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The purpose of this study was to examine the relationships between perceived instructor immediacy and student challenge behavior (i.e., procedural, evaluation, power play, practicality) in the college classroom. Participants were 403 students who listened to and reported on a 15 minute guest lecturer in an introductory communication class. Results indicated negative relationships between perceived instructor immediacy and all four types of challenge behavior. Future instructional research should examine instructor behaviors that deter objectionable student behavior. [ABSTRACT FROM AUTHOR]

Tube occlusion is a common and frustrating problem among patients receiving enteral nutrition. These nurses share two methods they found work best for unclogging feeding tubes. [ABSTRACT FROM AUTHOR]

Abstract Background Increasing energy costs and environmental concerns have motivated engineering microbes for the production of "second generation" biofuels that have better properties than ethanol. Results and conclusion Saccharomyces cerevisiae was engineered with an n-butanol biosynthetic pathway, in which isozymes from a number of different organisms (S. cerevisiae, Escherichia coli, Clostridium beijerinckii, and Ralstonia eutropha) were substituted for the Clostridial enzymes and their effect on n-butanol production was compared. By choosing the appropriate isozymes, we were able to improve production of n-butanol ten-fold to 2.5 mg/L. The most productive strains harbored the C. beijerinckii 3-hydroxybutyryl-CoA dehydrogenase, which uses NADH as a co-factor, rather than the R. eutropha isozyme, which uses NADPH, and the acetoacetyl-CoA transferase from S. cerevisiae or E. coli rather than that from R. eutropha. Surprisingly, expression of the genes encoding the butyryl-CoA dehydrogenase from C. beijerinckii (bcd and etfAB) did not improve butanol production significantly as previously reported in E. coli. Using metabolite analysis, we were able to determine which steps in the n-butanol biosynthetic pathway were the most problematic and ripe for future improvement.

Genetic variants in TRIO are associated with neurodevelopmental disorders (NDDs) including schizophrenia (SCZ), autism spectrum disorder (ASD) and intellectual disability. TRIO uses its two guanine nucleotide exchange factor (GEF) domains to activate GTPases (GEF1: Rac1 and RhoG; GEF2: RhoA) that control neuronal development and connectivity. It remains unclear how discrete TRIO variants differentially impact these neurodevelopmental events. Here, we investigate how heterozygosity for NDD-associated Trio variants - +/ K1431M (ASD), +/ K1918X (SCZ), and +/ M2145T (bipolar disorder, BPD) - impact mouse behavior, brain development, and synapse structure and function. Heterozygosity for different Trio variants impacts motor, social, and cognitive behaviors in distinct ways that align with clinical phenotypes in humans. Trio variants differentially impact head and brain size with corresponding changes in dendritic arbors of motor cortex layer 5 pyramidal neurons (M1 L5 PNs). Although neuronal structure was only modestly altered in the Trio variant heterozygotes, we observe significant changes in synaptic function and plasticity. We also identified distinct changes in glutamate synaptic release in +/ K1431M and +/ M2145T cortico-cortical synapses. The TRIO K1431M GEF1 domain has impaired ability to promote GTP exchange on Rac1, but +/ K1431M mice exhibit increased Rac1 activity, associated with increased levels of the Rac1 GEF Tiam1. Acute Rac1 inhibition with NSC23766 rescued glutamate release deficits in +/ K1431M variant cortex. Our work reveals that discrete NDD-associated Trio variants yield overlapping but distinct phenotypes in mice, demonstrates an essential role for Trio in presynaptic glutamate release, and underscores the importance of studying the impact of variant heterozygosity in vivo., Competing Interests: CONFLICT OF INTEREST Authors declare no conflict of interest.

Haemaphysalis longicornis (Neumann), the Asian longhorned tick, is a species native to East Asia, but invasive to Australia, New Zealand, and most recently, the United States. It has spread rapidly across the eastern United States after being established in New Jersey in 2017. Aiding this rapid expansion is the ability of this tick to reproduce parthenogenically and feed on diverse host species. In cattle, this tick can cause heavy burdens and act as a vector for the pathogenic hemoprotozoan parasite Theileria orientalis , genotype Ikeda, creating economic losses that impact the cattle industry. Here, we report Asian longhorned ticks, collected from cattle, a dog, and pastures and morphologically identified at the Oklahoma Animal Disease Diagnostic Laboratory as H. longicornis before molecular confirmation through PCR amplification of the cox1 gene. Blood samples from infested cattle were collected and assessed molecularly for the presence of T. orientalis , with no pathogenic DNA detected. This report describes the first record of H. longicornis in Oklahoma and the farthest westward detection of this tick in the United States to date.

Competing Interests: Conflicts of interest None disclosed.

Telomere shortening is a prominent hallmark of aging and is emerging as a characteristic feature of Myelodysplastic Syndromes (MDS) and Idiopathic Pulmonary Fibrosis (IPF). Optimal telomerase activity prevents progressive shortening of telomeres that triggers DNA damage responses. However, the upstream regulation of telomerase holoenzyme components remains poorly defined. Here, we identify RIOK2, a master regulator of human blood cell development, as a critical transcription factor for telomere maintenance. Mechanistically, loss of RIOK2 or its DNA-binding/transactivation properties downregulates mRNA expression of both TRiC and dyskerin complex subunits that impairs telomerase activity, thereby causing telomere shortening. We further show that RIOK2 expression is diminished in aged individuals and IPF patients, and it strongly correlates with shortened telomeres in MDS patient-derived bone marrow cells. Importantly, ectopic expression of RIOK2 alleviates telomere shortening in IPF patient-derived primary lung fibroblasts. Hence, increasing RIOK2 levels prevents telomere shortening, thus offering therapeutic strategies for telomere biology disorders., (© 2024. The Author(s).)

