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Objective: Transcranial magnetic resonance imaging (MRI)-guided histotripsy has been demonstrated to treat various locations in in vivo swine brain through a human skull. To ensure that the histotripsy treatment is delivered to the intended target location, accurate pre-treatment targeting is necessary. In this work, we investigate the feasibility of MR-thermometry and MR-acoustic radiation force imaging (MR-ARFI) to perform pre-treatment targeting of histotripsy in ex vivo bovine brain through a human skull., Methods: A 700 kHz, 128-element MR-compatible histotripsy array was used to generate histotripsy and tone-burst sonications. The array's electronic drivers were modified to also generate low-amplitude tone-burst sonications to perform MR-thermometry and MR-ARFI-based targeting. Twelve ex vivo bovine brains were treated with histotripsy at 35 MPa, 75 MPa and through a skull at 36 MPa. Before treating the tissue, both MR-ARFI and MR-thermometry were used to estimate the lesion location. Finally, the location of the histotripsy lesion was compared with the focus estimated by MR-thermometry and MR-ARFI., Results: MR-thermometry and MR-ARFI were able to successfully perform pre-treatment targeting of histotripsy using the modified histotripsy array driver. Histotripsy focus was estimated with mean absolute errors along the transverse/longitudinal axis of 2.06/2.95 mm and 2.13/2.51 mm for MR-ARFI and MR-thermometry, respectively. The presence of the human skull reduced the pressure at the focal region, but it did not compromise the targeting accuracy of either of the two methods with a mean absolute error of 1.10/2.91 mm and 1.29/2.91 mm for MR-ARFI and MR-thermometry, respectively., Conclusion: This study demonstrated that transcranial histotripsy pre-treatment targeting is feasible with MR-thermometry and MR-ARFI., Competing Interests: Conflict of interest Z.X., T.H., and the University of Michigan have financial and/or other relationships with HistoSonics Inc., (Copyright © 2024. Published by Elsevier Inc.)

Objective: To observe longitudinal correlations between limb anthropometry against weight, BMI and functional decline in patients with amyotrophic lateral sclerosis., Methods: A longitudinal, prospective, cohort study was undertaken. Four consecutive measurements of weight, height, triceps skinfold thickness (TSF), mid-upper arm (MUAC) and calf circumferences were collected at three-monthly intervals. Fat- and lean body mass were estimated using measurements of TSF and derivations of arm muscle area, respectively. Correlation analyses indicated associations between anthropometric assessments and functional decline (ALSFRS-R). Longitudinal changes were assessed using repeated measures analyses., Results: Data from 18 participants was analyzed. At enrollment, weight positively correlated with MUAC (n = 17, p = 0.0001), arm muscle area (n = 17, p = 0.04) and calf circumference (n = 17, p < 0.0001). The ALSFRS-R score negatively correlated with weight (n = 17, p = 0.03), MUAC (n = 18, p = 0.01), TSF (n = 18, p = 0.04), and calf circumference (n = 18, p = 0.003). Function significantly declined by a difference of 6.3 points per month (p = 0.009). A positive correlation was observed between the changes in weight and calf circumference over nine months (r = 0.70, p = 0.02, n  = 10)., Conclusion: Limb anthropometric measurements may be surrogate indicators of weight and BMI; TSF may be a practical, reliable indicator of fat mass, whilst changes in calf circumference may be alternatively used to monitor changes in nutritional status in the clinic.

Objective: To map the international methods used to measure energy expenditure of adults living with motor neuron disease (MND) and to highlight discrepancies when indicating hypermetabolism in the MND literature., Background: A decline in the nutritional status of patients is associated with exacerbated weight loss and shortened survival. Assessments of energy expenditure, using a variety of methods, are important to ensure an adequate energy intake to prevent malnutrition-associated weight loss. Assessments of energy expenditure are also commonly used to indicate hypermetabolism in MND, although these approaches may not be optimal., Methods: A protocol based on the Preferred Reporting Items for Systematic Reviews and Meta-analyses extension for Scoping Reviews Guidelines was developed. Three electronic databases (Medline [Ovid], CINAHL [EBSCO], and Web of Science) were exhaustively searched. Identified publications were systematically screened according to predefined PICOS eligibility criteria. The primary outcome was the identification of methods used to measure energy expenditure in MND. The secondary outcome was the identification of applications of energy expenditure assessments to indicate hypermetabolism in MND., Results: Thirty-two observational primary research publications were identified. Thirteen (40.6%) were longitudinal in design, with data on repeated measurements of energy expenditure presented in 3 (9.4%). Thirteen (40.6%) were case-control studies, of which 11 use a matched control group. Pulmonary function was used to assess eligibility in 10 publications. Energy expenditure was measured using indirect calorimetry (IC) in 31 studies. Discrepancies in the durations of fasted, measurement, and washout periods were observed. Of all included publications, 50% used assessments of resting energy expenditure to identify hypermetabolism. Bioelectrical impedance analysis was used to assess body composition alongside energy expenditure in 93.8% of publications., Conclusions: Resting energy expenditure is most frequently measured using an open-circuit IC system. However, there is a lack of a standardized, validated protocol for the conduct and reporting of IC and metabolic status in patients with MND., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Life Sciences Institute.)

Purpose: We determined the effects of shorter affected side (AS) crank arm lengths and cycling with two different prostheses on joint and crank power, asymmetry, and net efficiency., Methods: Twelve participants with a TTA rode at 1.5 W·kg -1 with equal (175 mm) and shorter AS crank arms (160, 165, 170 mm) using a daily-use prosthesis and CSP. We used statistical parametric mapping to determine differences in instantaneous joint and crank power between prostheses and linear mixed-effects models to compare average joint and crank power, asymmetry, and net efficiency., Results: Shorter AS crank arm lengths reduced the magnitude of peak positive ( P ≤ 0.001) and negative ( P < 0.001) crank power on the AS. Use of a CSP increased the magnitude of peak positive knee power ( P < 0.001) and decreased the magnitude of peak negative crank power ( P < 0.001) on the AS compared with a daily-use prosthesis. Shorter AS crank arm lengths while using a CSP reduced average hip joint ( P = 0.014) and hip transfer ( P = 0.025) power asymmetry from 35% to 20% and 118% to 62%, respectively. However, we found no significant differences in AS average joint or crank power, knee joint or crank power asymmetry, or net efficiency., Conclusions: Cycling at 1.5 W·kg -1 with unequal crank arm lengths and CSP improves hip joint power and hip transfer power asymmetry but does not alter crank asymmetry or net efficiency for recreational cyclists with a TTA., (Copyright © 2024 by the American College of Sports Medicine.)

Background: Atherosclerotic plaques form unevenly due to disturbed blood flow, causing localized endothelial cell (EC) dysfunction. Obesity exacerbates this process, but the underlying molecular mechanisms are unclear. The transcription factor EPAS1 (HIF2A) has regulatory roles in endothelium, but its involvement in atherosclerosis remains unexplored. This study investigates the potential interplay between EPAS1, obesity, and atherosclerosis., Methods: Responses to shear stress were analyzed using cultured porcine aortic EC exposed to flow in vitro coupled with metabolic and molecular analyses and by en face immunostaining of murine aortic EC exposed to disturbed flow in vivo. Obesity and dyslipidemia were induced in mice via exposure to a high-fat diet or through Leptin gene deletion. The role of Epas1 in atherosclerosis was evaluated by inducible endothelial Epas1 deletion, followed by hypercholesterolemia induction (adeno-associated virus-PCSK9 [proprotein convertase subtilisin/kexin type 9]; high-fat diet)., Results: En face staining revealed EPAS1 enrichment at sites of disturbed blood flow that are prone to atherosclerosis initiation. Obese mice exhibited substantial reduction in endothelial EPAS1 expression. Sulforaphane, a compound with known atheroprotective effects, restored EPAS1 expression and concurrently reduced plasma triglyceride levels in obese mice. Consistently, triglyceride derivatives (free fatty acids) suppressed EPAS1 in cultured EC by upregulating the negative regulator PHD2. Clinical observations revealed that reduced serum EPAS1 correlated with increased endothelial PHD2 and PHD3 in obese individuals. Functionally, endothelial EPAS1 deletion increased lesion formation in hypercholesterolemic mice, indicating an atheroprotective function. Mechanistic insights revealed that EPAS1 protects arteries by maintaining endothelial proliferation by positively regulating the expression of the fatty acid-handling molecules CD36 (cluster of differentiation 36) and LIPG (endothelial type lipase G) to increase fatty acid beta-oxidation., Conclusions: Endothelial EPAS1 attenuates atherosclerosis at sites of disturbed flow by maintaining EC proliferation via fatty acid uptake and metabolism. This endothelial repair pathway is inhibited in obesity, suggesting a novel triglyceride-PHD2 modulation pathway suppressing EPAS1 expression. These findings have implications for therapeutic strategies addressing vascular dysfunction in obesity., Competing Interests: None.

