metabolic demands. Here we show that TNBC cells of the mesenchymal stem-like subtype (MSL) utilize forced cystine uptake to induce activation of the transcription factor NRF2 and promote a glutathione-independent mechanism to defend against oxidative stress. Mechanistically, we demonstrate that NRF2 activation is mediated by direct cysteinylation of the inhibitor KEAP1. Furthermore, we show that cystine-mediated NRF2 activation induces the expression of important genes involved in oxidative stress response, but also in epithelial-to-mesenchymal transition and stem-like phenotype. Remarkably, in survival analysis, four upregulated genes (OSGIN1, RGS17, SRXN1, AKR1B10) are negative prognostic markers for TNBC. Finally, expression of exogenous OSGIN1, similarly to expression of exogenous NRF2, can prevent cystine depletion-dependent death of MSL TNBC cells. The results suggest that the cystine/NRF2/OSGIN1 axis is a potential target for effective treatment of MSL TNBCs.

The work to swing limbs in humans versus chimpanzees and its relation to the metabolic cost of walking.

Compared to their closest ape relatives, humans walk bipedally with lower metabolic cost (C) and less mechanical work to move their body center of mass (external mechanical work, WEXT). However, differences in WEXT are not large enough to explain the observed lower C: humans may also do less work to move limbs relative to their body center of mass (internal kinetic mechanical work, WINT,k). From published data, we estimated differences in WINT,k, total mechanical work (WTOT), and efficiency between humans and chimpanzees walking bipedally. Estimated WINT,k is ~ 60% lower in humans due to changes in limb mass distribution, lower stride frequency and duty factor. When summing WINT,k to WEXT, between-species differences in efficiency are smaller than those in C; variations in WTOT correlate with between-species, but not within-species, differences in C. These results partially support the hypothesis that the low cost of human walking is due to the concerted low WINT,k and WEXT.

A novel computational framework for the estimation of internal musculoskeletal loading and muscle adaptation in hypogravity.

Introduction: Spaceflight is associated with substantial and variable musculoskeletal (MSK) adaptations. Characterisation of muscle and joint loading profiles can provide key information to better align exercise prescription to astronaut MSK adaptations upon return-to-Earth. A case-study is presented of single-leg hopping in hypogravity to demonstrate the additional benefit computational MSK modelling has when estimating lower-limb MSK loading. Methods: A single male participant performed single-leg vertical hopping whilst attached to a body weight support system to replicate five gravity conditions (0.17, 0.25, 0.37, 0.50, 1 g). Experimental joint kinematics, joint kinetics and ground reaction forces were tracked in a data-tracking direct collocation simulation framework. Ground reaction forces, sagittal plane hip, knee and ankle net joint moments, quadriceps muscle forces (Rectus Femoris and three Vasti muscles), and hip, knee and ankle joint reaction forces were extracted for analysis. Estimated quadriceps muscle forces were input into a muscle adaptation model to predict a meaningful increase in muscle cross-sectional area, defined in (DeFreitas et al., 2011). Results: Two distinct strategies were observed to cope with the increase in ground reaction forces as gravity increased. Hypogravity was associated with an ankle dominant strategy with increased range of motion and net plantarflexor moment that was not seen at the hip or knee, and the Rectus Femoris being the primary contributor to quadriceps muscle force. At 1 g, all three joints had increased range of motion and net extensor moments relative to 0.50 g, with the Vasti muscles becoming the main muscles contributing to quadriceps muscle force. Additionally, hip joint reaction force did not increase substantially as gravity increased, whereas the other two joints increased monotonically with gravity. The predicted volume of exercise needed to counteract muscle adaptations decreased substantially with gravity. Despite the ankle dominant strategy in hypogravity, the loading on the knee muscles and joint also increased, demonstrating this provided more information about MSK loading. Discussion: This approach, supplemented with muscle-adaptation models, can be used to compare MSK loading between exercises to enhance astronaut exercise prescription.

Metabolic cost and mechanical work of walking in a virtual reality emulator.