Neuronal activity is an energy-intensive process that is largely sustained by instantaneous fuel utilization and ATP synthesis. However, how neurons couple ATP synthesis rate to fuel availability is largely unknown. Here, we demonstrate that the metabolic sensor enzyme O-linked N-acetyl glucosamine (O-GlcNAc) transferase regulates neuronal activity-driven mitochondrial bioenergetics in hippocampal and cortical neurons. We show that neuronal activity upregulates O-GlcNAcylation in mitochondria. Mitochondrial O-GlcNAcylation is promoted by activity-driven glucose consumption, which allows neurons to compensate for high energy expenditure based on fuel availability. To determine the proteins that are responsible for these adjustments, we mapped the mitochondrial O-GlcNAcome of neurons. Finally, we determine that neurons fail to meet activity-driven metabolic demand when O-GlcNAcylation dynamics are prevented. Our findings suggest that O-GlcNAcylation provides a fuel-dependent feedforward control mechanism in neurons to optimize mitochondrial performance based on neuronal activity. This mechanism thereby couples neuronal metabolism to mitochondrial bioenergetics and plays a key role in sustaining energy homeostasis., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

The 20S proteasome is a multimeric protease complex that is essential for proteostasis in the cell. Small molecule proteasome inhibitors are approved drugs for various cancers and are advancing clinically as antiparasitics. Although tools and technologies to study the 20S proteasome have advanced, only one probe is commercially available to image proteasome activity. This probe consists of a fluorescently labeled, peptidyl vinyl sulfone that binds to one or more of the catalytic proteasome subunits. Here, we synthesized two, active site-directed epoxyketone probes, LJL-1 and LJL-2, that were based on the peptidyl backbones of the anticancer drugs, carfilzomib and bortezomib, respectively. Each probe was conjugated, via click chemistry, to a bifunctional group comprising 5-carboxytetramethylrhodamine (TAMRA) and biotin to, respectively, visualize and enrich the 20S proteasome from protein extracts of two eukaryotic pathogens, Leishmania donovani and Trichomonas vaginalis . Depending on species, each probe generated a different subunit-binding profile by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and the biotin tag enabled the enrichment of the bound subunits which were then formally identified by proteomics. Species differences in the order of electrophoretic migration by the β subunits were also noted. Finally, both probes reacted specifically with the 20S subunits in contrast to the commercial vinyl sulfone probe that cross reacted with cysteine proteases. LJL-1 and LJL-2 should find general utility in the identification and characterization of pathogen proteasomes, and serve as reagents to evaluate the specificity and mechanism of binding of new antiparasitic proteasome inhibitors., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

We tested the feasibility and preliminary efficacy of an online diet and physical activity program for women with early-stage breast cancer who had completed surgery, chemotherapy, and radiation therapy (ongoing endocrine therapy allowed). Participants with low fruit and vegetable (F/V) consumption and/or low moderate-to-vigorous physical activity (MVPA) levels were randomized to one of two doses - low (one Zoom group session) or high (12 Zoom group sessions) - of an online lifestyle program with the goal of improving F/V intake and MVPA. All participants received eHealth communications (text messages, study website access), a Fitbit, and a WiFi-enabled scale. Primary objectives evaluated feasibility. Secondary objectives compared the 6-month change in F/V intake and MVPA between the two dose groups. Seventy-four women (mean age = 58.4 years; 87% non-Hispanic White; mean time since diagnosis = 4.6 years) were accrued. Among women in the low dose group, 94% attended the single session; among women in the high dose group, 84% attended at least 8 of the 12 sessions. Retention at 6 months was 93%. High relative to low dose participants consumed 1.5 more servings/day of F/V at 6 months (P = 0.007) but MVPA levels did not differ between groups. We successfully implemented an online lifestyle program for early-stage breast cancer survivors. The high dose intervention demonstrated preliminary efficacy in improving F/V consumption in early-stage breast cancer survivors. Future trials can test the intervention in a larger and more diverse population of breast cancer survivors., (© 2024. The Author(s).)

Lower extremity peripheral artery disease (PAD) is a cardiovascular condition manifesting from narrowed or blocked arteries supplying the legs. Gait is impaired in patients with PAD. Recent evidence suggests that walking with carbon fiber ankle foot orthoses (AFOs) can improve patient mobility and delay claudication time. This study aimed to employ advanced biomechanical gait analysis to evaluate the impact of AFO intervention on gait performance among patients with PAD. Patients with claudication had hip, knee, and ankle joint kinetics and kinematics assessed using a cross-over intervention design. Participants walked over the force platforms with and without AFOs while kinematic data was recorded with motion analysis cameras. Kinetics and kinematics were combined to quantify torques and powers during the stance period of the gait cycle. The AFOs effectively reduced the excessive ankle plantar flexion and knee extension angles, bringing the patients' joint motions closer to those observed in healthy individuals. After 3 months of the AFO intervention, the hip range of motion decreased, likely due to changes occurring within the ankle chain. With the assistance of the AFOs, the biological power generation required from the ankle and hip during the push-off phase of walking decreased. Wearing AFOs resulted in increased knee flexor torque during the loading response phase of the gait. Based on this study, AFOs may allow patients with PAD to maintain or improve gait performance. More investigation is needed to fully understand and improve the potential benefits of ankle assistive devices., Competing Interests: Declaration of competing interest Some of the the ankle foot orthosis used by subjects in the study were donated by Ottobock., (Published by Elsevier B.V.)

