Low-cost prosthetic feet for underserved populations: A comparison of gait analysis and mechanical stiffness.

BACKGROUND: Lower-limb loss is an ongoing cause of disability throughout the world. Despite advancements in prosthetic technologies, there are numerous underserved populations in need of effective low-cost prosthetic foot options. OBJECTIVE: To evaluate the biomechanical performance of several low-cost prosthetic feet, using a combination of instrumented gait analysis and mechanical stiffness testing. STUDY DESIGN: Randomized crossover with additional case study. METHODS: We compared the solid-ankle-cushioned-heel (SACH), Jaipur, and Niagara feet with carbon fiber feet. Mechanical stiffness was evaluated using a cantilever-style bending test at 2 angles that was designed to mimic late stance gait loading. Eight below-knee amputees participated in the gait analysis, which focused on foot and ankle motion and energetics. RESULTS: Metric analysis showed significant differences among feet in ankle motion and power as well as distal-to-shank power, with SACH showing reduced ankle motion and positive work compared with the other feet. Waveform analysis additionally revealed a compensatory knee flexion moment in SACH and a knee extension moment in Niagara and Jaipur during midstance. In mechanical stiffness testing, SACH had the highest stiffness, with Niagara and carbon fiber roughly similar, and Jaipur the most compliant with the greatest hysteresis. CONCLUSIONS: There may be an optimal stiffness range for future prosthesis designs that maximizes propulsive energy. This may be achieved by combining some characteristics of Jaipur and Niagara feet in new designs. Ultimately, optimizing stiffness and energetics for gait biomimicry while maintaining cost, availability, and versatility across cultures will alleviate the effects of limb loss among underserved populations.

Association between sleep quality and physical functioning in adults with Down syndrome: A brief report.

BACKGROUND: Sleep quality is associated with physical functioning in adults, but this has not been examined in those with Down syndrome (DS). High body mass index (BMI) and accelerated aging, both common in adults with DS, may alter the relationship between sleep quality and physical functioning in this population. OBJECTIVE: To examine sleep quality indicators and its association with physical functioning in adults with DS, and whether associations are altered by BMI and age. METHODS: Participants were 15 adults with DS (8 women; age 29 ± 14 years). We evaluated sleep quality over seven days with wrist-worn accelerometers and physical functioning with the timed-up-and-go (TUG) and 6-min walk (6 MW) tests. We examined the associations between sleep quality and physical functioning variables using Spearman's rho. RESULTS: Sleep quality indicators were: total sleep time 407 ± 54 min; latency 26.8 ± 21 min; efficiency 73.9 ± 12 %; wake after sleep onset 122.8 ± 65.2 min; number of awakenings 21.0 ± 6.2; and average length of awakenings 6.1 ± 3 min. Total sleep time and average length of awakenings were significantly associated with 6 MW distances (rho = 0.58 and -0.69; p < 0.05, respectively). After controlling for age and BMI, 6 MW distance was significantly associated with total sleep time, latency, efficiency, and average length of awakenings (rho = 0.56, -0.73, 0.60, and -0.87; p < 0.05, respectively). TUG was significantly associated with total time in bed (rho = 0.71); p < 0.05). CONCLUSIONS: Sleep quality indicators are associated with walking performance in adults with DS. Age and BMI strengthen the relationship between sleep quality and physical functioning.

Best practices and tools for reporting reproducible fluorescence microscopy methods.

Although fluorescence microscopy is ubiquitous in biomedical research, microscopy methods reporting is inconsistent and perhaps undervalued. We emphasize the importance of appropriate microscopy methods reporting and seek to educate researchers about how microscopy metadata impact data interpretation. We provide comprehensive guidelines and resources to enable accurate reporting for the most common fluorescence light microscopy modalities. We aim to improve microscopy reporting, thus improving the quality, rigor and reproducibility of image-based science.

A Fresh Look at the Role of the Coupling of a Discrete State with a Pseudocontinuum State in the Stabilization Method for Characterizing Metastable States.

The stabilization method is widely used to theoretically characterize temporary anions and other systems displaying resonances. In this approach, information about a metastable state is encoded in the interaction of a diabatic discrete state and discretized continuum solutions, the energy of which are varied by scaling the extent of the basis set. In this work, we identify the aspects of the coupling between the discrete state and the discretized continuum states that encode information about the existence of complex stationary points and, hence, complex resonance energies in stabilization graphs. This allows us to design a simple two-level model for extracting complex resonance energies from stabilization graphs. The resulting model is applied to the 2Πg anion state of N2.

Past role and future outlook of the Conservation Reserve Program for supporting honey bees in the Great Plains.