PURPOSE: The purpose of this study was to investigate the metabolic cost (C), mechanical work, and kinematics of walking on a multidirectional treadmill designed for locomotion in virtual reality. METHODS: Ten participants (5 females, body mass 67.2 ± 8.1 kg, height 1.71 ± 0.07 m, age 23.6 ± 1.9 years, mean ± SD) walked on a Virtuix Omni multidirectional treadmill at four imposed stride frequencies: 0.70, 0.85, 1.00, and 1.15 Hz. A portable metabolic system measured oxygen uptake, enabling calculation of C and the metabolic equivalent of task (MET). Gait kinematics and external, internal, and total mechanical work (WTOT) were calculated by an optoelectronic system. Efficiency was calculated either as WTOT/C or by summing WTOT to the work against sliding frictions. Results were compared with normal walking, running, and skipping. RESULTS: C was higher for walking on the multidirectional treadmill than for normal walking, running, and skipping, and decreased with speed (best-fit equation: C = 20.2-27.5·speed + 15.8·speed2); the average MET was 4.6 ± 1.4. Mechanical work was higher at lower speeds, but similar to that of normal walking at higher speeds, with lower pendular energy recovery and efficiency; differences in efficiency were explained by the additional work against sliding frictions. At paired speeds, participants showed a more forward-leaned trunk and higher ankle dorsiflexion, stride frequency, and duty factor than normal walking. CONCLUSION: Walking on a multidirectional treadmill requires a higher metabolic cost and different mechanical work and kinematics than normal walking. This raises questions on its use for gait rehabilitation but highlights its potential for high-intensity exercise and physical activity promotion.

Movement in low gravity environments (MoLo) programme-The MoLo-L.O.O.P. study protocol.

BACKGROUND: Exposure to prolonged periods in microgravity is associated with deconditioning of the musculoskeletal system due to chronic changes in mechanical stimulation. Given astronauts will operate on the Lunar surface for extended periods of time, it is critical to quantify both external (e.g., ground reaction forces) and internal (e.g., joint reaction forces) loads of relevant movements performed during Lunar missions. Such knowledge is key to predict musculoskeletal deconditioning and determine appropriate exercise countermeasures associated with extended exposure to hypogravity. OBJECTIVES: The aim of this paper is to define an experimental protocol and methodology suitable to estimate in high-fidelity hypogravity conditions the lower limb internal joint reaction forces. State-of-the-art movement kinetics, kinematics, muscle activation and muscle-tendon unit behaviour during locomotor and plyometric movements will be collected and used as inputs (Objective 1), with musculoskeletal modelling and an optimisation framework used to estimate lower limb internal joint loading (Objective 2). METHODS: Twenty-six healthy participants will be recruited for this cross-sectional study. Participants will walk, skip and run, at speeds ranging between 0.56-3.6 m/s, and perform plyometric movement trials at each gravity level (1, 0.7, 0.5, 0.38, 0.27 and 0.16g) in a randomized order. Through the collection of state-of-the-art kinetics, kinematics, muscle activation and muscle-tendon behaviour, a musculoskeletal modelling framework will be used to estimate lower limb joint reaction forces via tracking simulations. CONCLUSION: The results of this study will provide first estimations of internal musculoskeletal loads associated with human movement performed in a range of hypogravity levels. Thus, our unique data will be a key step towards modelling the musculoskeletal deconditioning associated with long term habitation on the Lunar surface, and thereby aiding the design of Lunar exercise countermeasures and mitigation strategies.

Paneth cells drive intestinal stem cell competition and clonality in aging and calorie restriction.

Calorie restriction has been recently shown to increase intestinal stem cell competition and to reduce mutation fixation in young mice. However, the impact of aging on this process is unknown. By employing Confetti reporter mice, here we show that, unexpectedly, old mice have more intestinal stem cell (ISC) competition than young mice. Moreover, differently from what observed in young mice, calorie restriction, when applied at late-life, decreases this process. Importantly, we also observed a strong correlation between the ISC competition and Paneth cell number. In vivo analysis and in vitro organoid experiments indicated that Paneth cells play a major role in driving intestinal stem cell competition and crypt clonality. Taken together, our results provide evidence that increasing the number of Paneth cells can increase the number of competitive ISCs, representing a valuable therapeutic target to delay fixation of mutated intestinal stem cells.

Rocker-profile design shoes improve pendular energy recovery in walking with no effects on total mechanical work.