After spinal cord injury (SCI), re-establishing cellular homeostasis is critical to optimize functional recovery. Central to that response is PERK signaling, which ultimately initiates a pro-apoptotic response if cellular homeostasis cannot be restored. Oligodendrocyte (OL) loss and white matter damage drive functional consequences and determine recovery potential after thoracic contusive SCI. We examined acute (<48 h post-SCI) and chronic (6 weeks post-SCI) effects of conditionally deleting Perk from OLs prior to SCI. While Perk transcript is expressed in many types of cells in the adult spinal cord, its levels are disproportionately high in OL lineage cells. Deletion of OL-Perk prior to SCI resulted in: (1) enhanced acute phosphorylation of eIF2α, a major PERK substrate and the critical mediator of the integrated stress response (ISR), (2) enhanced acute expression of the downstream ISR genes Atf4, Ddit3/Chop, and Tnfrsf10b/Dr5, (3) reduced acute OL lineage-specific Olig2 mRNA, but not neuronal or astrocytic mRNAs, (4) chronically decreased OL content in the spared white matter at the injury epicenter, (5) impaired hindlimb locomotor recovery, and (6) reduced chronic epicenter white matter sparing. Cultured primary OL precursor cells with reduced PERK expression and activated ER stress response showed: (1) unaffected phosphorylation of eIF2α, (2) enhanced ISR gene induction, and (3) increased cytotoxicity. Therefore, OL-Perk deficiency exacerbates ISR signaling and potentiates white matter damage after SCI. The latter effect is likely mediated by increased loss of Perk -/- OLs., (© 2024 Wiley Periodicals LLC.)

Signaling pathways that drive gene expression are typically depicted as having a dozen or so landmark phosphorylation and transcriptional events. In reality, thousands of dynamic post-translational modifications (PTMs) orchestrate nearly every cellular function, and we lack technologies to find causal links between these vast biochemical pathways and genetic circuits at scale. Here we describe the high-throughput, functional assessment of phosphorylation sites through the development of PTM-centric base editing coupled to phenotypic screens, directed by temporally resolved phosphoproteomics. Using T cell activation as a model, we observe hundreds of unstudied phosphorylation sites that modulate NFAT transcriptional activity. We identify the phosphorylation-mediated nuclear localization of PHLPP1, which promotes NFAT but inhibits NFκB activity. We also find that specific phosphosite mutants can alter gene expression in subtle yet distinct patterns, demonstrating the potential for fine-tuning transcriptional responses. Overall, base editor screening of PTM sites provides a powerful platform to dissect PTM function within signaling pathways., (© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Background: Exercise has been shown to improve physical function and quality of life for individuals with cancer. However, low rates of exercise adoption and commonly reported barriers to accessing exercise programming have demonstrated a need for virtual exercise programming in lieu of traditional in-person formats., Objective: The aim of this study was to summarize the existing research on supervised exercise interventions delivered virtually for individuals living with and beyond cancer., Methods: We conducted a scoping review of randomized controlled trials, pilot studies, or feasibility studies investigating virtually supervised exercise interventions for adults either during or after treatment of cancer. The search included EMBASE, MEDLINE, CINAHL, SPORTDiscus, Cochrane Library, and conference abstracts., Results: Fifteen studies were included. The interventions were delivered mostly over Zoom in a group format, with various combinations of aerobic and resistance exercises. Attendance ranged from 78% to 100%, attrition ranged from 0% to 29%, and satisfaction ranged from 94% to 100%. No major adverse events were reported, and only 3 studies reported minor adverse events. Significant improvements were seen in upper and lower body strength, endurance, pain, fatigue, and emotional well-being., Conclusion: Supervised exercise interventions delivered virtually are feasible and may improve physical function for individuals with cancer. The supervision included in these virtual programs promoted similar safety as seen with in-person programming. More randomized controlled trials with large cohorts are needed to validate these findings., Implications for Practice: Individuals living with and beyond cancer can be encouraged to join virtually supervised exercise programs because they are safe, well enjoyed, and may improve physical function and quality of life., Competing Interests: The authors have no funding or conflicts of interest to disclose., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Background: The most common symptom of peripheral artery disease (PAD) is intermittent claudication that involves the calf, thigh, and/or buttock muscles. How the specific location of this leg pain is related to altered gait, however, is unknown., Objectives: We hypothesized that because the location of claudication symptoms uniquely affects different leg muscle groups in people with PAD, this would produce distinctive walking patterns., Methods: A total of 105 participants with PAD and 35 age-matched older volunteers without PAD (CTRL) were recruited. Participants completed walking impairment questionnaires (WIQ), Gardner-Skinner progressive treadmill tests, the six-minute walk test, and we performed an advanced evaluation of the biomechanics of their overground walking. Participants with PAD were categorized into 4 groups according to their stated pain location(s): calf only (C, n = 43); thigh and calf (TC, n = 18); buttock and calf (BC, n = 15); or buttock, thigh, and calf (BTC, n = 29). Outcomes were compared between CTRL, C, TC, BC and BTC groups using a one-way ANOVA with post-hoc comparisons to identify and assess statistically significant differences., Results: There were no significant differences between CTRL, C, TC, BC and BTC groups in distances walked or walking speed when either pain-free or experiencing claudication pain. Each participant with PAD had significantly dysfunctional biomechanical gait parameters, even when pain-free, when compared to CTRL (pain-free) walking data. During pain-free walking, out of the 18 gait parameters evaluated, we only identified significant differences in hip power generation during push-off (in C and TC groups) and in knee power absorption during weight acceptance (in TC and BC groups). There were no between-group differences in gait parameters while people with PAD were walking with claudication pain., Conclusions: Our data demonstrate that PAD affects the ischemic lower extremities in a diffuse manner irrespective of the location of claudication symptoms., Database Registration: ClinicalTrials.gov NCT01970332., (Copyright © 2023 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.)