Ubiquilins are a family of cytosolic proteins that ferry ubiquitinated substrates to the proteasome for degradation. Recent work has demonstrated that Ubiquilins can also act as molecular chaperones, utilizing internal Sti1 domains to directly bind to hydrophobic sequences. Ubiquilins are associated with several neurodegenerative diseases with point mutations in UBQLN2 causing dominant, X-linked Amyotrophic Lateral Sclerosis (ALS). The molecular basis of Ubiquilin chaperone activity and how ALS mutations in the Sti1 domains affect Ubiquilin activity are poorly understood. This study presents the first crystal structure of the Sti1 domain from a fungal Ubiquilin homolog bound to a transmembrane domain (TMD). The structure reveals that two Sti1 domains form a head-to-head dimer, creating a hydrophobic cavity that accommodates two TMDs. Mapping the UBQLN2 sequence onto the structure shows that several ALS mutations are predicted to disrupt the hydrophobic groove. Using a newly developed competitive binding assay, we show that Ubiquilins preferentially bind to hydrophobic substrates with low helical propensity, motifs that are enriched in both substrates and in Ubiquilins. This study provides insights into the molecular and structural basis for Ubiquilin substrate binding, with broad implications for the role of the Sti1 domain in phase separation and ALS., Competing Interests: Competing Interests: The authors declare no competing interests.

Mitochondrial dysfunction is a common feature of C9orf72 amyotrophic lateral sclerosis/frontotemporal dementia (ALS/FTD); however, it remains unclear whether this is a cause or consequence of the pathogenic process. Analysing multiple aspects of mitochondrial biology across several Drosophila models of C9orf72 -ALS/FTD, we found morphology, oxidative stress, and mitophagy are commonly affected, which correlated with progressive loss of locomotor performance. Notably, only genetic manipulations that reversed the oxidative stress levels were also able to rescue C9orf72 locomotor deficits, supporting a causative link between mitochondrial dysfunction, oxidative stress, and behavioural phenotypes. Targeting the key antioxidant Keap1/Nrf2 pathway, we found that genetic reduction of Keap1 or pharmacological inhibition by dimethyl fumarate significantly rescued the C9orf72 -related oxidative stress and motor deficits. Finally, mitochondrial ROS levels were also elevated in C9orf72 patient-derived iNeurons and were effectively suppressed by dimethyl fumarate treatment. These results indicate that mitochondrial oxidative stress is an important mechanistic contributor to C9orf72 pathogenesis, affecting multiple aspects of mitochondrial function and turnover. Targeting the Keap1/Nrf2 signalling pathway to combat oxidative stress represents a therapeutic strategy for C9orf72 -related ALS/FTD., (© 2024 Au et al.)

Background: Although individuals with anterior cruciate ligament reconstruction (ACLR) are at high risk for posttraumatic osteoarthritis, mechanisms underlying the relationship between running and knee cartilage health remain unclear., Objective: We aimed to investigate how 30 min of running influences femoral cartilage thickness and composition and their relationships with running biomechanics in patients with ACLR and controls., Methods: Twenty patients with ACLR (time post-ACLR: 14.6 ± 6.1 months) and 20 matched controls participated in the study. A running session required both groups to run for 30 min at a self-selected speed. Before and after running, we measured femoral cartilage thickness via ultrasound imaging. A MRI session consisted of T2 mapping., Results: The ACLR group showed longer T2 relaxation times in the medial femoral condyle at resting compared with the control group (central: 51.2 ± 16.6 vs. 34.9 ± 13.2 ms, p = 0.006; posterior: 50.2 ± 10.1 vs. 39.8 ± 7.4 ms, p = 0.006). Following the run, the ACLR group showed greater deformation in the medial femoral cartilage than the control group (0.03 ± 0.01 vs. 0.01 ± 0.01 cm, p = 0.001). Additionally, the ACLR group showed significant negative correlations between resting T2 relaxation time in the medial femoral condyle and vertical impulse (standardized regression coefficients = -0.99 and p = 0.004) during running., Conclusions: Our findings suggest that those who are between 6 and 24 months post-ACLR have degraded cartilage composition and their cartilage deforms more due to running vGRF., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Passive, full-leg exoskeletons that act in parallel with the legs can reduce the metabolic power of bouncing gaits like hopping. However, the magnitude of metabolic power reduction depends on the spring stiffness profile of the exoskeleton and is presumably affected by how users adapt their lower-limb joint mechanics. We determined the effects of using a passive, full-leg exoskeleton with degressive (DG), linear (LN) and progressive (PG) stiffness springs on lower-limb joint kinematics and kinetics during stationary, bilateral hopping at 2.4 Hz. We found that the use of a passive, full-leg exoskeleton primarily reduced the muscle-tendon units (MTUs) contribution to overall joint moment and power at the ankle, followed by the knee, due to the average exoskeleton moment arm around each joint. The greatest reductions occurred with DG springs, followed by LN and PG stiffness springs, probably due to differences in elastic energy return. Moreover, the relative distribution of positive joint power remained unchanged when using a passive, full-leg exoskeleton compared with unassisted hopping. Passive, full-leg exoskeletons simultaneously assist multiple lower-limb joints and future assistive devices should consider the effects of spring stiffness profile in their design., Competing Interests: We declare we have no competing interests., (© 2024 The Authors.)

Amyotrophic lateral sclerosis (ALS) is a heterogeneous progressive neurodegenerative disorder with available treatments such as riluzole and edaravone extending survival by an average of 3-6 months. The lack of highly effective, widely available therapies reflects the complexity of ALS. Omics technologies, including genomics, transcriptomic and proteomics have contributed to the identification of biological pathways dysregulated and targeted by therapeutic strategies in preclinical and clinical trials. Integrating clinical, environmental and neuroimaging information with omics data and applying a systems biology approach can further improve our understanding of the disease with the potential to stratify patients and provide more personalised medicine. This chapter will review the omics technologies that contribute to a systems biology approach and how these components have assisted in identifying therapeutic targets. Current strategies, including the use of genetic screening and biosampling in clinical trials, as well as the future application of additional technological advances, will also be discussed., (Copyright © 2024. Published by Elsevier Inc.)

