The Accumulation of Phenyllactic Acid Impairs Host Glutamine Metabolism and Inhibits African Swine Fever Virus Replication: A Novel Target for the Development of Anti-ASFV Drugs.

African swine fever (ASF) is a highly contagious and hemorrhagic disease caused by infection with the African swine fever virus (ASFV), resulting in a mortality rate of up to 100%. Currently, there are no effective treatments and commercially available vaccines for ASF. Therefore, it is crucial to identify biochemicals derived from host cells that can impede ASFV replication, with the aim of preventing and controlling ASF. The ASFV is an acellular organism that promotes self-replication by hijacking the metabolic machinery and biochemical resources of host cells. ASFV specifically alters the utilization of glucose and glutamine, which are the primary metabolic sources in mammalian cells. This study aimed to investigate the impact of glucose and glutamine metabolic dynamics on the rate of ASFV replication. Our findings demonstrate that ASFV infection favors using glutamine as a metabolic fuel to facilitate self-replication. ASFV replication can be substantially inhibited by blocking glutamine metabolism. The metabolomics analysis of the host cell after late-stage ASFV infection revealed a significant disruption of normal glutamine metabolic pathways due to the abundant expression of PLA (phenyllactic acid). Pretreatment with PLA also inhibited ASFV proliferation and glutamine consumption following infection. The metabolomic analysis also showed that PLA pretreatment greatly slowed down the metabolism of amino acids and nucleotides that depend on glutamine. The depletion of these building blocks directly hindered the replication of ASFV by decreasing the biosynthetic precursors produced during the replication of ASFV's progeny virus. These findings provide valuable insight into the possibility of pursuing the development of antiviral drugs against ASFV that selectively target metabolic pathways.

The Effects of Selected Extraction Methods and Natural Deep Eutectic Solvents on the Recovery of Active Principles from Aralia elata var. mandshurica (Rupr. & Maxim.) J. Wen: A Non-Targeted Metabolomics Approach.

The methods and solvents employed in routine extraction protocols essentially impact the composition of the resulting extracts, i.e., the relative abundances of individual biologically active metabolites and the quality and stability of the isolates. Natural deep eutectic solvents (NADESs) represent a new class of environmentally friendly solvents, which are recognized as promising extractants alternative to conventional organic liquids. However, their relative efficiencies when applied in different extraction workflows are still poorly characterized. Therefore, here, we compare the potential of three extraction methods for the extraction of biologically active natural products from Aralia elata var. mandshurica with selected natural deep eutectic solvents (NADESs) using a non-targeted metabolomics approach. The non-targeted metabolite profiling relied on reversed-phase ultra-high-performance liquid chromatography-high-resolution mass spectrometry (RP-UHPLC-HR-MS). The roots of A. elata were extracted by maceration, ultrasound-assisted extraction (UAE), and vibrocavitation-assisted extraction (VAE). Principal component analysis (PCA) revealed a clear separation of the extracts obtained with the three extraction methods employed with NADES1 (choline chloride/malic acid) and NADES2 (sorbitol/malic acid/water). Based on the results of the hierarchical clustering analysis obtained for the normalized relative abundances of individual metabolites and further statistical evaluation with the t-test, it could be concluded that NADES1 showed superior extraction efficiency for all the protocols addressed. Therefore, this NADES was selected to compare the efficiencies of the three extraction methods in more detail. PCA followed by the t-test yielded only 3 metabolites that were more efficiently extracted by maceration, whereas 46 compounds were more abundant in the extracts obtained by VAE. When VAE and UAE were compared, 108 metabolites appeared to be more abundant in the extracts obtained by VAE, whereas only 1 metabolite was more efficiently recovered by UAE. These facts clearly indicate the advantage of the VAE method over maceration and UAE. Seven of the twenty-seven metabolites tentatively identified by tandem mass spectrometry (MS/MS) were found in the roots of A. elata for the first time. Additional studies are necessary to understand the applicability of VAE for the extraction of other plant materials.

Astrocytes: Lessons Learned from the Cuprizone Model.