This study focused on designing and evaluating a bilateral semi-rigid hip exoskeleton. The exoskeleton assisted the hip joint, capitalizing on its proximity to the body's center of mass. Unlike its rigid counterparts, the semi-rigid design permitted greater freedom of movement. A temporal force-tracking controller allowed us to prescribe torque profiles during walking. We ensured high accuracy by tuning control parameters and series elasticity. The evaluation involved experiments with ten participants across ten force profile conditions with different end-timings and peak magnitudes. Our findings revealed a trend of greater reductions in metabolic cost with assistance provided at later timings in stride and at greater magnitudes. Compared to walking with the exoskeleton powered off, the largest reduction in metabolic cost was 9.1%. This was achieved when providing assistance using an end-timing at 44.6% of the stride cycle and a peak magnitude of 0.11 Nm kg -1 . None of the tested conditions reduced the metabolic cost compared to walking without the exoskeleton, highlighting the necessity for further enhancements, such as a lighter and more form-fitting design. The optimal end-timing aligns with findings from other soft hip exosuit devices, indicating a comparable interaction with this prototype to that observed in entirely soft exosuit prototypes.

The composite fermion theory opened a new chapter in understanding many-body correlations through the formation of emergent particles. The formation of two-flux and four-flux composite fermions is well established. While there are limited data linked to the formation of six-flux composite fermions, topological protection associated with them is conspicuously lacking. Here we report evidence for the formation of a quantized and gapped fractional quantum Hall state at the filling factor ν = 9/11, which we associate with the formation of six-flux composite fermions. Our result provides evidence for the most intricate composite fermion with six fluxes and expands the already diverse family of highly correlated topological phases with a new member that cannot be characterized by correlations present in other known members. Our observations pave the way towards the study of higher order correlations in the fractional quantum Hall regime., (© 2024. The Author(s).)

Alterations in the gut-microbiome-brain axis are increasingly being recognized to be involved in Alzheimer's disease (AD) pathogenesis. However, the functional consequences of enteric dysbiosis linking gut microbiota and brain pathology in AD progression remain largely undetermined. The present work investigated the causal role of age-associated temporal decline in butyrate-producing bacteria and butyrate in the etiopathogenesis of AD. Longitudinal metagenomics, neuropathological, and memory analyses were performed in the 3×Tg-AD mouse model. Metataxonomic analyses showed a significant temporal decline in the alpha diversity marked by a decrease in butyrate-producing bacterial communities and a concurrent reduction in cecal butyrate production. Inferred metagenomics analysis identified the bacterial acetyl-CoA pathway as the main butyrate synthesis pathway impacted. Concomitantly, there was an age-associated decline in the transcriptionally permissive acetylation of histone 3 at lysines 9 and 14 (H3K9/K14-Ac) in hippocampal neurons. Importantly, these microbiome-gut-brain changes preceded AD-related neuropathology, including oxidative stress, tau hyperphosphorylation, memory deficits, and neuromuscular dysfunction, which manifest by 17-18 months. Initiation of oral administration of tributyrin, a butyrate prodrug, at 6 months of age mitigated the age-related decline in butyrate-producing bacteria, protected the H3K9/K14-Ac status, and attenuated the development of neuropathological and cognitive changes associated with AD pathogenesis. These data causally implicate age-associated decline in butyrate-producing bacteria as a key pathogenic feature of the microbiome-gut-brain axis affecting the onset and progression of AD. Importantly, the regulation of butyrate-producing bacteria and consequent butyrate synthesis could be a significant therapeutic strategy in the prevention and treatment of AD.

Understanding how small molecules bind to specific protein complexes in living cells is critical to understanding their mechanism-of-action. Unbiased chemical biology strategies for direct readout of protein interactome remodelling by small molecules would provide advantages over target-focused approaches, including the ability to detect previously unknown ligand targets and complexes. However, there are few current methods for unbiased profiling of small molecule interactomes. To address this, we envisioned a technology that would combine the sensitivity and live-cell compatibility of proximity labelling coupled to mass spectrometry, with the specificity and unbiased nature of chemoproteomics. In this manuscript, we describe the BioTAC system, a small-molecule guided proximity labelling platform that can rapidly identify both direct and complexed small molecule binding proteins. We benchmark the system against µMap, photoaffinity labelling, affinity purification coupled to mass spectrometry and proximity labelling coupled to mass spectrometry datasets. We also apply the BioTAC system to provide interactome maps of Trametinib and analogues. The BioTAC system overcomes a limitation of current approaches and supports identification of both inhibitor bound and molecular glue bound complexes., (© 2023. The Author(s).)