Human dependence on insect pollinators continues to grow even as pollinators face global declines. The Northern Great Plains (NGP), a region often referred to as America's last honey bee (Apis mellifera) refuge, has undergone rapid land-cover change due to cropland expansion and weakened land conservation programs. We conducted a trend analysis and estimated conversion rates of Conservation Reserve Program (CRP) enrollments around bee apiaries from 2006 to 2016 and developed models to identify areas of habitat loss. Our analysis revealed that NGP apiaries lost over 53% of lands enrolled in the CRP, and the rate of loss was highest in areas of high apiary density. We estimated over 163,000 ha of CRP lands in 2006 within 1.6 km of apiaries was converted to row crops by 2012. We also evaluated how alternative scenarios of future CRP acreage caps may affect habitat suitability for supporting honey bee colonies. Our scenario revealed that a further reduction in CRP lands to 7.7 million ha nationally would reduce the number of apiaries in the NGP that meet defined forage criteria by 28% on average. Alternatively, increasing the national cap to 15 million ha would increase the number of NGP apiaries that meet defined forage criteria by 155%. Our scenarios also show that strategic placement of CRP lands near existing apiaries increased the number of apiaries that meet forage criteria by 182%. Our research will be useful for informing the potential consequences of future US farm bill policy and land management in the epicenter of the US beekeeping industry.

7,8-benzoflavone binding to human cytochrome P450 3A4 reveals complex fluorescence quenching, suggesting binding at multiple protein sites.

Human cytochrome P450 (P450) 3A4 is involved in the metabolism of one-half of marketed drugs and shows cooperative interactions with some substrates and other ligands. The interaction between P450 3A4 and the known allosteric effector 7,8-benzoflavone (α-naphthoflavone, αNF) was characterized using steady-state fluorescence spectroscopy. The binding interaction of P450 3A4 and αNF effectively quenched the fluorescence of both the enzyme and ligand. The Hill Equation and Stern-Volmer fluorescence quenching models were used to evaluate binding of ligand to enzyme. P450 3A4 fluorescence was quenched by titration with αNF; at the relatively higher [αNF]/[P450 3A4] ratios in this experiment, two weaker quenching interactions were revealed (Kd 1.8-2.5 and 6.5 µM). A range is given for the stronger interaction since αNF quenching of P450 3A4 fluorescence changed the protein spectral profile: quenching of 315 nm emission was slightly more efficient (Kd 1.8 µM) than the quenching of protein fluorescence at 335 and 355 nm (Kd 2.5 and 2.1 µM, respectively). In the reverse titration, αNF fluorescence was quenched by P450 3A4; at the lower [αNF]/[P450 3A4] ratios here, two strong quenching interactions were revealed (Kd 0.048 and 1.0 µM). Thus, four binding interactions of αNF to P450 3A4 are suggested by this study, one of which may be newly recognized and which could affect studies of drug oxidations by this important enzyme.

The Arp2/3 Complex Is Essential for Distinct Stages of Spine Synapse Maturation, Including Synapse Unsilencing.

UNLABELLED: Dendritic filopodia are actin-rich structures that are thought to contribute to early spine synapse formation; however, the actin regulatory proteins important for early synaptogenesis are poorly defined. Using organotypic hippocampal slice cultures and primary neuron hippocampal cultures from Arp2/3 conditional knock-out mice, we analyze the roles of the Arp2/3 complex, an actin regulator that creates branched actin networks, and demonstrate it is essential for distinct stages of both structural and functional maturation of excitatory spine synapses. Our data show that initially the Arp2/3 complex inhibits the formation of dendritic filopodia but that later during development, the Arp2/3 complex drives the morphological maturation from filopodia to typical spine morphology. Furthermore, we demonstrate that although the Arp2/3 complex is not required for key spine maturation steps, such as presynaptic contact and recruitment of MAGUK (membrane-associated guanylate kinase) scaffolding proteins or NMDA receptors, it is necessary for the recruitment of AMPA receptors. This latter process, also known as synapse unsilencing, is a final and essential step in the neurodevelopment of excitatory postsynaptic synaptogenesis, setting the stage for neuronal interconnectivity. These findings provide the first evidence that the Arp2/3 complex is directly involved in functional maturation of dendritic spines during the developmental period of spinogenesis. SIGNIFICANCE STATEMENT: Excitatory spine synapse formation (spinogenesis) is a poorly understood yet pivotal period of neurodevelopment that occurs within 2-3 weeks after birth. Neurodevelopmental disorders such as intellectual disability and autism are characterized by abnormal spine structure, which may arise from abnormal excitatory synaptogenesis. The initial stage of spinogenesis is thought to begin with the emergence of actin-rich dendritic filopodia that initiate contact with presynaptic axonal boutons. However, it remains enigmatic how actin cytoskeletal regulation directs dendritic filopodial emergence or their subsequent maturation into dendritic spines during development and on into adulthood. In this study, we provide the first evidence that the Arp2/3 complex, a key actin nucleator, is involved in distinct stages of spine formation and is required for synapse unsilencing.

Land-use change reduces habitat suitability for supporting managed honey bee colonies in the Northern Great Plains.