A diverse array of neurological and psychiatric disorders, including multiple sclerosis, Alzheimer's disease, and schizophrenia, exhibit distinct myelin abnormalities at both the molecular and histological levels. These aberrations are closely linked to dysfunction of oligodendrocytes and alterations in myelin structure, which may be pivotal factors contributing to the disconnection of brain regions and the resulting characteristic clinical impairments observed in these conditions. Astrocytes, which significantly outnumber neurons in the central nervous system by a five-to-one ratio, play indispensable roles in the development, maintenance, and overall well-being of neurons and oligodendrocytes. Consequently, they emerge as potential key players in the onset and progression of a myriad of neurological and psychiatric disorders. Furthermore, targeting astrocytes represents a promising avenue for therapeutic intervention in such disorders. To gain deeper insights into the functions of astrocytes in the context of myelin-related disorders, it is imperative to employ appropriate in vivo models that faithfully recapitulate specific aspects of complex human diseases in a reliable and reproducible manner. One such model is the cuprizone model, wherein metabolic dysfunction in oligodendrocytes initiates an early response involving microglia and astrocyte activation, culminating in multifocal demyelination. Remarkably, following the cessation of cuprizone intoxication, a spontaneous process of endogenous remyelination occurs. In this review article, we provide a historical overview of studies investigating the responses and putative functions of astrocytes in the cuprizone model. Following that, we list previously published works that illuminate various aspects of the biology and function of astrocytes in this multiple sclerosis model. Some of the studies are discussed in more detail in the context of astrocyte biology and pathology. Our objective is twofold: to provide an invaluable overview of this burgeoning field, and, more importantly, to inspire fellow researchers to embark on experimental investigations to elucidate the multifaceted functions of this pivotal glial cell subpopulation.

Metabolic cost of walking with electromechanical ankle exoskeletons under proportional myoelectric control on a treadmill and outdoors.

Lower limb robotic exoskeletons are often studied in the context of steady state treadmill walking in a laboratory environment. However, the end goal for exoskeletons is to be used in real world, complex environments. To reach the point that exoskeletons are openly adopted into our everyday lives, we need to understand how the human and robot interact outside of a laboratory. Metabolic cost is often viewed as a gold standard metric for measuring exoskeleton performance but is rarely used to evaluate performance at non steady state walking outside of a laboratory. In this study, we tested the effects of robotic ankle exoskeletons under proportional myoelectric control on the cost of transport of walking both inside on a treadmill and outside overground. We hypothesized that walking with the exoskeletons would lead to a lower cost of transport compared to walking without them both on a treadmill and outside. We saw no significant increases or decreases in cost of transport or exoskeleton mechanics when walking with the exoskeletons compared to walking without them both on a treadmill and outside. We saw a strong negative correlation between walking speed and cost of transport when walking with and without the exoskeletons. In the future, research should consider how performing more difficult tasks, such as incline and loaded walking, affects the cost of transport while walking with and without robotic ankle exoskeletons. The value of this study to the literature is that it emphasizes the importance of both hardware dynamics and controller design towards reducing metabolic cost of transport with robotic ankle exoskeletons. When comparing our results to other studies using the same hardware with different controllers or very similar controllers with different hardware, there are a wide range of outcomes as to metabolic benefit.

A metabolomic and proteomic study to elucidate the molecular mechanisms of immunotherapy resistance in patients with oesophageal squamous cell carcinoma.

Systemic chemotherapy, the standard first-line treatment option for patients with advanced oesophageal squamous cell carcinoma (OSCC), results in a median survival of ~1 year. Immune checkpoint inhibitors are a breakthrough oncology treatment option; however, most patients with advanced OSCC develop primary and acquired resistance to programmed death receptor-1 (PD-1) monoclonal antibody, severely affecting their prognosis. Therefore, there is an urgent need to investigate the molecular mechanism underlying resistance to treatment. The present study aimed to explore the mechanism of resistance to PD-1 monoclonal antibody. Plasma samples were collected from patients with OSCC treated with immunotherapy, who achieved pathological response/partial response (CR/PR) or stable disease/progressive disease (SD/PD) after the fourth treatment cycle. TM-widely targeted metabolomics, widely targeted lipidomics, and DIA proteomics assays were performed. Differential metabolites were screened based on fold change (FC) ≥1.5 or ≤0.67 and a VIP ≥1; differential proteins were screened based on FC >1.5 or <0.67 and P<0.05. The identified metabolites were annotated and mapped using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway databases. The differential proteins were annotated to the Gene Ontology and KEGG pathway databases. A correlation network diagram was drawn using differential expressed proteins and metabolites with (Pearson correlation coefficient) r>0.80 and P<0.05. Finally, 197 and 113 differential metabolites and proteins were screened, respectively, in patients with CR/PR and SD/PD groups. The KEGG enrichment analysis revealed that all of these metabolites and proteins were enriched in cholesterol metabolism and in the NF-κB and phospholipase D signalling pathways. The present study is the first to demonstrate that PD-1 inhibitor resistance may be attributed to cholesterol metabolism or NF-κB and phospholipase D signalling pathway activation. This finding suggests that targeting these signalling pathways may be a promising novel therapeutic approach in OSCC which may improve prognosis in patients undergoing immunotherapy.