Objective: A coupling bath of circulating, chilled, degassed water is essential to safe and precise acoustic transmittance during transcranial magnetic resonance-guided focused ultrasound (tMRgFUS) procedures, but the circulating water impairs the critical real-time magnetic resonance imaging (MRI). An iron-based coupling medium (IBCM) using iron oxide nanoparticles previously developed by our group increased the relaxivity of the coupling bath such that it appears to be invisible on MRI compared with degassed water. However, the nanoparticles also reduced the pressure threshold for cavitation. To address this concern for prefocal cavitation, our group recently developed an IBCM of electrosterically stabilized and aggregation-resistant poly(methacrylic acid)-coated iron oxide nanoparticles (PMAA-FeOX) with a similar capability to reduce the MR signal of degassed water. This study examines the effect of the PMAA-FeOX IBCM on the cavitation threshold., Methods: Increasing concentrations of PMAA-FeOX nanoparticles in degassed, deionized water were placed at the focus of two different transducers to assess low and high duty-cycle pulsing parameters which are representative of two modes of focused ultrasound being investigated for tMRgFUS. Passive cavitation detection and high-speed optical imaging were used to measure cavitation threshold pressures., Results: The mean cavitation threshold was determined in both cases to be indistinguishable from the degassed water control, between 6-8 MPa for high duty-cycle pulsing (CW) and between 25.5-26.5 MPa for very low duty-cycle pulsing., Conclusion: The findings of this study indicate that an IBCM of PMAA-FeOX nanoparticles is a possible solution to reducing MRI interference from the coupling bath without increasing the risk of prefocal cavitation., Competing Interests: Conflict of Interest E.V. has research and financial relationships with HistoSonics Inc., (Copyright © 2023 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Gadolinium-based contrast agents (GBCAs) are in widespread use to enhance magnetic resonance imaging for evaluating vascular pathology. However, safety concerns and limitations regarding the use of GBCAs has led to an increased interest in alternative contrast agents. Previously, methemoglobin (metHb) and oxygen-free hemoglobin (HHb) have been shown to increase the T1-weighted signal intensity of blood, which is associated with a decrease in the T1 parameter and an enhanced contrast of the image. Thus, a lower T1 value compared to the baseline value is favorable for imaging. However, it is unknown as to whether metHb or HHb would be a stronger and more appropriate contrast agent and to what extent the T1-weighted signal is affected by concentration. This study evaluated T1-weighted images of blood samples over a range of metHb and HHb concentrations, as well as ferrous nitrosyl hemoglobin (Hb II NO) concentrations. Comparison of T1 values from a baseline value of ~ 1500 ms showed that metHb is the strongest contrast agent (T1 ~ 950 ms at 20% metHb) and that HHb is a relatively weak contrast agent (T1 ~ 1450 ms at 20% HHb). This study showed for the first time that Hb II NO can provide a contrast effect, although not as strong as metHb but stronger than HHb (T1 estimated as 1250 ms at 20% Hb II NO). With metHb providing a viable contrast between 10 and 20%, metHb has the potential to be a safe and effective contrast agent since it can be naturally converted back to hemoglobin., (© 2023. The Author(s) under exclusive licence to Biomedical Engineering Society.)

A p.Y374X truncation in TARDBP was recently shown to reduce expression of TDP43 in fibroblasts isolated from ALS cases. In this follow up study focused on assessing the downstream phenotypic consequences of loss of TDP43 in the context of the truncation, we have shown a striking effect on the fibroblast metabolic profile. Phenotypic metabolic screening uncovered a distinct metabolic profile in TDP43-Y374X fibroblasts compared to controls, which was driven by alterations in key metabolic checkpoint intermediates including pyruvate, alpha-ketoglutarate and succinate. These metabolic alterations were confirmed using transcriptomics and bioenergetic flux analysis. These data suggest that TDP43 truncation directly compromises glycolytic and mitochondrial function, identifying potential therapeutic targets for mitigating the effects of TDP43-Y374X truncation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Allen, Al Sultan, Kabucho Kibirige, Tonkiss, Hamer, Castelli, Lin, Roscoe, Stefanidis, Mead, Highley, Cooper-Knock, Hautbergue, Heath, Kirby and Shaw.)

Objective: The potential of transcranial magnetic resonance (MR)-guided histotripsy for brain applications has been described in prior in vivo studies in the swine brain through an excised human skull. The safety and accuracy of transcranial MR-guided histotripsy (tcMRgHt) rely on pre-treatment targeting guidance. In the work described here, we investigated the feasibility and accuracy of using ultrasound-induced low-temperature heating and MR thermometry for histotripsy pre-treatment targeting in ex vivo bovine brain., Methods: A 15-element, 750-kHz MRI-compatible ultrasound transducer with modified drivers that can deliver both low-temperature heating and histotripsy acoustic pulses was used to treat seven bovine brain samples. The samples were first heated to an approximately 1.6°C temperature increase at the focus, and MR thermometry was used to localize the target. Once the targeting was confirmed, a histotripsy lesion was generated at the focus and visualized on post-histotripsy MR images., Discussion: The accuracy of MR thermometry targeting was evaluated with the mean/standard deviation of the difference between the locus of peak heating identified by MR thermometry and the center of mass of the post-treatment histotripsy lesion, which was 0.59/0.31 mm and 1.31/0.93 mm in the transverse and longitudinal directions, respectively., Conclusion: This study determined that MR thermometry could provide reliable pre-treatment targeting for transcranial MR-guided histotripsy treatment., Competing Interests: Conflict of interest Z.X. and T.L.H. have financial and/or other relationships with HistoSonics Inc., (Copyright © 2022 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

We describe an autosomal dominant, multi-generational, amyotrophic lateral sclerosis (ALS) pedigree in which disease co-segregates with a heterozygous p.Y374X nonsense mutation within TDP-43. Mislocalization of TDP-43 and formation of insoluble TDP-43-positive neuronal cytoplasmic inclusions is the hallmark pathology in >95% of ALS patients. Neuropathological examination of the single case for which CNS tissue was available indicated typical TDP-43 pathology within lower motor neurons, but classical TDP-43-positive inclusions were absent from motor cortex. The mutated allele is transcribed and translated in patient fibroblasts and motor cortex tissue, but overall TDP-43 protein expression is reduced compared to wild-type controls. Despite absence of TDP-43-positive inclusions we confirmed deficient TDP-43 splicing function within motor cortex tissue. Furthermore, urea fractionation and mass spectrometry of motor cortex tissue carrying the mutation revealed atypical TDP-43 protein species but not typical C-terminal fragments. We conclude that the p.Y374X mutation underpins a monogenic, fully penetrant form of ALS. Reduced expression of TDP-43 combined with atypical TDP-43 protein species and absent C-terminal fragments extends the molecular phenotypes associated with TDP-43 mutations and with ALS more broadly. Future work will need to include the findings from this pedigree in dissecting the mechanisms of TDP-43-mediated toxicity., (© 2022 The Authors. Brain Pathology published by John Wiley & Sons Ltd on behalf of International Society of Neuropathology.)

Purpose: In this study, we examine the effects of a recently developed, iron-based coupling medium (IBCM) on guidance magnetic resonance (MR) scans during transcranial, magnetic-resonance-guided, focused ultrasound surgery (tMRgFUS) procedures. More specifically, this study tests the hypotheses that the use of the IBCM will (a) not adversely affect image quality, (b) remove aliasing from small field-of-view scans, and (c) decouple image quality from the motion state of the coupling fluid., Methods: An IBCM, whose chemical synthesis and characterization are reported elsewhere, was used as a coupling medium during tMRgFUS procedures on gel phantoms. Guidance magnetization-prepared rapid-gradient-echo (MP-RAGE), TSE, and GRE scans were acquired with fields of view of 28 and 18 cm. Experiments were repeated with the IBCM in several distinct flow states. GRE scans were used to estimate temperature time courses as a gel target was insonated. IBCM performance was measured by computing (i) the root mean square difference (RMSD) of TSE and GRE pixel values acquired using water and the IBCM, relative to the use of water; (ii) through-time temperature uncertainty for GRE scans; and (iii) Bland-Altman analysis of the temperature time courses. Finally, guidance TSE and GRE scans of a human volunteer were acquired during a separate sham tMRgFUS procedure. As a control, all experiments were repeated using a water coupling medium., Results: Use of the IBCM reduced RMSD in TSE scans by a factor of 4 or more for all fields of view and nonstationary motion states, but did not reduce RMSD estimates in MP-RAGE scans. With the coupling media in a stationary state, the IBCM altered estimates of temperature uncertainty relative to the use of water by less than 0.2°C. However, with a high flow state, the IBCM reduced temperature uncertainties by the statistically significant amounts (at the 0.01 level) of 0.5°C (28 cm field of view) and 5°C (18 cm field of view). Bland-Altman analyses found a 0.1°C ± 0.5°C difference between temperature estimates acquired using water and the IBCM as coupling media. Finally, scans of a human volunteer using the IBCM indicate more conspicuous grey/white matter contrast, a reduction in aliasing, and a less than 0.2°C change in temperature uncertainty., Conclusions: The use of an IBCM during tMRgFUS procedures does not adversely affect image quality for TSE and GRE scans, can decouple image quality from the motion state of the coupling fluid, and can remove aliasing from scans where the field of view is set to be much smaller than the spatial extent of the coupling fluid., (© 2022 American Association of Physicists in Medicine.)