Ligation of retinoic acid receptor alpha (RARα) by RA promotes varied transcriptional programs associated with immune activation and tolerance, but genetic deletion approaches suggest the impact of RARα on TCR signaling. Here, we examined whether RARα would exert roles beyond transcriptional regulation. Specific deletion of the nuclear isoform of RARα revealed an RARα isoform in the cytoplasm of T cells. Extranuclear RARα was rapidly phosphorylated upon TCR stimulation and recruited to the TCR signalosome. RA interfered with extranuclear RARα signaling, causing suboptimal TCR activation while enhancing FOXP3 + regulatory T cell conversion. TCR activation induced the expression of CRABP2, which translocates RA to the nucleus. Deletion of Crabp2 led to increased RA in the cytoplasm and interfered with signalosome-RARα, resulting in impaired anti-pathogen immunity and suppressed autoimmune disease. Our findings underscore the significance of subcellular RA/RARα signaling in T cells and identify extranuclear RARα as a component of the TCR signalosome and a determinant of immune responses., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Deep-sea hydrothermal vents offer unique habitats for heat tolerant enzymes with potential new enzymatic properties. Here, we present the novel C11 protease globupain , which was prospected from a metagenome-assembled genome of uncultivated Archaeoglobales sampled from the Soria Moria hydrothermal vent system located on the Arctic Mid-Ocean Ridge. Sequence comparisons against the MEROPS-MPRO database showed that globupain has the highest sequence identity to C11-like proteases present in human gut and intestinal bacteria. Successful recombinant expression in Escherichia coli of the wild-type zymogen and 13 mutant substitution variants allowed assessment of residues involved in maturation and activity of the enzyme. For activation, globupain required the addition of DTT and Ca 2+ . When activated, the 52kDa proenzyme was processed at K 137 and K 144 into a 12kDa light- and 32kDa heavy chain heterodimer. A structurally conserved H 132 /C 185 catalytic dyad was responsible for the proteolytic activity, and the enzyme demonstrated the ability to activate in-trans . Globupain exhibited caseinolytic activity and showed a strong preference for arginine in the P1 position, with Boc-QAR-aminomethylcoumarin (AMC) as the best substrate out of a total of 17 fluorogenic AMC substrates tested. Globupain was thermostable (T m activated enzyme  = 94.51°C ± 0.09°C) with optimal activity at 75°C and pH 7.1. Characterization of globupain has expanded our knowledge of the catalytic properties and activation mechanisms of temperature tolerant marine C11 proteases. The unique combination of features such as elevated thermostability, activity at relatively low pH values, and ability to operate under high reducing conditions makes globupain a potential intriguing candidate for use in diverse industrial and biotechnology sectors., 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 Røyseth, Hurysz, Kaczorowska, Dorawa, Fedøy, Arsın, Serafim, Myers, Werbowy, Kaczorowski, Stokke, O’Donoghue and Steen.)

Response to challenging situations is important to avoid falls, especially after medial perturbations, which require active control. There is a lack of evidence on the relationship between the trunk's motion in response to perturbations and gait stability. Eighteen healthy adults walked on a treadmill at three speeds while receiving perturbations of three magnitudes. Medial perturbations were applied by translating the walking platform to the right at left heel contact. Trunk velocity changes in response to the perturbation were calculated and divided into the initial and the recovery phases. Gait stability after a perturbation was assessed using the margin of stability (MOS) at the first heel contact, MOS mean, and standard deviation for the first five strides after the perturbation onset. Faster speed and smaller perturbations led to a lower deviation of trunk velocity from the steady state, which can be interpreted as an improvement in response to the perturbation. Recovery was quicker after small perturbations. The MOS mean was associated with the trunk's motion in response to perturbations during the initial phase. Increasing walking speed may increase resistance to perturbations, while increasing the magnitude of perturbation leads to greater trunk motions. MOS is a useful marker of resistance to perturbations.

The most common symptom of peripheral artery disease (PAD) is intermittent claudication, which consists of debilitating leg pain during walking. In clinical settings, the presence of PAD is often noninvasively evaluated using the ankle-brachial index and imaging of the arterial supply. Furthermore, various questionnaires and functional tests are commonly used to measure the severity and negative effect of PAD on quality of life. However, these evaluations only provide information on vascular insufficiency and severity of the disease, but not regarding the complex mechanisms underlying walking impairments in patients with PAD. Biomechanical analyses using motion capture and ground reaction force measurements can provide insight into the underlying mechanisms to walking impairments in PAD. This review analyzes the application of biomechanics tools to identify gait impairments and their clinical implications on rehabilitation of patients with PAD. A total of 18 published journal articles focused on gait biomechanics in patients with PAD were studied. This narriative review shows that the gait of patients with PAD is impaired from the first steps that a patient takes and deteriorates further after the onset of claudication leg pain. These results point toward impaired muscle function across the ankle, knee, and hip joints during walking. Gait analysis helps understand the mechanisms operating in PAD and could also facilitate earlier diagnosis, better treatment, and slower progression of PAD.

Background: Peripheral artery disease (PAD) is a cardiovascular disease that limits patients' walking ability. Persistent ankle-foot orthosis (AFO) use may increase the distance patients can walk as well as physical activity., Purpose: The purpose of the study was to determine the implementation and patients' perspectives related to the use or disuse of the AFO intervention six months post-intervention. This study was guided by a semi-structured interview and survey based on the integrated Promoting Action on Research Implementation in Health Services (i-PARIHS) constructs., Design: A convergent mixed methods design was used to evaluate participants' perceptions six months following a three-month AFO intervention. A survey and semi-structured questionnaire based on the i-PARIHS constructs were administered and analyzed., Setting: Vascular surgery clinic and biomechanics research laboratory., Participants: Patients (N = 7; male, 100%; age, 71.9 ± 0.6.7y; body mass index, 29.0 ± 0.5.5; ankle brachial index 0.50 ± 0.17) with claudication completed the study., Interventions: A certified orthotist fit participants with an AFO that was worn for 3 months., Main Outcome Measures: Qualitative analysis of semi-structured interviews and quantitative analysis of the survey., Results: The highest positive ratings were seen in the dimensions of usability and cost-effectiveness. The patients found the AFO device and instructions to wear, easy when starting the intervention and there were no out-of-pocket costs. The lower scores and challenges faced with observability and relative advantage domains indicated issues related to motivation for sustained use of the AFO., Conclusions: Barriers associated with AFO function that prevent common activities and poor health seem to be the biggest issue for not wanting to wear the AFO after the 3-month intervention. Addressing patients' perceptions and challenges to wearing the AFO is essential to increasing compliance and physical activity. Future research should concentrate on understanding the compatibility of orthotic device interventions with the subject's lifestyle., Clinical Trial Registration No: NCT02902211., Competing Interests: Declaration of Competing Interest The authors would like to declare that some ankle-foot orthoses used in this study were donated to the University of Nebraska at Omaha by Ottobock. There are no other competing or financial interests with regard to this work., (Published by Elsevier B.V.)