Nontargeted metabolomics integrated with 1 H NMR and LC-Q-TOF-MS/MS methods to depict a more comprehensive metabolic profile in response to chrysosplenetin and artemisinin co-treatment against artemisinin-sensitive and -resistant Plasmodium berghei K173.

Our previous work revealed mutual and specific metabolites/pathways in artemisinin-sensitive and -resistant Plasmodium berghei K173-infected mice. In this study, we further investigated whether chrysosplenetin, a candidate chemical to prevent artemisinin resistance, can regulate these metabolites/pathways by integrating nontargeted metabolomics with 1 H NMR and LC-Q-TOF-MS/MS spectrum. The nuclear magnetic resonance method generated specifically altered metabolites in response to co-treatment with chrysosplenetin, including: the products of glycolysis such as glucose, pyruvate, lactate and alanine; taurine, closely associated with liver injury; arginine and proline as essential amino acids for parasites; TMAO, a biomarker for dysbacteriosis and renal function; and tyrosine, which is used to generate levodopa and dopamine and may improve the torpor state of mice. Importantly, we noticed that chrysosplenetin might depress the activated glycolysis induced by sensitive parasites, but oppositely promoted the inhibited glycolysis to generate more lactate, which suppresses the proliferation of resistant parasites. Moreover, chrysosplentin possibly disturbs the heme biosynthetic pathway in mitochondria. The MS method yielded changed coenzyme A, phosphatidylcholine and ceramides, closely related to mitochondria ß-oxidation, cell proliferation, differentiation and apoptosis. These two means shared no overlapped metabolites and formed a more broader metabolic map to study the potential mechanisms of chrysosplenetin as a promising artemisinin resistance inhibitor.

Differences in Muscle Demand and Joint Contact Forces Between Running and Skipping.

Skipping has been proposed as a viable cross-training exercise to running due to its lower knee contact forces and higher whole-body energy expenditure. However, how individual muscle forces, energy expenditure, and joint loading are affected by differences in running and skipping mechanics remains unclear. The purpose of this study was to compare individual muscle forces, energy expenditure, and lower extremity joint contact forces between running and skipping using musculoskeletal modeling and simulations of young adults (n = 5) performing running and skipping at 2.5 m·s-1 on an instrumented treadmill. In agreement with previous work, running had greater knee and patella contact forces than skipping which was accompanied by greater knee extensor energetic demand. Conversely, skipping had greater ankle contact forces and required greater energetic demand from the uniarticular ankle plantarflexors. There were no differences in hip contact forces between gaits. These findings further support skipping as a viable alternative to running if the primary goal is to reduce joint loading at the commonly injured patellofemoral joint. However, for those with ankle injuries, skipping may not be a viable alternative due to the increased ankle loads. These findings may help clinicians prescribe activities most appropriate for a patient's individual training or rehabilitation goals.

Biomarkers and Cellular Biology in Perioperative Medicine.

Surgical procedures alter tissue integrity; are associated with pain and activation of the sympathetic nervous system; and sometimes, cause exposure to foreign materials used during the surgery or implanted perioperatively [...].

Bone healing: Advances in biology and technology.

Fracture healing is a complex cascade of cellular and molecular processes. These processes require the appropriate cellular and molecular environment to ensure the restoration of skeletal stability and resolution of inflammation. In order for fracture healing to occur, the necessary building blocks for bone metabolism and synthesis must be supplied through proper nutrition. Pharmacologic therapies aimed at modulating the inflammatory response to fractures have the potential to interfere with the synthesis of molecules needed for the production of bone. Infection can interfere with, and even prevent normal fracture healing from occurring. Cellular and genetic treatment strategies are actively being developed to target deficiencies, and bridge gaps that can influence how fractures heal. Evolving technologies, including nutritional supplementation, pharmacotherapies, antibiotics, surgical techniques, as well as genetic and cellular therapies, have the potential to enhance, optimize, and even revolutionize the process of fracture healing.