The volume of active muscle and duration of extensor muscle force well explain the associated metabolic energy expenditure across body mass and velocity during level-ground running and hopping. However, if these parameters fundamentally drive metabolic energy expenditure, then they should pertain to multiple modes of locomotion and provide a simple framework for relating biomechanics to metabolic energy expenditure in bouncing gaits. Therefore, we evaluated the ability of the 'cost of generating force' hypothesis to link biomechanics and metabolic energy expenditure during human running and hopping across step frequencies. We asked participants to run and hop at 85%, 92%, 100%, 108% and 115% of preferred running step frequency. We calculated changes in active muscle volume, duration of force production and metabolic energy expenditure. Overall, as step frequency increased, active muscle volume decreased as a result of postural changes via effective mechanical advantage (EMA) or duty factor. Accounting for changes in EMA and muscle volume better related to metabolic energy expenditure during running and hopping at different step frequencies than assuming a constant EMA and muscle volume. Thus, to ultimately develop muscle mechanics models that can explain metabolic energy expenditure across different modes of locomotion, we suggest more precise measures of muscle force production that include the effects of EMA., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Purpose: To develop a new approach to 2D turbo spin -echo (TSE) imaging using annular spiral rings with a retraced in/out trajectory, dubbed "SPRING-RIO TSE", for fast T 2 -weighted brain imaging at 3T., Methods: A long spiral trajectory was split into annular segmentations that were then incorporated into a 2D TSE acquisition module to fully exploit the sampling efficiency of spiral rings. A retraced in/out trajectory strategy coupled with spiral-ring TSE was introduced to increase SNR, mitigate T 2 -decay induced artifacts, and self-correct moderate off-resonance while maintaining the target TE and causing no scan time penalty. Model-based k-space estimation and semiautomatic off-resonance correction algorithms were implemented to minimize effects of k-space trajectory infidelity and B 0 inhomogeneity, respectively. The resulting SPRING-RIO TSE method was compared to the original spiral-ring (abbreviated "SPRING") TSE and Cartesian TSE using simulations, and phantom and in vivo acquisitions., Results: Simulation and phantom studies demonstrated the performance of the proposed SPRING-RIO TSE pulses sequence, as well as that of trajectory correction and off-resonance correction. Volunteer data showed that the proposed method achieves high-quality 2D T 2 -weighted brain imaging with a higher scan efficiency (0:45 min/14 slices versus 1:31 min/14 slices), improved image contrast, and reduced specific absorption rate compared to conventional 2D Cartesian TSE., Conclusion: 2D T 2 -weighted brain imaging using spiral-ring TSE was implemented and tested, providing several potential advantages over conventional 2D Cartesian TSE imaging., (© 2022 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals LLC on behalf of International Society for Magnetic Resonance in Medicine.)

Amyotrophic lateral sclerosis is a rapidly progressive neurodegenerative disease that affects 1/350 individuals in the United Kingdom. The cause of amyotrophic lateral sclerosis is unknown in the majority of cases. Two-sample Mendelian randomization enables causal inference between an exposure, such as the serum concentration of a specific metabolite, and disease risk. We obtained genome-wide association study summary statistics for serum concentrations of 566 metabolites which were population matched with a genome-wide association study of amyotrophic lateral sclerosis. For each metabolite, we performed Mendelian randomization using an inverse variance weighted estimate for significance testing. After stringent Bonferroni multiple testing correction, our unbiased screen revealed three metabolites that were significantly linked to the risk of amyotrophic lateral sclerosis: Estrone-3-sulphate and bradykinin were protective, which is consistent with literature describing a male preponderance of amyotrophic lateral sclerosis and a preventive effect of angiotensin-converting enzyme inhibitors which inhibit the breakdown of bradykinin. Serum isoleucine was positively associated with amyotrophic lateral sclerosis risk. All three metabolites were supported by robust Mendelian randomization measures and sensitivity analyses; estrone-3-sulphate and isoleucine were confirmed in a validation amyotrophic lateral sclerosis genome-wide association study. Estrone-3-sulphate is metabolized to the more active estradiol by the enzyme 17β-hydroxysteroid dehydrogenase 1; further, Mendelian randomization demonstrated a protective effect of estradiol and rare variant analysis showed that missense variants within HSD17B1 , the gene encoding 17β-hydroxysteroid dehydrogenase 1, modify risk for amyotrophic lateral sclerosis. Finally, in a zebrafish model of C9ORF72 -amyotrophic lateral sclerosis, we present evidence that estradiol is neuroprotective. Isoleucine is metabolized via methylmalonyl-CoA mutase encoded by the gene MMUT in a reaction that consumes vitamin B12. Multivariable Mendelian randomization revealed that the toxic effect of isoleucine is dependent on the depletion of vitamin B12; consistent with this, rare variants which reduce the function of MMUT are protective against amyotrophic lateral sclerosis. We propose that amyotrophic lateral sclerosis patients and family members with high serum isoleucine levels should be offered supplementation with vitamin B12., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Guarantors of Brain.)

Adenosine deaminase (ADA) is a purine metabolism enzyme that catalyses the breakdown of adenosine and deoxyadenosine. The enzyme is important in several cellular processes, including the innate immune response and cellular differentiation, and it is also an important enzyme for the maintenance of brain homeostasis, in part due to its regulation of adenosine. Aberrant regulation of ADA enzyme activity has been linked to several neurodegenerative diseases and diseases that can result in neurological impairment. However, the mechanisms behind altered ADA regulation and how this leads to the development of neurological dysfunction are poorly characterised. This review summarises the current research on ADA and its role and regulation in disease pathology, with a focus on the central nervous system (CNS) and the neurodegenerative disease, amyotrophic lateral sclerosis (ALS).

All stage resistance to stripe rust races prevalent in India was investigated in the European winter wheat cultivar 'Acienda'. In order to dissect the genetic basis of the resistance, a backcross population was developed between 'Acienda' and the stripe rust susceptible Indian spring wheat cultivar 'HD 2967'. Inheritance studies revealed segregation for a dominant resistant gene. High density SNP genotyping was used to map stripe rust resistance and marker regression analysis located stripe rust resistance to the distal end of wheat chromosome 1A. Interval mapping located this region between the SNP markers AX-95162217 and AX-94540853, at a LOD score of 15.83 with a phenotypic contribution of 60%. This major stripe rust resistance locus from 'Acienda' has been temporarily designated as Yraci. A candidate gene search in the 2.76 Mb region carrying Yraci on chromosome 1A identified 18 NBS-LRR genes based on wheat RefSeqv1.0 annotations. Our results indicate that as there is no major gene reported in the Yraci chromosome region, it is likely to be a novel stripe rust resistance locus and offers potential for deployment, using the identified markers, to confer all stage stripe rust resistance., Competing Interests: The authors have declared that no competing interests exist.

Purpose: Intrinsic foot muscles maintain foot structural integrity and contribute to functional movement, posture and balance. Thus, assessing intrinsic foot muscle size and strength are important. Magnetic resonance imaging (MRI) has been shown to accurately image the individual muscles but is costly and time consuming. Ultrasound (US) imaging may provide an alternative that is less costly and more readily available. The purpose of this study was to investigate the validity and intratester reliability of US imaging in measuring intrinsic foot muscle size in comparison to MRI., Methods: US and MRI were employed to measure the intrinsic foot muscle size involving 35 participants (females = 13; males = 22). The scanned intrinsic foot muscles included the flexor hallucis brevis (FHB), abductor hallucis (ABDH), flexor digitorum brevis (FDB), quadratus plantae (QP) and abductor digiti minimi (ADM). Pearson product correlation (r), intraclass correlation coefficients (ICC), standard error of the measurement (SEm) and minimal detectable difference (MDD) were calculated., Results: High correlations were detected between the US and MRI cross-sectional area (CSA) measurements (r = .971 to 0.995). Test reliability was excellent for both MRI and US (ICC = 0.994 to 0.999). Limits of agreement between MRI and US measurements from ranged from 5.7 to 12.2% of muscle size. SEm values for US ranged from 0.026 to 0.044 cm2, while the SEm for MRI ranged from 0.018 to 0.023 cm2. MDD values for US ranged from 0.073 to 0.122 cm2, while MRI ranged from 0.045 to 0.064 cm2., Conclusions: US appears to be a valid and reliable alternative to MRI when measuring intrinsic foot muscle CSA. While US is less costly and more readily available, the MRI results were shown to be slightly more precise., (© 2022. The Author(s).)