Rocker-profile design shoes are commonly used in clinical settings. Such footwear reduces in-shoe pressure over the forefoot area during the gait, and depending on the rocker-profile type (i.e., toe-only, heel negative, or double rocker), affects lower limb kinematics, kinetics, and muscle electromyographic activity. However, whether wearing rocker-profile shoes influence the dynamics of the body centre of mass (BCoM) is unknown. We used a mathematical procedure combining Lissajous contours and Fourier analysis to describe the 3D trajectory of the BCoM in walking with rocker-profile (RollingSole) and flat (Control) shoes at 0.97, 1.25, and 1.53 m s-1 in 30 participants. Harmonics amplitude and phase were compared using linear and circular statistics, respectively. External (Wext), kinematic internal (Wint,k) and total (Wtot) mechanical works, and the mechanical energy fraction recovered from a pendular exchange of potential and kinetic energy were also calculated. RollingSole shoes yielded greater Wext (1-9 %; P < 0.05) and fractional pendular energy exchange (1-8 %; P < 0.01), with lower Wint,k (2-5 %; P < 0.05) and unchanged Wtot (P ≥ 0.30). RollingSole shoes led also to a greater mean height of the BCoM (1-3 %; P < 0.01), and amplitude of the anteroposterior and vertical symmetric, and mediolateral 2nd-to-5th harmonics (1-30 %; P < 0.01). No differences between shoes were found for the harmonics phase (P ≥ 0.14). Our results indicate that RollingSole shoes enhanced an inverted pendulum-like behaviour of the BCoM during walking with no alterations in total mechanical work. This may result from the combination of rocker-profile design and greater BCoM height (through thicker soles) with such shoes, increasing recovery of mechanical energy in step-to-step transitions and mid-stance.

Comparison of three-dimensional body centre of mass trajectories during locomotion through zero- and one-dimensional statistics.

The trajectory of the body centre of mass (BCoM) during locomotion differs through speeds, gaits and pathological states; statistical methods are needed to compare it among different conditions. Here, ten participants walked on treadmill at 1.1 and 1.4 m/s; BCoM trajectories were obtained through stereophotogrammetry and expressed as Fourier series. Trajectories were compared among speeds using (i) zero-dimensional (0D) linear and circular tests for difference on amplitudes and phases of Fourier series harmonic, (ii) one-dimensional statistical parametric mapping (1D-SPM) t-tests on the anteroposterior, mediolateral and vertical axial components of the BCoM trajectory and (iii) 1D-SPM Hotelling's T2 test on the three-dimensional BCoM trajectory. Increasing speed increased the amplitude and decreased the phase of the fundamental (2nd) vertical harmonic. Coherently, the BCoM vertical trajectory featured greater displacement and a forward-shift, combined with greater displacement along the anteroposterior axis. Hotelling's T2 1D-SPM on the whole trajectory featured supra-threshold clusters at the transition between double and single support, and mid of the single support. 0D and 1D test yielded coherent and complementary results: 1D-SPM Hotelling's T2 is suitable to compare whole three-dimensional BCoM trajectories; however, when researchers hypothesize that conditions may impact specific harmonics or axial components, 0D or 1D-SPM t-tests are recommended.

Inertial biometry from commercial 3D body meshes.

Body segments inertial parameters (or, more generally encompassing humans and animal species, inertial biometry), often necessary in kinetics calculations, have been obtained in the past from cadavers, medical 3D imaging, 3D scanning, or geometric approximations. This restricted the inertial archives to a few species. The methodology presented here uses commercial 3D meshes of human and animal bodies, which can be further re-shaped and 'posed', according to an underlying skeletal structure, before processing. The sequence of steps from virtually chopping the mesh to the estimation of inertial parameters of body segments is described. The accuracy of the method is tested by comparing the estimated results to real data published for humans (male and female), horses, and domestic cats. The proposed procedure opens the possibility of remarkably expanding biomechanics research when body size and shape change, or when external tools, such as prosthesis and sport material, take part in biological movement.

Mechanical work as a (key) determinant of energy cost in human locomotion: recent findings and future directions.