Cancer is driven by somatic mutations that provide a fitness advantage. While targeted therapies often focus on the mutated gene or its direct downstream effectors, imbalances brought on by cell-state alterations may also confer unique vulnerabilities. In myeloproliferative neoplasms (MPN), somatic mutations in the calreticulin (CALR) gene are disease-initiating through aberrant binding of mutant CALR to the thrombopoietin receptor MPL and ligand-independent activation of JAK-STAT signaling. Despite these mechanistic insights into the pathogenesis of CALR-mutant MPN, there are currently no mutant CALR-selective therapies available. Here, we identified differential upregulation of unfolded proteins, the proteasome and the ER stress response in CALR-mutant hematopoietic stem cells (HSCs) and megakaryocyte progenitors. We further found that combined pharmacological inhibition of the proteasome and IRE1-XBP1 axis of the ER stress response preferentially targets Calr-mutated HSCs and megakaryocytic-lineage cells over wild-type cells in vivo, resulting in an amelioration of the MPN phenotype. In serial transplantation assays following combined proteasome/IRE1 inhibition for six weeks, we did not find preferential depletion of Calr-mutant long-term HSCs. Together, these findings leverage altered proteostasis in Calr-mutant MPN to identify combinatorial dependencies that may be targeted for therapeutic benefit and suggest that eradicating disease-propagating Calr-mutant LT-HSCs may require more sustained treatment., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Background: The design of a randomized pilot trial evaluating the feasibility of two doses of a digital health intervention promoting changes in nutrition and physical activity in breast cancer (BC) survivors is described., Methods: Eligible women were adults with history of early-stage breast cancer and > 60 days post-treatment, consumed <5 servings/day of fruits/vegetables and/or engaged in <150 min/week of aerobic moderate-to-vigorous physical activity, and had internet access. Participants were randomized to 6 months of either a "low" (1 session) or "high" (12 sessions) dose digital health intervention. Zoom-delivered sessions focused on improving diet and physical activity through didactic and experiential classes delivered by a registered dietitian, chef, exercise physiologist, and culinary educator. All study participants received weekly motivational texts, a Fitbit, and study website access. Diet, accelerometry, anthropometric, psychosocial, and biospecimen data were collected remotely at baseline and six months. Primary outcome was feasibility measured via accrual rate, adherence, retention, and acceptability., Results: Recruitment began in December 2019, was suspended in March 2020 due to the COVID-19 pandemic, resumed September 2020, and concluded in January 2021. Women were identified from the local BC registry and flyers posted in the oncology clinic. Of 929 women recruited, 321 completed the screening assessment, and of these, 138 were eligible. A total of 74 women were enrolled and randomized to the study., Conclusion: BC survivors were successfully enrolled in a digital health nutrition and physical activity intervention. If feasible, this intervention will be tested in larger and more diverse populations of cancer survivors., Trial Registration: ClinicalTrials.govNCT04200482., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Supervised exercise therapy (SET) is a conservative non-operative treatment strategy for improving walking performance in patients with peripheral artery disease (PAD). Gait variability is altered in patients with PAD, but the effect of SET on gait variability is unknown. Forty-three claudicating patients with PAD underwent gait analysis before and immediately after a 6-month SET program. Nonlinear gait variability was assessed using sample entropy, and the largest Lyapunov exponent of the ankle, knee, and hip joint angle time series. Linear mean and variability of the range of motion time series for these three joint angles were also calculated. Two-factor repeated measure analysis of variance determined the effect of the intervention and joint location on linear and nonlinear dependent variables. After SET, walking regularity decreased, while the stability remained unaffected. Ankle nonlinear variability had increased values compared with the knee and hip joints. Linear measures did not change following SET, except for knee angle, in which the magnitude of variations increased after the intervention. A six-month SET program produced changes in gait variability toward the direction of healthy controls, which indicates that in general, SET improved walking performance in individuals with PAD.

Peripheral artery disease (PAD) manifests from atherosclerosis, which limits blood flow to the legs and causes changes in muscle structure and function, and in gait performance. PAD is underdiagnosed, which delays treatment and worsens clinical outcomes. To overcome this challenge, the purpose of this study is to develop machine learning (ML) models that distinguish individuals with and without PAD. This is the first step to using ML to identify those with PAD risk early. We built ML models based on previously acquired overground walking biomechanics data from patients with PAD and healthy controls. Gait signatures were characterized using ankle, knee, and hip joint angles, torques, and powers, as well as ground reaction forces (GRF). ML was able to classify those with and without PAD using Neural Networks or Random Forest algorithms with 89% accuracy (0.64 Matthew's Correlation Coefficient) using all laboratory-based gait variables. Moreover, models using only GRF variables provided up to 87% accuracy (0.64 Matthew's Correlation Coefficient). These results indicate that ML models can classify those with and without PAD using gait signatures with acceptable performance. Results also show that an ML gait signature model that uses GRF features delivers the most informative data for PAD classification.