Mechanical cues play a vital role in limb skeletal development, yet their influence and underpinning mechanisms in the regulation of endochondral ossification (EO) processes are incompletely defined. Furthermore, interactions between endochondral growth and mechanics and the mTOR/NF-ĸB pathways are yet to be explored. An appreciation of how mechanical cues regulate EO would also clearly be beneficial in the context of fracture healing and bone diseases, where these processes are recapitulated. The study herein addresses the hypothesis that the mTOR/NF-ĸB pathways interact with mechanics to control endochondral growth. To test this, murine embryonic metatarsals were incubated ex vivo in a hydrogel, allowing for the effects of quasi-static loading on longitudinal growth to be assessed. The results showed significant restriction of metatarsal growth under quasi-static loading during a 14-day period and concentration-dependent sensitivity to hydrogel-related restriction. This study also showed that hydrogel-treated metatarsals retain their viability and do not present with increased apoptosis. Metatarsals exhibited reversal of the growth-restriction when co-incubated with mTOR compounds, whilst it was found that these compounds showed no effects under basal culture conditions. Transcriptional changes linked to endochondral growth were assessed and downregulation of Col2 and Acan was observed in hydrogel-treated metatarsi at day 7. Furthermore, cell cycle analyses confirmed the presence of chondrocytes exhibiting S-G2/M arrest. These data indicate that quasi-static load provokes chondrocyte cell cycle arrest, which is partly overcome by mTOR, with a less marked interaction for NF-ĸB regulators.

Aging is a major risk factor for neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). As metabolic alterations are a hallmark of aging and have previously been observed in ALS, it is important to examine the effect of aging in the context of ALS metabolic function. Here, using a newly established phenotypic metabolic approach, we examined the effect of aging on the metabolic profile of fibroblasts derived from ALS cases compared to controls. We found that ALS fibroblasts have an altered metabolic profile, which is influenced by age. In control cases, we found significant increases with age in NADH metabolism in the presence of several metabolites including lactic acid, trehalose, uridine and fructose, which was not recapitulated in ALS cases. Conversely, we found a reduction of NADH metabolism with age of biopsy, age of onset and age of death in the presence of glycogen in the ALS cohort. Furthermore, we found that NADH production correlated with disease progression rates in relation to a number of metabolites including inosine and α-ketoglutaric acid. Inosine or α-ketoglutaric acid supplementation in ALS fibroblasts was bioenergetically favourable. Overall, we found aging related defects in energy substrates that feed carbon into glycolysis at various points as well as the tricarboxylic acid (TCA) cycle in ALS fibroblasts, which was validated in induced neuronal progenitor cell derived iAstrocytes. Our results suggest that supplementing those pathways may protect against age related metabolic dysfunction in ALS., Competing Interests: Conflict of interest The authors can state there is no conflict of interest associated with this work., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Astrocytes are the most abundant glial cell in the central nervous system and are involved in multiple processes including metabolic homeostasis, blood brain barrier regulation and neuronal crosstalk. Astrocytes are the main storage point of glycogen in the brain and it is well established that astrocyte uptake of glutamate and release of lactate prevents neuronal excitability and supports neuronal metabolic function. However, the role of lipid metabolism in astrocytes in relation to neuronal support has been until recently, unclear. Lipids play a fundamental role in astrocyte function, including energy generation, membrane fluidity and cell to cell signaling. There is now emerging evidence that astrocyte storage of lipids in droplets has a crucial physiological and protective role in the central nervous system. This pathway links β-oxidation in astrocytes to inflammation, signalling, oxidative stress and mitochondrial energy generation in neurons. Disruption in lipid metabolism, structure and signalling in astrocytes can lead to pathogenic mechanisms associated with a range of neurological disorders., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Purpose: Intraoperative T 2 -weighted (T2-w) imaging unreliably captures image contrast specific to thermal ablation after transcranial MR-guided focused ultrasound surgery, impeding dynamic imaging feedback. Using a porcine thalamotomy model, we test the unproven hypothesis that intraoperative DWI can improve dynamic feedback by detecting lesioning within 30 minutes of transcranial MR-guided focused ultrasound surgery., Methods: Twenty-five thermal lesions were formed in six porcine models using a clinical transcranial MR-guided focused ultrasound surgery system. A novel diffusion-weighted pulse sequence monitored the formation of T2-w and diffusion-weighted lesion contrast after ablation. Using postoperative T2-w contrast to indicate lesioning, apparent intraoperative image contrasts and diffusion coefficients at each lesion site were computed as a function of time after ablation, observed peak temperature, and observed thermal dose. Lesion sizes segmented from imaging and thermometry were compared. Image reviewers estimated the time to emergence of lesion contrast. Intraoperative image contrasts were analyzed using receiver operator curves., Results: On average, the apparent diffusion coefficient at lesioned sites decreased within 5 minutes after ablation relative to control sites. In-plane lesion areas on intraoperative DWI varied from postoperative T2-w MRI and MR thermometry by 9.6 ± 9.7 mm 2 and - 4.0 ± 7.1 mm 2 , respectively. The 0.25, 0.5, and 0.75 quantiles of the earliest times of observed T2-w and diffusion-weighted lesion contrast were 10.7, 21.0, and 27.8 minutes and 3.7, 8.6, and 11.8 minutes, respectively. The T2-w and diffusion-weighted contrasts and apparent diffusion coefficient values produced areas under the receiver operator curve of 0.66, 0.80, and 0.74, respectively., Conclusion: Intraoperative DWI can detect MR-guided focused ultrasound surgery lesion formation in the brain within several minutes after treatment., (© 2020 International Society for Magnetic Resonance in Medicine.)

Neurodegenerative diseases are a group of nervous system conditions characterised pathologically by the abnormal deposition of protein throughout the brain and spinal cord. One common pathophysiological change seen in all neurodegenerative disease is a change to the metabolic function of nervous system and peripheral cells. Glycolysis is the conversion of glucose to pyruvate or lactate which results in the generation of ATP and has been shown to be abnormal in peripheral cells in Alzheimer's disease, Parkinson's disease, and Amyotrophic Lateral Sclerosis. Changes to the glycolytic pathway are seen early in neurodegenerative disease and highlight how in multiple neurodegenerative conditions pathology is not always confined to the nervous system. In this paper, we review the abnormalities described in glycolysis in the three most common neurodegenerative diseases. We show that in all three diseases glycolytic changes are seen in fibroblasts, and red blood cells, and that liver, kidney, muscle and white blood cells have abnormal glycolysis in certain diseases. We highlight there is potential for peripheral glycolysis to be developed into multiple types of disease biomarker, but large-scale bio sampling and deciphering how glycolysis is inherently altered in neurodegenerative disease in multiple patients' needs to be accomplished first to meet this aim.

Hypoxia is a feature of neurodegenerative diseases, and can both directly and indirectly impact on neuronal function through modulation of glial function. Astrocytes play a key role in regulating homeostasis within the central nervous system, and mediate hypoxia-induced changes in response to reduced oxygen availability. The current study performed a detailed characterization of hypoxia-induced changes in the transcriptomic profile of astrocytes in vitro. Human astrocytes were cultured under normoxic (5% CO 2 , 95% air) or hypoxic conditions (1% O 2 , 5% CO 2 , 94% N 2 ) for 24 h, and the gene expression profile assessed by microarray analysis. In response to hypoxia 4904 genes were significantly differentially expressed (1306 upregulated and 3598 downregulated, FC ≥ 2 and p ≤ 0.05). Analysis of the significant differentially expressed transcripts identified an increase in immune response pathways, and dysregulation of signalling pathways, including HIF-1 ( p = 0.002), and metabolism, including glycolysis ( p = 0.006). To assess whether the hypoxia-induced metabolic gene changes observed affected metabolism at a functional level, both the glycolytic and mitochondrial flux were measured using an XF bioanalyser. In support of the transcriptomic data, under physiological conditions hypoxia significantly reduced mitochondrial respiratory flux ( p = 0.0001) but increased basal glycolytic flux ( p = 0.0313). However, when metabolically stressed, hypoxia reduced mitochondrial spare respiratory capacity ( p = 0.0485) and both glycolytic capacity ( p = 0.0001) and glycolytic reserve ( p < 0.0001). In summary, the current findings detail hypoxia-induced changes in the astrocyte transcriptome in vitro, identifying potential targets for modifying the astrocyte response to reduced oxygen availability in pathological conditions associated with ischaemia/hypoxia, including manipulation of mitochondrial function, metabolism, and the immune response.

