ASU Scholarship Showcase

Sliding clamps are ring-shaped oligomeric proteins that are essential for processive deoxyribonucleic acid replication. Although crystallographic structures of several clamps have been determined, much less is known about clamp structure and dynamics in solution. Here, we characterized the intrinsic solution stability and oligomerization dynamics of the homodimeric Escherichia coli β and the homotrimeric Saccharomyces cerevisiae proliferating cell nuclear antigen (PCNA) clamps using single-molecule approaches. We show that E. coli β is stable in solution as a closed ring at concentrations three orders of magnitude lower than PCNA. The trimeric structure of PCNA results in slow subunit association rates and is largely responsible for the lower solution stability. Despite this large difference, the intrinsic lifetimes of the rings differ by only one order of magnitude. Our results show that the longer lifetime of the E. coli β dimer is due to more prominent electrostatic interactions that stabilize the subunit interfaces.

A new class of highly active solid base catalysts for biodiesel production was developed by creating hierarchically porous aluminosilicate geopolymer with affordable precursors and modifying the material with varying amounts of calcium. For the catalysts containing ≥8 wt% Ca, almost 100% conversion has been achieved in one hour under refluxing conditions with methanol solvent, and the high catalytic activity was preserved for multiple regeneration cycles. Temperature-programed desorption studies of CO2 indicate that the new base catalyst has three different types of base sites on its surface whose strengths are intermediate between MgO and CaO. The detailed powder X-ray diffraction (PXRD) and X-ray photoelectron spectroscopic (XPS) studies show that the calcium ions were incorporated into the aluminosilicate network of the geopolymer structure, resulting in a very strong ionicity of the calcium and thus the strong basicity of the catalysts. Little presence of CaCO3 in the catalysts was indicated from the thermogravimetric analysis (TGA), XPS and Fourier transform infrared spectroscopy (FT-IR) studies, which may contribute to the observed high catalytic activity and regenerability. The results indicate that new geopolymer-based catalysts can be developed for cost-effective biodiesel production.

The purpose of this study was two-fold: 1) demonstrate a technique that can be used to directly estimate the inertial properties of a below-knee prosthesis, and 2) contrast the effects of the proposed technique and that of using intact limb inertial properties on joint kinetic estimates during walking in unilateral, transtibial amputees. An oscillation and reaction board system was validated and shown to be reliable when measuring inertial properties of known geometrical solids. When direct measurements of inertial properties of the prosthesis were used in inverse dynamics modeling of the lower extremity compared with inertial estimates based on an intact shank and foot, joint kinetics at the hip and knee were significantly lower during the swing phase of walking. Differences in joint kinetics during stance, however, were smaller than those observed during swing. Therefore, researchers focusing on the swing phase of walking should consider the impact of prosthesis inertia property estimates on study outcomes. For stance, either one of the two inertial models investigated in our study would likely lead to similar outcomes with an inverse dynamics assessment.

Most current approaches for quantification of RNA species in their natural spatial contexts in single cells are limited by a small number of parallel analyses. Here we report a strategy to dramatically increase the multiplexing capacity for RNA analysis in single cells in situ. In this method, transcripts are detected by fluorescence in situ hybridization (FISH). After imaging and data storage, the fluorescence signal is efficiently removed by photobleaching. This enables the reinitiation of FISH to detect other RNA species in the same cell. Through reiterative cycles of hybridization, imaging and photobleaching, the identities, positions and copy numbers of a large number of varied RNA species can be quantified in individual cells in situ. Using this approach, we analyzed seven different transcripts in single HeLa cells with five reiterative RNA FISH cycles. This approach has the potential to detect over 100 varied RNA species in single cells in situ, which will have wide applications in studies of systems biology, molecular diagnosis and targeted therapies.

Background: The number of days of pedometer or accelerometer data needed to reliably assess physical activity (PA) is important for research that examines the relationship with health. While this important research has been completed in young to middle-aged adults, data is lacking in older adults. Further, data determining the number of days of self-reports PA data is also void. The purpose of this study was to examine the number of days needed to predict habitual PA and sedentary behaviour across pedometer, accelerometer, and physical activity log (PA log) data in older adults.