Metabolomic-proteomic combination analysis reveals the targets and molecular pathways associated with hydrogen sulfide alleviating NAFLD.

AIMS: Nonalcoholic fatty liver disease (NAFLD) is the most common form of chronic liver disease worldwide. Exogenous H2S has been shown to effectively mitigate NAFLD, although little is known about the underlying targets and molecular mechanisms. METHODS: C57BL/6 mice were fed with normal fat diet (NFD) or high fat diet (HFD) for a total 16 weeks, and HFD-fed mice were treated with saline or NaHS beginning in 12th week. The combination analysis of metabolomics and proteomics of liver tissues was firstly performed to discover the candidate targets and potential molecular pathways involved in H2S mitigating the NAFLD. KEY FINDINGS: Compared with NaCl, H2S relieved NAFLD by reducing liver weight, body weight and lipid accumulation in liver, and improving liver pathology and serum biochemical parameters. There were 40 overlapping metabolites in the intersection analysis between comparative analysis of HFD + NaCl vs NFD and HFD + NaHS vs HFD + NaCl based on liver metabolomics. Moreover, a total of 58 proteins were obtained whose changes were reversed after treatment with H2S. A combined analysis of liver metabolomics and proteomics was then conducted, revealing 8 shared molecular pathways, as well as the enrichment of unsaturated fatty acids. In addition, Plin2 may also be a potential target of H2S via the regulation of lipid droplet degradation in alleviating NAFLD. SIGNIFICANCE: We performed the first study combining metabolomics and proteomics to explore the mechanisms behind the alleviation of NAFLD by H2S. Our results not only provide evidence that H2S alleviates NAFLD but also reveals its possible molecular mechanisms and targets.

Brief communication: Maximum ingested bite size in captive western lowland gorillas (Gorilla gorilla gorilla).

OBJECTIVES: Previously, we found that maximum ingested bite size (Vb ), the largest piece of food an animal can consume without biting it into smaller pieces first, isometrically scales relative to body size in strepsirrhines and with negative allometry in anthropoids. In the current study, we rectify the omission of great apes from the earlier sample to now characterize the Vb of the entire size-range of the order. MATERIALS AND METHODS: Five gorillas (Gorilla gorilla gorilla-G. g. gorilla) were studied to ascertain Vb in relation to the mechanical properties of five foods. RESULTS: Gorilla Vb ranged from 166.38 cm3 (for the least obdurate food: watermelon) to 8 cm3 (for the most obdurate food: turnip), with an average Vb of 33.50 cm3 across all food types. CONCLUSIONS: When these data were compared to those from our previous studies, we found that gorillas consumed relatively slightly smaller volumes of food compared to the trend found across primates. However, because the more frugivorous gorillas consumed relatively larger pieces of food than the large folivorous monkeys previously studied, including the gorilla data increased the slope of the linear regression between body mass and Vb in anthropoids. Thus, the addition of the largest living primate brings the anthropoid Vb trend closer to the Vb trend of the order. Notwithstanding, there is still negative allometry in anthropoid Vb , in contrast with the isometry in strepsirrhine Vb . Future research should include species with body masses between the smaller anthropoids and gorillas by studying the Vb of large papionids and the other great apes.

Morbid Obesity as a Therapeutic Target for Heart Failure.

PURPOSE OF REVIEW: This review will define morbid obesity and relationship between morbid obesity and heart failure syndromes. It will delve into unique challenges facing patients with dual diagnoses of heart failure and morbid obesity and examine the data that obesity should be a target in the treatment of heart failure. RECENT FINDINGS: Emerging literature has indicated the safety and efficacy of surgical weight loss in patients with heart failure. Furthermore, bariatric surgery with associated weight loss has been associated with improvements in heart failure symptoms and reverse remodeling on echocardiography. In patients with advanced heart failure, bariatric surgery has led to improvement in heart failure to obviate the need for cardiac transplantation or sufficient weight loss for cardiac transplant eligibility. In heart failure patients who are morbidly obese, treatment of obesity is an effective therapeutic target with a myriad of potential benefits.