Purpose: We sought to biomechanically distinguish steep uphill running from steep uphill walking and explore why athletes alternate between walking and running on steep inclines., Methods: We quantified vertical center of mass (COM) accelerations and basic stride parameters for both walking and running at a treadmill speed of 1.0 m/s on the level and up a 30° incline. We also investigated how electromyography (EMG) of the gluteus maximus (GMAX), vastus medialis (VM), medial gastrocnemius (MG), and soleus (SOL) muscles differ between gaits when ascending steep hills., Results: The vertical COM accelerations for steep uphill walking exhibited two peaks per step of magnitude 1.47 ± 0.23 g and 0.79 ± 0.10 g. In contrast, steep running exhibited a single peak per step pattern with a magnitude of 1.81 ± 0.15 g. Steep uphill running exhibited no aerial phase, 40% faster stride frequency, and 40% shorter foot-ground contact time compared to steep uphill walking but similar leg swing times. SOL showed 36% less iEMG per stride during steep uphill running versus steep uphill walking, but all other EMG comparisons between steep running and walking were not significantly different., Conclusions: Multiple biomechanical variables clearly indicate that steep uphill running is a distinctly different gait from steep uphill walking and is more similar to level running. The competing desires to minimize the energetic cost of locomotion and to avoid exhaustion of the SOL may be a possible explanation for gait alternation on steep inclines.

Amyotrophic lateral sclerosis (ALS) is a rapidly progressive and fatal neurodegenerative disorder for which there is no effective curative treatment available and minimal palliative care. Mutations in the gene encoding the TAR DNA-binding protein 43 (TDP-43) are a well-recognized genetic cause of ALS, and an imbalance in energy homeostasis correlates closely to disease susceptibility and progression. Considering previous research supporting a plethora of downstream cellular impairments originating in the histopathological signature of TDP-43, and the solid evidence around metabolic dysfunction in ALS, a causal association between TDP-43 pathology and metabolic dysfunction cannot be ruled out. Here we discuss how TDP-43 contributes on a molecular level to these impairments in energy homeostasis, and whether the protein's pathological effects on cellular metabolism differ from those of other genetic risk factors associated with ALS such as superoxide dismutase 1 (SOD1), chromosome 9 open reading frame 72 (C9orf72) and fused in sarcoma (FUS)., Competing Interests: Declaration of conflicting interests:The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2020.)

Competing Interests: None

Purpose: To increase volume coverage in real-time MR thermometry for transcranial MR-guided focused ultrasound (tcMRgFUS) ablation, without multiple receive coils., Theory and Methods: Multiband excitation and incoherent blipped-controlled aliasing were implemented in a 2DFT pulse sequence used clinically for tcMRgFUS, and an extended k-space hybrid reconstruction was developed that recovers slice-separated temperature maps assuming that heating is focal, given slice-separated pretreatment images. Simulations were performed to characterize slice leakage, the number of slices that can be simultaneously imaged with low-temperature error, and robustness across random slice-phase k-space permutations. In vivo experiments were performed using a single receive coil without heating to measure temperature precision, and gel phantom FUS experiments were performed to test the method with heating and with a water bath., Results: Simulations showed that with large hot spots and identical magnitude images on each slice, up to three slices can be simultaneously imaged with less than 1 ∘ C temperature root-mean-square error. They also showed that hot spots do not alias coherently between slices, and that an average 86% of random slice-phase k-space permutations yielded less than 1 ∘ C temperature error. Temperature precision was not degraded compared to single-slice imaging in the in vivo SMS scans, and the gel phantom SMS temperature maps closely tracked single-slice temperature in the hot spot, with no coherent aliasing to other slices., Conclusions: Incoherent controlled aliasing SMS enables accurate reconstruction of focal heating maps from two or three slices simultaneously, using a single receive coil and a sparsity-promoting temperature reconstruction., (© 2019 International Society for Magnetic Resonance in Medicine.)

Upright standing involves small displacements of the center of mass about the base of support. These displacements are often quantified by measuring various kinematic features of the center-of-pressure trajectory. The plantar flexors have often been identified as the key muscles for the control of these displacements; however, studies have suggested that the hip abductor and adductors may also be important. The purpose of our study was to determine the association between the force capabilities of selected leg muscles and sway-area rate across four balance conditions in young (25 ± 4 years; 12/19 women) and older adults (71 ± 5 years; 5/19 women). Due to the marked overlap in sway-area rate between the two age groups, the data were collapsed, and individuals were assigned to groups of low- and high-sway area rates based on a k-medoid cluster analysis. The number of participants assigned to each group varied across balance conditions and a subset of older adults was always included in the low-sway group for each balance condition. The most consistent explanatory variable for the variance in sway-area rate was force control of the hip abductors and ankle dorsiflexors as indicated by the magnitude of the normalized force fluctuations (force steadiness) during a submaximal isometric contraction. The explanatory power of the regression models varied across conditions, thereby identifying specific balance conditions that should be examined further in future studies of postural control.

Purpose: Acoustic coupling baths, nominally composed of degassed water, play important roles during transcranial focused ultrasound surgery. However, this large water bolus also degrades the quality of intraoperative magnetic resonance (MR) guidance imaging. In this study, we test the feasibility of using dilute, aqueous magnetite nanoparticle suspensions to suppress these image degradations while preserving acoustic compatibility. We examine the effects of these suspensions on metrics of image quality and acoustic compatibility for two types of transcranial focused ultrasound insonation regimes: low-duty cycle histotripsy procedures and high-duty cycle thermal ablation procedures., Methods: Magnetic resonance guidance imaging was used to monitor thermal ablations of in vitro gel targets using a coupling bath composed of various concentrations of aqueous, suspended, magnetite nanoparticles in a clinical transcranial transducer under stationary and flowing conditions. Thermal deposition was monitored using MR thermometry simultaneous to insonation. Then, using normal degassed water as a coupling bath, various concentrations of aqueous, suspended, magnetite nanoparticles were placed at the center of this same transducer and insonated using high-duty cycle pulsing parameters. Passive cavitation detectors recorded cavitation emissions, which were then used to estimate the relative number of cavitation events per insonation (cavitation duty cycle) and the cavitation dose estimates of each nanoparticle concentration. Finally, the nanoparticle mixtures were exposed to low-duty cycle, histotripsy pulses. Passive cavitation detectors monitored cavitation emissions, which were used to estimate cavitation threshold pressures., Results: The nanoparticles reduced the MR signal of the coupling bath by 90% in T2- and T2*-weighted images and also removed almost all imaging artifacts caused by coupling bath motion. The coupling baths caused <5% changes in peak temperature change achieved during sonication, as observed via MR thermometry. At low duty cycle insonations, the nanoparticles decreased the cavitation threshold pressure by about 15 ± 7% in a manner uncorrelated with nanoparticle concentration. At high duty cycle insonations, the 0.5 cavitation duty cycle acoustic power threshold varied linearly with nanoparticle concentration., Conclusions: Dilute aqueous magnetite nanoparticle suspensions effectively reduced MR imaging artifacts caused by the acoustic coupling bath. They also attenuated acoustic power deposition by <5%. For low duty cycle insonation regimes, the nanoparticles decreased the cavitation threshold by 15 ± 7%. However, for high-duty cycle regimes, the nanoparticles decreased the threshold for cavitation in proportion to nanoparticle concentration., (© 2019 American Association of Physicists in Medicine.)

It is important to understand how the disease process affects the metabolic pathways in amyotrophic lateral sclerosis and whether these pathways can be manipulated to ameliorate disease progression. To analyse the basis of the metabolic defect in amyotrophic lateral sclerosis we used a phenotypic metabolic profiling approach. Using fibroblasts and reprogrammed induced astrocytes from C9orf72 and sporadic amyotrophic lateral sclerosis cases we measured the production rate of reduced nicotinamide adenine dinucleotides (NADH) from 91 potential energy substrates simultaneously. Our screening approach identified that C9orf72 and sporadic amyotrophic lateral sclerosis induced astrocytes have distinct metabolic profiles compared to controls and displayed a loss of metabolic flexibility that was not observed in fibroblast models. This loss of metabolic flexibility, involving defects in adenosine, fructose and glycogen metabolism, as well as disruptions in the membrane transport of mitochondrial specific energy substrates, contributed to increased starvation induced toxicity in C9orf72 induced astrocytes. A reduction in glycogen metabolism was attributed to loss of glycogen phosphorylase and phosphoglucomutase at the protein level in both C9orf72 induced astrocytes and induced neurons. In addition, we found alterations in the levels of fructose metabolism enzymes and a reduction in the methylglyoxal removal enzyme GLO1 in both C9orf72 and sporadic models of disease. Our data show that metabolic flexibility is important in the CNS in times of bioenergetic stress., (© The Author(s) (2019). Published by Oxford University Press on behalf of the Guarantors of Brain.)