NEW FINDINGS: What is the topic of this review? This narrative review explores past and recent findings on the mechanical determinants of energy cost during human locomotion, obtained by using a mechanical approach based on König's theorem (Fenn's approach). What advances does it highlight? Developments in analytical methods and their applications allow a better understanding of the mechanical-bioenergetic interaction. Recent advances include the determination of 'frictional' internal work; the association between tendon work and apparent efficiency; a better understanding of the role of energy recovery and internal work in pathological gait (amputees, stroke and obesity); and a comprehensive analysis of human locomotion in (simulated) low gravity conditions. ABSTRACT: During locomotion, muscles use metabolic energy to produce mechanical work (in a more or less efficient way), and energetics and mechanics can be considered as two sides of the same coin, the latter being investigated to understand the former. A mechanical approach based on König's theorem (Fenn's approach) has proved to be a useful tool to elucidate the determinants of the energy cost of locomotion (e.g., the pendulum-like model of walking and the bouncing model of running) and has resulted in many advances in this field. During the past 60 years, this approach has been refined and applied to explore the determinants of energy cost and efficiency in a variety of conditions (e.g., low gravity, unsteady speed). This narrative review aims to summarize current knowledge of the role that mechanical work has played in our understanding of energy cost to date, and to underline how recent developments in analytical methods and their applications in specific locomotion modalities (on a gradient, at low gravity and in unsteady conditions) and in pathological gaits (asymmetric gait pathologies, obese subjects and in the elderly) could continue to push this understanding further. The recent in vivo quantification of new aspects that should be included in the assessment of mechanical work (e.g., frictional internal work and elastic contribution) deserves future research that would improve our knowledge of the mechanical-bioenergetic interaction during human locomotion, as well as in sport science and space exploration.

Establishment of a fluorescent reporter of RNA-polymerase II activity to identify dormant cells.

Dormancy, a reversible quiescent cellular state characterized by greatly reduced metabolic activity, protects from genetic damage, prolongs survival and is crucial for tissue homeostasis and cellular response to injury or transplantation. Dormant cells have been characterized in many tissues, but their identification, isolation and characterization irrespective of tissue of origin remains elusive. Here, we develop a live cell ratiometric fluorescent Optical Stem Cell Activity Reporter (OSCAR) based on the observation that phosphorylation of RNA Polymerase II (RNApII), a hallmark of active mRNA transcription elongation, is largely absent in dormant stem cells from multiple lineages. Using the small intestinal crypt as a model, OSCAR reveals in real time the dynamics of dormancy induction and cellular differentiation in vitro, and allows the identification and isolation of several populations of transcriptionally diverse OSCARhigh and OSCARlow intestinal epithelial cell states in vivo. In particular, this reporter is able to identify a dormant OSCARhigh cell population in the small intestine. OSCAR therefore provides a tool for a better understanding of dormant stem cell biology.

Superior Properties of N-Acetylcysteine Ethyl Ester over N-Acetyl Cysteine to Prevent Retinal Pigment Epithelial Cells Oxidative Damage.

Oxidative stress plays a key role in the pathophysiology of retinal diseases, including age-related macular degeneration (AMD) and diabetic retinopathy, which are the major causes of irreversible blindness in developed countries. An excess of reactive oxygen species (ROS) can directly cause functional and morphological impairments in retinal pigment epithelium (RPE), endothelial cells, and retinal ganglion cells. Antioxidants may represent a preventive/therapeutic strategy and reduce the risk of progression of AMD. Among antioxidants, N-acetyl-L-cysteine (NAC) is widely studied and has been proposed to have therapeutic benefit in treating AMD by mitigating oxidative damage in RPE. Here, we demonstrate that N-acetyl-L-cysteine ethyl ester (NACET), a lipophilic cell-permeable cysteine derivative, increases the viability in oxidative stressed RPE cells more efficiently than NAC by reacting directly and more rapidly with oxidizing agents, and that NACET, but not NAC, pretreatment predisposes RPE cells to oxidative stress resistance and increases the intracellular reduced glutathione (GSH) pool available to act as natural antioxidant defense. Moreover, we demonstrate the ability of NACET to increase GSH levels in rats' eyes after oral administration. In conclusion, even if experiments in AMD animal models are still needed, our data suggest that NACET may play an important role in preventing and treating retinal diseases associated with oxidative stress, and may represent a valid and more efficient alternative to NAC in therapeutic protocols in which NAC has already shown promising results.

Frictional internal work of damped limbs oscillation in human locomotion.

Joint friction has never previously been considered in the computation of mechanical and metabolic energy balance of human and animal (loco)motion, which heretofore included just muscle work to move the body centre of mass (external work) and body segments with respect to it. This happened mainly because, having been previously measured ex vivo, friction was considered to be almost negligible. Present evidences of in vivo damping of limb oscillations, motion captured and processed by a suited mathematical model, show that: (a) the time course is exponential, suggesting a viscous friction operated by the all biological tissues involved; (b) during the swing phase, upper limbs report a friction close to one-sixth of the lower limbs; (c) when lower limbs are loaded, in an upside-down body posture allowing to investigate the hip joint subjected to compressive forces as during the stance phase, friction is much higher and load dependent; and (d) the friction of the four limbs during locomotion leads to an additional internal work that is a remarkable fraction of the mechanical external work. These unprecedented results redefine the partitioning of the energy balance of locomotion, the internal work components, muscle and transmission efficiency, and potentially readjust the mechanical paradigm of the different gaits.