Human reliance on insect pollination services continues to increase even as pollinator populations exhibit global declines. Increased commodity crop prices and federal subsidies for biofuel crops, such as corn and soybeans, have contributed to rapid land-use change in the US Northern Great Plains (NGP), changes that may jeopardize habitat for honey bees in a part of the country that supports >40% of the US colony stock. We investigated changes in biofuel crop production and grassland land covers surrounding ∼18,000 registered commercial apiaries in North and South Dakota from 2006 to 2014. We then developed habitat selection models to identify remotely sensed land-cover and land-use features that influence apiary site selection by Dakota beekeepers. Our study demonstrates a continual increase in biofuel crops, totaling 1.2 Mha, around registered apiary locations in North and South Dakota. Such crops were avoided by commercial beekeepers when selecting apiary sites in this region. Furthermore, our analysis reveals how grasslands that beekeepers target when selecting commercial apiary locations are becoming less common in eastern North and South Dakota, changes that may have lasting impact on pollinator conservation efforts. Our study highlights how land-use change in the NGP is altering the landscape in ways that are seemingly less conducive to beekeeping. Our models can be used to guide future conservation efforts highlighted in the US national pollinator health strategy by identifying areas that support high densities of commercial apiaries and that have exhibited significant land-use changes.

Corrosion protection and improved cytocompatibility of biodegradable polymeric layer-by-layer coatings on AZ31 magnesium alloys.

Composite coatings of electrostatically assembled layer-by-layer anionic and cationic polymers combined with an Mg(OH)2 surface treatment serve to provide a protective coating on AZ31 magnesium alloy substrates. These ceramic conversion coating and layer-by-layer polymeric coating combinations reduced the initial and long-term corrosion progression of the AZ31 alloy. X-ray diffraction and Fourier transform infrared spectroscopy confirmed the successful application of coatings. Potentiostatic polarization tests indicate improved initial corrosion resistance. Hydrogen evolution measurements over a 2 week period and magnesium ion levels over a 1 week period indicate longer range corrosion protection and retention of the Mg(OH)2 passivation layer in comparison to the uncoated substrates. Live/dead staining and DNA quantification were used as measures of biocompatibility and proliferation while actin staining and scanning electron microscopy were used to observe the cellular morphology and integration with the coated substrates. The coatings simultaneously provided improved biocompatibility, cellular adhesion and proliferation in comparison to the uncoated alloy surface utilizing both murine pre-osteoblast MC3T3 cells and human mesenchymal stem cells. The implementation of such coatings on magnesium alloy implants could serve to improve the corrosion resistance and cellular integration of these implants with the native tissue while delivering vital drugs or biological elements to the site of implantation.

Disruption of wave-associated Rac GTPase-activating protein (Wrp) leads to abnormal adult neural progenitor migration associated with hydrocephalus.

Hydrocephalus is the most common developmental disability and leading cause of brain surgery for children. Current treatments are limited to surgical intervention, as the factors that contribute to the initiation of hydrocephalus are poorly understood. Here, we describe the development of obstructive hydrocephalus in mice that are null for Wrp (Srgap3). Wrp is highly expressed in the ventricular stem cell niche, and it is a gene required for cytoskeletal organization and is associated with syndromic and psychiatric disorders in humans. During the postnatal period of progenitor cell expansion and ventricular wall remodeling, loss of Wrp results in the abnormal migration of lineage-tagged cells from the ventricular region into the corpus callosum. Within this region, mutant progenitors appear to give rise to abnormal astroglial cells and induce periventricular lesions and hemorrhage that leads to cerebral aqueductal occlusion. These results indicate that periventricular abnormalities arising from abnormal migration from the ventricular niche can be an initiating cause of noncommunicating hydrocephalus.

WRP/srGAP3 facilitates the initiation of spine development by an inverse F-BAR domain, and its loss impairs long-term memory.

The WAVE-associated Rac GAP, WRP, is thought to regulate key aspects of synapse development and function and may be linked to mental retardation in humans. WRP contains a newly described inverse F-BAR (IF-BAR) domain of unknown function. Our studies show that this domain senses/facilitates outward protrusions analogous to filopodia and that the molecular basis for this is likely explained by a convex lipid-binding surface on the WRP IF-BAR domain. In dendrites the IF-BAR domain of WRP forms a bud on the shaft from which precursors to spines emerge. Loss of WRP in vivo and in vitro results in reduced density of spines. In vivo this is primarily a loss of mushroom-shaped spines. Developmentally, WRP function is critical at the onset of spinogenesis, when dendritic filopodia are prevalent. Finally, because WRP is implicated in mental retardation, behaviors of WRP heterozygous and null mice have been evaluated. Results from these studies confirm that loss of WRP is linked to impaired learning and memory.

Mechanisms of cellular protrusions branch out.

F-BAR domains bind curved membranes and induce membrane invagination. In a recent Cell paper, Guerrier et al. describe an "inverse" F-BAR family member that induces outward curvature and filopodia in migrating neurons. These findings suggest that F-BAR domains are functionally diverse and regulate different types of membrane morphology.