Amyotrophic lateral sclerosis (ALS) is an adult onset neurological disorder characterized by loss of motor neurons leading to progressive muscle wasting and eventually death. Astrocytes play a key role in disease pathogenesis. However, the ability to study astrocytic support towards motor neurons in ALS has been limited by a lack of sustainable high-throughput human cell models. Moreover, the ability to assess how astrocytic support of motor neurons is influenced by drug treatment or nutritional supplementation has been hampered by the lack of robust methodology. We have developed a high-throughput astrocyte motor neuron co-culture assay, which, by using Hb9-GFP+ motor neurons enables researchers to assess how ALS affects the ability of astrocytes to support motor neurons in 384-well plates. Moreover, astrocyte function can be manipulated by nutritional supplementation or drug treatment to identify possible therapeutic targets., Competing Interests: Competing interests M.J.S. and L.F. are funded by the biotech company BenevolentAI.

When humans hop with a passive-elastic exoskeleton with springs in parallel with both legs, net metabolic power (P met ) decreases compared with normal hopping (NH). Furthermore, humans retain near-constant total vertical stiffness ( k tot ) when hopping with such an exoskeleton. To determine how spring stiffness profile affects P met and biomechanics, 10 subjects hopped on both legs normally and with three full-leg exoskeletons that each used a different spring stiffness profile at 2.4, 2.6, 2.8, and 3.0 Hz. Each subject hopped with an exoskeleton that had a degressive spring stiffness (DG exo ), where stiffness, the slope of force vs. displacement, is initially high but decreases with greater displacement, linear spring stiffness (LN exo ), where stiffness is constant, or progressive spring stiffness (PG exo ), where stiffness is initially low but increases with greater displacement. Compared with NH, use of the DG exo , LN exo , and PG exo numerically resulted in 13-24% lower, 4-12% lower, and 0-8% higher P met , respectively, at 2.4-3.0 Hz. Hopping with the DG exo reduced P met compared with NH at 2.4-2.6 Hz ( P ≤ 0.0457) and reduced P met compared with the PG exo at 2.4-2.8 Hz ( P < 0.001). k tot while hopping with each exoskeleton was not different compared with NH, suggesting that humans adjust leg stiffness to maintain overall stiffness regardless of the spring stiffness profile in an exoskeleton. Furthermore, the DG exo provided the greatest elastic energy return, followed by LN exo and PG exo ( P ≤ 0.001). Future full-leg, passive-elastic exoskeleton designs for hopping, and presumably running, should use a DG exo rather than an LN exo or a PG exo to minimize metabolic demand. NEW & NOTEWORTHY When humans hop at 2.4-3.0 Hz normally and with an exoskeleton with different spring stiffness profiles in parallel to the legs, net metabolic power is lowest when hopping with an exoskeleton with degressive spring stiffness. Total vertical stiffness is constant when using an exoskeleton with linear or nonlinear spring stiffness compared with normal hopping. In-parallel spring stiffness influences net metabolic power and biomechanics and should be considered when designing passive-elastic exoskeletons for hopping and running.

As clinical evidence supports a negative impact of dysfunctional energy metabolism on the disease progression in amyotrophic lateral sclerosis, it is vital to understand how the energy metabolic pathways are altered and whether they can be restored to slow disease progression. Possible approaches include increasing or rerouting catabolism of alternative fuel sources to supplement the glycolytic and mitochondrial pathways such as glycogen, ketone bodies and nucleosides. To analyse the basis of the catabolic defect in amyotrophic lateral sclerosis we used a novel phenotypic metabolic array. We profiled fibroblasts and induced neuronal progenitor-derived human induced astrocytes from C9orf72 amyotrophic lateral sclerosis patients compared to normal controls, measuring the rates of production of reduced nicotinamide adenine dinucleotides from 91 potential energy substrates. This approach shows for the first time that C9orf72 human induced astrocytes and fibroblasts have an adenosine to inosine deamination defect caused by reduction of adenosine deaminase, which is also observed in induced astrocytes from sporadic patients. Patient-derived induced astrocyte lines were more susceptible to adenosine-induced toxicity, which could be mimicked by inhibiting adenosine deaminase in control lines. Furthermore, adenosine deaminase inhibition in control induced astrocytes led to increased motor neuron toxicity in co-cultures, similar to the levels observed with patient derived induced astrocytes. Bypassing metabolically the adenosine deaminase defect by inosine supplementation was beneficial bioenergetically in vitro, increasing glycolytic energy output and leading to an increase in motor neuron survival in co-cultures with induced astrocytes. Inosine supplementation, in combination with modulation of the level of adenosine deaminase may represent a beneficial therapeutic approach to evaluate in patients with amyotrophic lateral sclerosis., (© The Author(s) (2019). Published by Oxford University Press on behalf of the Guarantors of Brain.)

Purpose: Images acquired with spiral k-space trajectories can suffer from off-resonance image blur. Previous work showed that averaging 2 images acquired with a retraced, in/out (RIO) trajectory self-corrects image blur so long as off-resonant spins accrue less than 1 half-cycle of relative phase over the readout. Practical scenarios frequently exceed this threshold. Here, we derive and characterize a more-robust off-resonance image blur correction method for RIO acquisitions., Methods: Phantom and human volunteer data were acquired using a RIO trajectory with readout durations ranging from 4 to 60 ms. The resulting images were deblurred using 3 candidate methods: conventional linear correction of the component images; semiautomatic deblurring of the component images using an established minimized phase objective function; and semiautomatic deblurring of the average of the component images using a maximized energy objective function, derived below. Deblurring errors were estimated relative to images acquired with 4 ms readouts., Results: All 3 methods converged to similar solutions in cases where less than 2 and 4 cycles of phase accrued over the readout in in vivo and phantom images, respectively (<13 ms readout at 3T). Above this threshold, the linear and minimized phase methods introduced several errors. The maximized energy function provided accurate deblurring so long as less than 6 and 10 cycles of phase accrued over the readout in in vivo and phantom images, respectively (<34 ms readout at 3T)., Conclusion: The maximized energy objective function can accurately deblur RIO acquisitions over a wide spectrum of off resonance frequencies., (© 2018 International Society for Magnetic Resonance in Medicine.)

Objective: Histotripsy is an ultrasound-based treatment modality relying on the generation of targeted cavitation bubble clouds, which mechanically fractionate tissue. The purpose of the current study was to investigate the in vivo feasibility, including dosage requirements and safety, of generating well-confined destructive lesions within the porcine brain utilizing histotripsy technology., Methods: Following a craniectomy to open an acoustic window to the brain, histotripsy pulses were delivered to generate lesions in the porcine cortex. Large lesions with a major dimension of up to 1 cm were generated to demonstrate the efficacy of histotripsy lesioning in the brain. Gyrus-confined lesions were generated at different applied dosages and under ultrasound imaging guidance to ensure that they were accurately targeted and contained within individual gyri. Clinical evaluation as well as MRI and histological outcomes were assessed in the acute (≤ 6 hours) and subacute (≤ 72 hours) phases of recovery., Results: Histotripsy was able to generate lesions with a major dimension of up to 1 cm in the cortex. Histotripsy lesions were seen to be well demarcated with sharp boundaries between treated and untreated tissues, with histological evidence of injuries extending ≤ 200 µm from their boundaries in all cases. In animals with lesions confined to the gyrus, no major hemorrhage or other complications resulting from treatment were observed. At 72 hours, MRI revealed minimal to no edema and no radiographic evidence of inflammatory changes in the perilesional area. Histological evaluation revealed the histotripsy lesions to be similar to subacute infarcts., Conclusions: Histotripsy can be used to generate sharply defined lesions of arbitrary shapes and sizes in the swine cortex. Lesions confined to within the gyri did not lead to significant hemorrhage or edema responses at the treatment site in the acute or subacute time intervals.