The Relationship Between Gait Symmetry and Metabolic Demand in Individuals With Unilateral Transfemoral Amputation: A Preliminary Study.

INTRODUCTION: Temporal-spatial symmetry allows for optimal metabolic economy in unimpaired human gait. The gait of individuals with unilateral transfemoral amputation is characterized by temporal-spatial asymmetries and greater metabolic energy expenditure. The objective of this study was to determine whether temporal-spatial asymmetries account for greater metabolic energy expenditure in individuals with unilateral transfemoral amputation. MATERIALS AND METHODS: The relationship between temporal-spatial gait asymmetry and metabolic economy (metabolic power normalized by walking speed) was retrospectively examined in eighteen individuals with transfemoral amputation walking at a self-selected velocity overground. Pearson's product-moment correlations were used to assess the relationship between: (1) step time symmetry and metabolic economy and (2) step length symmetry and metabolic economy. The retrospective analysis of this data was approved by the Walter Reed National Military Medical Center Institutional Review Board and all individuals provided written consent. Additional insights on this relationship are presented through a case series describing the temporal-spatial and metabolic responses of two individuals with transfemoral amputation who completed a split-belt treadmill walking test. RESULTS: For the cohort of individuals, there was no significant relationship between metabolic economy and either step time asymmetry or step length asymmetry. However, the case series showed a positive relationship between step length asymmetry and metabolic power as participants adapted to split-belt treadmill walking. CONCLUSION: There is mixed evidence for the relationship between temporal-spatial asymmetries and metabolic energy expenditure. This preliminary study may suggest optimal metabolic energy expenditure in individuals with transfemoral amputation occurs at an individualized level of symmetry and resultant deviations incur a metabolic penalty. The results of this study support the idea that addressing only temporal-spatial gait asymmetries in individuals with transfemoral amputation through rehabilitation may not improve metabolic economy. Nevertheless, future prospective research is necessary to confirm these results and implications for clinical practice.

Evaluating physiological signal salience for estimating metabolic energy cost from wearable sensors.

Body-in-the-loop optimization algorithms have the capability to automatically tune the parameters of robotic prostheses and exoskeletons to minimize the metabolic energy expenditure of the user. However, current body-in-the-loop algorithms rely on indirect calorimetry to obtain measurements of energy cost, which are noisy, sparsely sampled, time-delayed, and require wearing a respiratory mask. To improve these algorithms, the goal of this work is to predict a user's steady-state energy cost quickly and accurately using physiological signals obtained from portable, wearable sensors. In this paper, we quantified physiological signal salience to discover which signals, or groups of signals, have the best predictive capability when estimating metabolic energy cost. We collected data from 10 healthy individuals performing 6 activities (walking, incline walking, backward walking, running, cycling, and stair climbing) at various speeds or intensities. Subjects wore a suite of physiological sensors that measured breath frequency and volume, limb accelerations, lower limb EMG, heart rate, electrodermal activity, skin temperature, and oxygen saturation; indirect calorimetry was used to establish the 'ground truth' energy cost for each activity. Evaluating Pearson's correlation coefficients and single and multiple linear regression models with cross validation (leave-one- subject-out and leave-one- task-out), we found that 1) filtering the accelerations and EMG signals improved their predictive power, 2) global signals (e.g., heart rate, electrodermal activity) were more sensitive to unknown subjects than tasks, while local signals (e.g., accelerations) were more sensitive to unknown tasks than subjects, and 3) good predictive performance was obtained combining a small number of signals (4-5) from multiple sensor modalities. NEW & NOTEWORTHY In this paper, we systematically compare a large set of physiological signals collected from portable sensors and determine which sensor signals contain the most salient information for predicting steady-state metabolic energy cost, robust to unknown subjects or tasks. This information, together with the comprehensive data set that is published in conjunction with this paper, will enable researchers and clinicians across many fields to develop novel algorithms to predict energy cost from wearable sensors.