A slow V̇O2 on-response allows comfortable adoption of aerobically unaffordable walking and running speeds on short stair ascents.

The aim of this study was to investigate the mechanical and metabolic reasons for the spontaneous gait/speed choice when ascending a short flight of stairs, where walking on every step or running on every other step are frequently interchangeable options. The kinematics, oxygen uptake (V̇O2 ), ventilation and heart rate of 24 subjects were sampled during climbing one and two flights of stairs while using the two gaits. Although motor acts were very short in time (5-22 s), metabolic kinetics, extending into the 250 s after the end of climbing, consistently reflected the (metabolic equivalent of the) required mechanical energy and allowed comparison of the two ascent choices: despite a 250% higher mechanical power associated with running, measured [Formula: see text], ventilation and heart rate peaked at only +25% with respect to walking, and in both gaits at much lower values than [Formula: see text] despite predictions based on previous gradient locomotion studies. Mechanical work and metabolic cost of transport, as expected, showed a similar increase (+25%) in running. For stairs up to a height of 4.8 m (30 steps at 53% gradient), running makes us consume slightly more calories than walking, and in both gaits with no discomfort at all. The cardio-respiratory-metabolic responses similarly delay and dampen the replenishment of phosphocreatine stores, which were depleted much faster during the impulsive, highly powered mechanical event, with almost overlapping time courses. This discrepancy between mechanical and metabolic dynamics allows us to afford climbs ranging from almost to very anaerobic, and to interchangeably decide whether to walk or run up a short flight of stairs.

A slow V̇O2 on-response allows to comfortably adopt aerobically unaffordable walking and running speeds in short stairs ascending.

The aim of this paper is to investigate the mechanical and metabolic reasons of the spontaneous gait/speed choice of ascending short flight of stairs, where walking on every step or running on every other step are frequently interchangeable options. Twenty-four subjects' kinematics, oxygen uptake (V̇O2), ventilation and heart rate were sampled during climbing one and two flights of stairs while using the two gaits. Although motor acts were very short in time (5-22 s), metabolic kinetics, extending in the successive 250 s after the end of climbing, consistently reflected the (equivalent of the) needed mechanical energy and allowed to compare the two ascent choices: despite a 250% higher mechanical power associated to running, measured V̇O2, ventilation and heart rate peaked only at +25% with respect to walking, and in both gaits at a much lower values than V̇O2max despite of predictions based on previous gradient locomotion studies. Mechanical work and metabolic cost of transport, as expected, showed similar increase (+25%) in running. For stairs up to 4.8 m tall (30 steps at 53% gradient), running makes us consuming slightly more calories than walking, and in both gaits at no discomfort at all. The cardio-respiratory-metabolic responses similarly delay and damp the replenishing of phosphocreatine stores, which were much faster depleted during the impulsive, highly powered mechanical event, with almost overlapping time courses. Such a discrepancy between mechanical and metabolic dynamics allows to afford almost-to-very anaerobic climbs and to interchangeably decide whether to walk or run up a short flight of stairs.

Comprehensive mechanical power analysis in sprint running acceleration.

Sprint running is a common feature of many sport activities. The ability of an athlete to cover a distance in the shortest time relies on his/her power production. The aim of this study was to provide an exhaustive description of the mechanical determinants of power output in sprint running acceleration and to check whether a predictive equation for internal power designed for steady locomotion is applicable to sprint running acceleration. Eighteen subjects performed two 20 m sprints in a gym. A 35-camera motion capture system recorded the 3D motion of the body segments and the body center of mass (BCoM) trajectory was computed. The mechanical power to accelerate and rise BCoM (external power, Pext ) and to accelerate the segments with respect to BCoM (internal power, Pint ) was calculated. In a 20 m sprint, the power to accelerate the body forward accounts for 50% of total power; Pint accounts for 41% and the power to rise BCoM accounts for 9% of total power. All the components of total mechanical power increase linearly with mean sprint velocity. A published equation for Pint prediction in steady locomotion has been adapted (the compound factor q accounting for the limbs' inertia decreases as a function of the distance within the sprint, differently from steady locomotion) and is still able to predict experimental Pint in a 20 m sprint with a bias of 0.70 ± 0.93 W kg-1 . This equation can be used to include Pint also in other methods that estimate external horizontal power only.