Equine chorionic girdle trophoblast cells play important endocrine and immune functions critical in supporting pregnancy. Very little is known about the genes and pathways that regulate chorionic girdle trophoblast development. Our aim was to identify genes and signalling pathways active in vivo in equine chorionic girdle trophoblast within a critical 7-days window. We exploited the late implantation of the equine conceptus to obtain trophoblast tissue. An Agilent equine 44K microarray was performed using RNA extracted from chorionic girdle and chorion (control) from equine pregnancy days 27, 30, 31 and 34 (n = 5), corresponding to the initiation of chorionic girdle trophoblast proliferation, differentiation and migration. Data were analysed using R packages limma and maSigPro, Ingenuity Pathway Analysis and DAVID and verified using qRT-PCR, promoter analysis, western blotting and migration assays. Microarray analysis showed gene expression (absolute log FC >2, FDR-adjusted P < 0.05) was rapidly and specifically induced in the chorionic girdle between days 27 and 34 (compared to day 27, day 30 = 116, day 31 = 317, day 34 = 781 genes). Pathway analysis identified 35 pathways modulated during chorionic girdle development (e.g. FGF, integrin, Rho GTPases, MAPK) including pathways that have limited description in mammalian trophoblast (e.g. IL-9, CD40 and CD28 signalling). Rho A and ERK/MAPK activity was confirmed as was a role for transcription factor ELF5 in regulation of the CGB promoter. The purity and accessibility of chorionic girdle trophoblast proved to be a powerful resource to identify candidate genes and pathways involved in early equine placental development., (2018 The authors)

Histotripsy is a non-invasive, non-thermal ablation technique that uses high-amplitude, focused ultrasound pulses to fractionate tissue via acoustic cavitation. The goal of this study was to illustrate the potential of histotripsy with electronic focal steering to achieve rapid ablation of a tissue volume at a rate matching or exceeding those of current clinical techniques (∼1-2 mL/min). Treatment parameters were established in tissue-mimicking phantoms and applied to ex vivo tissue. Six-microsecond pulses were delivered by a 250-kHz array. The focus was electrically steered to 1000 locations at a pulse repetition frequency of 200 Hz (0.12% duty cycle). Magnetic resonance imaging and histology of the treated tissue revealed a distinct region of necrosis in all samples. Mean lesion volume was 35.6 ± 4.3 mL, generated at 0.9-3.3 mL/min, a speed faster than that of any current ablation method for a large volume. These results suggest that histotripsy has the potential to achieve non-invasive, rapid, homogeneous ablation of a tissue volume., (Copyright © 2017 World Federation for Ultrasound in Medicine and Biology. Published by Elsevier Inc. All rights reserved.)

Histotripsy is a non-invasive, focused ultrasound lesioning technique that can ablate precise volumes of soft tissue using a novel mechanical fractionation mechanism. Previous research suggests that magnetic resonance imaging (MRI) may be a sensitive image-based feedback mechanism for histotripsy. However, there are insufficient data to form some unified understanding of the response of the MR contrast mechanisms in tissues to histotripsy. In this paper, we investigate the response of the MR contrast parameters R1, R2, and the apparent diffusion coefficient (ADC) to various treatment levels of histotripsy in in vitro porcine liver, kidney, muscle, and blood clot as well in formulations of bovine red blood cells suspended in agar gel. We also make a histological analysis of histotripsy lesions in porcine liver. We find that R2 and the ADC are both sensitive to ablation in all materials tested here, and the degree of response varies with tissue type. Correspondingly, under histologic analysis, the porcine liver exhibited various levels of mechanical disruption and necrotic debris that are characteristic of histotripsy. While the area of intact red blood cells and nuclei found within these lesions both decreased with increasing amounts of treatment, the area of red blood cells decreased much more rapidly than the area of intact nuclei. Additionally, the decrease in area of intact red blood cells saturated at the same treatment levels at which the response of the R2 saturated while the area of intact nuclei appeared to vary linearly with the response of the ADC.

Purpose: To demonstrate that MR sequences can detect individual histotripsy bubble clouds formed inside intact tissues., Methods: A line-scan and an EPI sequence were sensitized to histotripsy by inserting a bipolar gradient whose lobes bracketed the lifespan of a histotripsy bubble cloud. Using a 7 Tesla, small-bore scanner, these sequences monitored histotripsy clouds formed in an agar phantom and in vitro porcine liver and brain. The bipolar gradients were adjusted to apply phase with k-space frequencies of 10, 300 or 400 cm -1 . Acoustic pressure amplitude was also varied. Cavitation was simultaneously monitored using a passive cavitation detection system., Results: Each image captured local signal loss specific to an individual bubble cloud. In the agar phantom, this signal loss appeared only when the transducer output exceeded the cavitation threshold pressure. In tissues, bubble clouds were immediately detected when the gradients created phase with k-space frequencies of 300 and 400 cm -1 . When the gradients created phase with a k-space frequency of 10 cm -1 , individual bubble clouds were not detectable until many acoustic pulses had been applied to the tissue., Conclusion: Cavitation-sensitive MR-sequences can detect single histotripsy bubble clouds formed in biologic tissue. Detection is influenced by the sensitizing gradients and treatment history. Magn Reson Med 76:1486-1493, 2016. © 2015 International Society for Magnetic Resonance in Medicine., (© 2015 International Society for Magnetic Resonance in Medicine.)

Background: Raf Kinase Inhibitor Protein1 (RKIP) is a tumor suppressor that is present in several adult tissues. It functions as an inhibitor of both Raf/Mek/Erk and NFĸB signaling when unphosphorylated, but following phosphorylation the ability to inhibit Raf/Mek/Erk signaling is lost and RKIP becomes an activator of G-protein coupled receptor signaling. In neonates and adults, RKIP is known to be expressed in muscle; however, its physiological function is currently unknown., Results: In this study, we show by in situ hybridization and immunofluorescence that RKIP is also expressed in developing chick embryonic muscle, and mouse C2C12 myoblasts. Furthermore, we demonstrate that, in these systems, it functions as an inhibitor of myogenesis: increased levels of RKIP suppress myotube differentiation whereas decreasing RKIP promotes differentiation. Additionally, we show that the ability of RKIP to inhibit myogenesis is dependent upon its phosphorylation state as only the nonphosphorylated form of RKIP suppresses myogenesis., Conclusions: This study, therefore, clearly demonstrates that RKIP has conserved functions as a myogenic inhibitor in both mammalian and avian muscle. Developmental Dynamics 245:902-912, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Mitochondria play a key role in aging, which is a well-established risk factor in amyotrophic lateral sclerosis (ALS). We have previously modeled metabolic dysregulation in ALS using fibroblasts isolated from sporadic ALS (SALS) and familial ALS patients. In the present study, we show that fibroblasts from SALS patients have an altered metabolic response to aging. Control fibroblasts demonstrated increased mitochondrial network complexity and spare respiratory capacity with age which was not seen in the SALS cases. SALS cases displayed an increase in uncoupled mitochondrial respiration, which was not evident in control cases. Unlike SALS cases, controls showed a decrease in glycolysis and an increase in the oxygen consumption rate/extracellular acidification rate ratio, indicating an increased reliance on mitochondrial function. Switching to a more oxidative state by removing glucose with in the culture media resulted in a loss of the mitochondrial interconnectivity and spare respiratory capacity increases observed in controls grown in glucose. Glucose removal also led to an age-independent increase in glycolysis in the SALS cases. This study is, to the best our knowledge, the first to assess the effect of aging on both mitochondrial and glycolytic function simultaneously in intact human fibroblasts and demonstrates that the SALS disease state shifts the cellular metabolic response to aging to a more glycolytic state compared with age-matched control fibroblasts. This work highlights that ALS alters the metabolic equilibrium even in peripheral tissues outside the central nervous system. Elucidating at a molecular level how this occurs and at what stage in the disease process is crucial to understanding why ALS affects cellular energy metabolism and how the disease alters the natural cellular response to aging., (Copyright © 2015 Elsevier Inc. All rights reserved.)