Study on the effects of RCCS simulated microgravity on the metabolism of human keratinocyte / 解放军医学杂志

Objective: To investigate the effects of RCCS simulated microgravity on the metabolism of the human keratinocyte cell line HaCaT. Methods: The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and HaCaT cells were cultured in vitro and divided randomly into simulated microgravity group (SMG) and normal gravity group (NG). The two group HaCaT cells were collected respectively after 1 d, 2 d and 3 d culture, and the samples were analyzed by LC/MS metabolomics. The differential metabolites between the SMG and NG cells were identified with partial least squares discriminant analysis ((O)PLS-DA), and the data were input into the KEGG database for the construction and functional analysis of metabolic pathways. Results: Comparing to NG cells, after 1 d culture, there were 74 different metabolites in SMG cells, among which 16 were up-regulated and 58 were down-regulated; after 2 d culture, there were 89 different metabolites, among which 15 were up-regulated and 74 down-regulated; after 3d culture, there were 100 different metabolites, of which 23 were up-regulated and 77 were down-regulated. The differentially expressed 49 metabolites (VIP>1 and P<0.05) after 3 d were set as target metabolites, within which the sphingosine, glutamate, and docosapentaenoic acid were down-regulated, and dehydrated sorbitol was up-regulated. KEGG analysis indicated that the metabolic pathways involved were amino acid metabolism, lipid metabolism, cell proliferation and apoptosis, substance transport, catabolism, and signal transduction. Conclusion: RCCS simulated microgravity may have significant impacts on keratinocyte metabolism mainly involving metabolites such as sphingolipids and glutamate as well as the related signaling pathways.

Nano-sized TiO2 (nTiO2) induces metabolic perturbations in Physarum polycephalum macroplasmodium to counter oxidative stress under dark conditions.

Nano-sized TiO2 (nTiO2) exerts an oxidative effect on cells upon exposure to solar or UV irradiation and ecotoxicity of the nTiO2 is an urgent concern. Little information is available regarding the effect of TiO2 on cells under dark conditions. Metabolomics is a unique approach to the discovery of biomarkers of nTiO2 cytotoxicity, and leads to the identification of perturbed metabolic pathways and the mechanism underlying nTiO2 toxicity. In the present study, gas chromatography mass spectrometry (GC/MS)-based metabolomics was performed to investigate the effect of nTiO2 on sensitive cells (P. polycephalum macroplasmodium) under dark conditions. According to the multivariate pattern recognition analysis, at least 60 potential metabolic biomarkers related to sugar metabolism, amino acid metabolism, nucleotide metabolism, polyamine biosynthesis, and secondary metabolites pathways were significantly perturbed by nTiO2. Notably, many metabolic biomarkers and pathways were related to anti-oxidant mechanisms in the living organism, suggesting that nTiO2 may induce oxidative stress, even under dark conditions. This speculation was further validated by the biochemical levels of reactive oxygen species (ROS), 8-hydroxy-2-deoxyguanosine (8-OHdG), and total soluble phenols (TSP). We inferred that the oxidative stress might be related to nTiO2-induced imbalance of cellular ROS. To the best of our knowledge, the present study is the first to investigate the nTiO2-induced metabolic perturbations in slime mold, provide a new perspective of the mechanism underlying nTiO2 toxicity under dark conditions, and show that metabolomics can be employed as a rapid, reliable and powerful tool to investigate the interaction among organisms, the environment, and nanomaterials.

Muscle recruitment and coordination with an ankle exoskeleton.

Exoskeletons have the potential to assist and augment human performance. Understanding how users adapt their movement and neuromuscular control in response to external assistance is important to inform the design of these devices. The aim of this research was to evaluate changes in muscle recruitment and coordination for ten unimpaired individuals walking with an ankle exoskeleton. We evaluated changes in the activity of individual muscles, cocontraction levels, and synergistic patterns of muscle coordination with increasing exoskeleton work and torque. Participants were able to selectively reduce activity of the ankle plantarflexors with increasing exoskeleton assistance. Increasing exoskeleton net work resulted in greater reductions in muscle activity than increasing exoskeleton torque. Patterns of muscle coordination were not restricted or constrained to synergistic patterns observed during unassisted walking. While three synergies could describe nearly 95% of the variance in electromyography data during unassisted walking, these same synergies could describe only 85-90% of the variance in muscle activity while walking with the exoskeleton. Synergies calculated with the exoskeleton demonstrated greater changes in synergy weights with increasing exoskeleton work versus greater changes in synergy activations with increasing exoskeleton torque. These results support the theory that unimpaired individuals do not exclusively use central pattern generators or other low-level building blocks to coordinate muscle activity, especially when learning a new task or adapting to external assistance, and demonstrate the potential for using exoskeletons to modulate muscle recruitment and coordination patterns for rehabilitation or performance.