Different levels of arterial occlusive disease (aortoiliac, femoropopliteal, multi-level disease) can produce claudication symptoms in different leg muscle groups (buttocks, thighs, calves) in patients with peripheral artery disease (PAD). We tested the hypothesis that different locations of occlusive disease uniquely affect the muscles of PAD legs and produce distinctive patterns in the way claudicating patients walk. Ninety-seven PAD patients and 35 healthy controls were recruited. PAD patients were categorized to aortoiliac, femoropopliteal and multi-level disease groups using computerized tomographic angiography. Subjects performed walking trials both pain-free and during claudication pain and joint kinematics, kinetics, and spatiotemporal parameters were calculated to evaluate the net contribution of the calf, thigh and buttock muscles. PAD patients with occlusive disease affecting different segments of the arterial tree (aortoiliac, femoropopliteal, multi-level disease) presented with symptoms affecting different muscle groups of the lower extremity (calves, thighs and buttocks alone or in combination). However, no significant biomechanical differences were found between PAD groups during the pain-free conditions with minimal differences between PAD groups in the claudicating state. All statistical differences in the pain-free condition occurred between healthy controls and one or more PAD groups. A discriminant analysis function was able to adequately predict if a subject was a control with over 70% accuracy, but the function was unable to differentiate between PAD groups. In-depth gait analyses of claudicating PAD patients indicate that different locations of arterial disease produce claudication symptoms that affect different muscle groups across the lower extremity but impact the function of the leg muscles in a diffuse manner generating similar walking impairments., Competing Interests: The authors have declared that no competing interests exist.

Master regulators, such as the hematopoietic transcription factor (TF) GATA1, play an essential role in orchestrating lineage commitment and differentiation. However, the precise mechanisms by which such TFs regulate transcription through interactions with specific cis-regulatory elements remain incompletely understood. Here, we describe a form of congenital hemolytic anemia caused by missense mutations in an intrinsically disordered region of GATA1, with a poorly understood role in transcriptional regulation. Through integrative functional approaches, we demonstrate that these mutations perturb GATA1 transcriptional activity by partially impairing nuclear localization and selectively altering precise chromatin occupancy by GATA1. These alterations in chromatin occupancy and concordant chromatin accessibility changes alter faithful gene expression, with failure to both effectively silence and activate select genes necessary for effective terminal red cell production. We demonstrate how disease-causing mutations can reveal regulatory mechanisms that enable the faithful genomic targeting of master TFs during cellular differentiation., (© 2022 by The American Society of Hematology.)

Retrons are bacterial retroelements that produce single-stranded, reverse-transcribed DNA (RT-DNA) that is a critical part of a newly discovered phage defense system. Short retron RT-DNAs are produced from larger, structured RNAs via a unique 2'-5' initiation and a mechanism for precise termination that is not yet understood. Interestingly, retron reverse transcriptases (RTs) typically lack an RNase H domain and, therefore, depend on endogenous RNase H1 to remove RNA templates from RT-DNA. We find evidence for an expanded role of RNase H1 in the mechanism of RT-DNA termination, beyond the mere removal of RNA from RT-DNA:RNA hybrids. We show that endogenous RNase H1 determines the termination point of the retron RT-DNA, with differing effects across retron subtypes, and that these effects can be recapitulated using a reduced, in vitro system. We exclude mechanisms of termination that rely on steric effects of RNase H1 or RNA secondary structure and, instead, propose a model in which the tertiary structure of the single-stranded RT-DNA and remaining RNA template results in termination. Finally, we show that this mechanism affects cellular function, as retron-based phage defense is weaker in the absence of RNase H1., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Patients with peripheral artery disease (PAD) have significantly reduced lower extremity muscle strength compared with healthy individuals as measured during isolated, single plane joint motion by isometric and isokinetic strength dynamometers. Alterations to the force contribution of muscles during walking caused by PAD are not well understood. Therefore, this study used simulations with PAD biomechanics data to understand lower extremity muscle functions in patients with PAD during walking and to compare that with healthy older individuals. A total of 12 patients with PAD and 10 age-matched healthy older controls walked across a 10-meter pathway with reflective markers on their lower limbs. Marker coordinates and ground reaction forces were recorded and exported to OpenSim software to perform gait simulations. Walking velocity, joint angles, muscle force, muscle power, and metabolic rate were calculated and compared between patients with PAD and healthy older controls. Our results suggest that patients with PAD walked slower with less hip extension during propulsion. Significant force and power reductions were observed in knee extensors during weight acceptance and in plantar flexors and hip flexors during propulsion in patients with PAD. The estimated metabolic rate of walking during stance was not different between patients with PAD and controls. This study is the first to analyze lower limb muscular responses during walking in patients with PAD using the OpenSim simulation software. The simulation results of this study identified important information about alterations to muscle force and power during walking in those with PAD., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The metabolic rate of walking can be reduced by applying a constant forward force at the center of mass. It has been shown that the metabolically optimal constant force magnitude minimizes propulsion ground reaction force at the expense of increased braking. This led to the hypothesis that selectively assisting propulsion could lead to greater benefits. We used a robotic waist tether to evaluate the effects of forward forces with different timings and magnitudes. Here, we show that it is possible to reduce the metabolic rate of healthy participants by 48% with a greater efficiency ratio of metabolic cost reduction per unit of net aiding work compared with other assistive robots. This result was obtained using a sinusoidal force profile with peak timing during the middle of the double support. The same timing could also reduce the metabolic rate in patients with peripheral artery disease. A model explains that the optimal force profile accelerates the center of mass into the inverted pendulum movement during single support. Contrary to the hypothesis, the optimal force timing did not entirely coincide with propulsion. Within the field of wearable robotics, there is a trend to use devices to mimic biological torque or force profiles. Such bioinspired actuation can have relevant benefits; however, our results demonstrate that this is not necessarily optimal for reducing metabolic rate.

Cells integrate complex cytokine cues and other inflammatory stimuli through activation of the signal transducers and activators of transcription (STAT) family of transcription factors to drive the appropriate anti-microbial, inflammatory, and resolving functions. Here, we discuss recent progress in our understanding of mechanisms supporting STAT functional diversity. Signaling component availability and the strength of receptor and STAT interactions emerge as important determinants of immune function. The resultant dynamics of STAT activation, together with stimulus-specific variation in STAT post-translationally modified states, will impact downstream binding partners to support transcription of distinct gene subsets. Understanding how context-dependent STAT function is encoded to dictate cytokine specificity, crosstalk, and control of inflammation will guide therapeutic efforts to selectively perturb STAT-regulated responses., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Objective: The aim of this scoping review was to identify information on compliance with wearing orthoses and other supportive devices, to discuss the barriers to adherence, and to suggest strategies for improvement based on these findings., Methods: Online databases of PubMed, Web of Science, and the Cochrane Library were searched for articles about patients' compliance with regard to lower limb assistive devices. In addition, a methodological quality control process was conducted. Studies were included if in the English language and related to compliance and adherence to the lower limb assistive device. Exclusion was based on first reading the abstract and then the full manuscript confirming content was not related to orthotic devices and compliance., Results: Twelve studies were included. The data revealed between 6% and 80% of patients were not using a prescribed device. Barriers to the use of the orthotic device included medical, functional, device properties and lack of proper fit. Strategies for improved compliance included better communication between patient and clinician, patient education, and improved comfort and device esthetics., Conclusions: Individualized orthotic adjustments, rehabilitation, and patient education were promising for increasing adherence. Despite positive aspects of improvements in gait, balance in elderly, and a sense of security produced by using assistive devices, compliance remains less than ideal due to barriers. As compliance in recent studies has not improved, continued work in this area is essential to realize the benefits of technological advances in orthotic and assistive devices., (Copyright © 2022. Published by Elsevier Inc.)

The roll-over shape (ROS) effectively characterizes the lower limb's ability to roll forward during the single-limb support phase of human walking. ROS is modelled as an optimally fitted circular arc to the center of pressure (CoP) data transformed in the shank/leg-based local coordinate system. The commonly used method for optimal fitting of ROS is complex to implement and eliminates inherent individual variability in the ROS parameters during walking. We propose and validate a novel computerized method for optimal fitting of roll-over shapes of the lower limb during walking. Gait data of a healthy individual from Winter's book was used to generate ankle-foot and knee-ankle-foot roll-over shapes using the proposed method. The goodness of fit and form of both the roll-over shapes were validated with the literature. To test the robustness of the proposed technique, small random perturbations were introduced to the transformed CoP data and the effect of these small changes in the data on the ROS parameters was studied. The ROS parameters such as radius, arc length, subtended arc angle, and horizontal and vertical shift in the arc center did not change substantially with small changes in the CoP data. The proposed method is computationally efficient, and easy to implement for optimal fitting and characterization of ROS., (Copyright © 2022. Published by Elsevier Ltd.)

Anemia is a major comorbidity in aging, chronic kidney and inflammatory diseases, and hematologic malignancies. However, the transcriptomic networks governing hematopoietic differentiation in blood cell development remain incompletely defined. Here we report that the atypical kinase RIOK2 (right open reading frame kinase 2) is a master transcription factor (TF) that not only drives erythroid differentiation, but also simultaneously suppresses megakaryopoiesis and myelopoiesis in primary human stem and progenitor cells. Our study reveals the previously uncharacterized winged helix-turn-helix DNA-binding domain and two transactivation domains of RIOK2 that are critical to regulate key hematopoietic TFs GATA1, GATA2, SPI1, RUNX3 and KLF1. This establishes RIOK2 as an integral component of the transcriptional regulatory network governing human hematopoietic differentiation. Importantly, RIOK2 mRNA expression significantly correlates with these TFs and other hematopoietic genes in myelodysplastic syndromes, acute myeloid leukemia and chronic kidney disease. Further investigation of RIOK2-mediated transcriptional pathways should yield therapeutic approaches to correct defective hematopoiesis in hematologic disorders., (© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Objective: Supervised exercise therapy (SET) is a first-line treatment for patients with peripheral artery disease (PAD). The efficacy of SET is most commonly expressed by significant statistical improvement of parameters that do not clarify how each individual patient will benefit from SET. This study examined the minimal clinically important difference (MCID) in walking speed in claudicating patients with PAD after SET., Methods: A total of 63 patients with PAD-related claudication (Fontaine stage II PAD) participated in a 6-month SET program. Self-selected walking speed was measured before and after SET. Distribution and anchor-based approaches were used to estimate the MCID for small and substantial improvement. The ability to walk one block and the ability to climb one flight of stairs questions were chosen as anchor questions from the Medical Outcomes Study 36-item Short Form questionnaire. Receiver operating characteristics curve analyses were performed to detect the threshold for MCID in walking speed after treatment., Results: The distribution-based method estimated 0.03 m/s as a small improvement and 0.08 m/s as a substantial improvement after SET. Small and substantial improvements according to the anchor question walking one block were 0.05 m/s and 0.15 m/s, respectively. For the climbing one flight of stairs anchor question, 0.10 m/s was a small improvement. Receiver operating characteristics curve analyses identified an increase of 0.04 m/s and 0.03 m/s for improvement based on walking one block and climbing one flight of stairs, respectively., Conclusions: We report our findings for the MCID for walking speed among claudicating patients receiving SET. Claudicating patients who increase walking speed of 0.03 m/s or greater are more likely to experience a meaningful improvement in walking impairment than those who do not. The MCID reported in this study can serve as a benchmark for clinicians to develop goals and interpret clinically meaningful progress in the care of claudicating patients with PAD., (Copyright © 2021 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)