Methods: Participants (52 older men and women; age = 69.3 ± 7.4 years, range= 55-86 years) wore a Yamax Digiwalker SW-200 pedometer and an ActiGraph 7164 accelerometer while completing a PA log for 21 consecutive days. Mean differences each instrument and intensity between days of the week were examined using separate repeated measures analysis of variance for with pairwise comparisons. Spearman-Brown Prophecy Formulae based on Intraclass Correlations of .80, .85, .90 and .95 were used to predict the number of days of accelerometer or pedometer wear or PA log daily records needed to represent total PA, light PA, moderate-to-vigorous PA, and sedentary behaviour.

Results: Results of this study showed that three days of accelerometer data, four days of pedometer data, or four days of completing PA logs are needed to accurately predict PA levels in older adults. When examining time spent in specific intensities of PA, fewer days of data are needed for accurate prediction of time spent in that activity for ActiGraph but more for the PA log. To accurately predict average daily time spent in sedentary behaviour, five days of ActiGraph data are needed.

Conclusions: The number days of objective (pedometer and ActiGraph) and subjective (PA log) data needed to accurately estimate daily PA in older adults was relatively consistent. Despite no statistical differences between days for total PA by the pedometer and ActiGraph, the magnitude of differences between days suggests that day of the week cannot be completely ignored in the design and analysis of PA studies that involve < 7-day monitoring protocols for these instruments. More days of accelerometer data were needed to determine typical sedentary behaviour than PA level in this population of older adults.

Background: Five accelerometer-derived methods of identifying nonwear and wear time were compared with a self-report criterion in adults ≥ 56 years of age.

Methods: Two hundred participants who reported wearing an Actical™ activity monitor for four to seven consecutive days and provided complete daily log sheet data (i.e., the criterion) were included. Four variables were obtained from log sheets: 1) dates the device was worn; 2) time(s) the participant put the device on each day; 3) time(s) the participant removed the device each day; and 4) duration of self-reported nonwear each day. Estimates of wear and nonwear time using 60, 90, 120, 150 and 180 minutes of consecutive zeroes were compared to estimates derived from log sheets.

Results: Compared with the log sheet, mean daily wear time varied from -84, -43, -24, -14 and -8 min/day for the 60-min, 90-min, 120-min, 150-min and 180-min algorithms, respectively. Daily log sheets indicated 8.5 nonwear bouts per week with 120-min, 150-min and 180-min algorithms estimating 8.2-8.9 nonwear bouts per week. The 60-min and 90-min methods substantially overestimated number of nonwear bouts per week and underestimated time spent in sedentary behavior. Sensitivity (number of compliant days correctly identified as compliant) improved with increasing minutes of consecutive zero counts and stabilized at the 120-min algorithm. The proportion of wear time being sedentary and absolute and proportion of time spent in physical activity of varying intensities were nearly identical for each method.

Conclusions: Utilization of at least 120 minutes of consecutive zero counts will provide dependable population-based estimates of wear and nonwear time, and time spent being sedentary and active in older adults wearing the Actical™ activity monitor.

In spite of well-documented health benefits of vegetarian diets, less is known regarding the effects of these diets on athletic performance. In this cross-sectional study, we compared elite vegetarian and omnivore adult endurance athletes for maximal oxygen uptake (VO2 max) and strength. Twenty-seven vegetarian (VEG) and 43 omnivore (OMN) athletes were evaluated using VO2 max testing on the treadmill, and strength assessment using a dynamometer to determine peak torque for leg extensions. Dietary data were assessed using detailed seven-day food logs. Although total protein intake was lower among vegetarians in comparison to omnivores, protein intake as a function of body mass did not differ by group (1.2 ± 0.3 and 1.4 ± 0.5 g/kg body mass for VEG and OMN respectively, p = 0.220). VO2 max differed for females by diet group (53.0 ± 6.9 and 47.1 ± 8.6 mL/kg/min for VEG and OMN respectively, p < 0.05) but not for males (62.6 ± 15.4 and 55.7 ± 8.4 mL/kg/min respectively). Peak torque did not differ significantly between diet groups. Results from this study indicate that vegetarian endurance athletes’ cardiorespiratory fitness was greater than that for their omnivorous counterparts, but that peak torque did not differ between diet groups. These data suggest that vegetarian diets do not compromise performance outcomes and may facilitate aerobic capacity in athletes.

Multilayer structures of TiO₂/Ag/TiO₂ have been deposited onto flexible substrates by room temperature sputtering to develop indium-free transparent composite electrodes. The effect of Ag thicknesses on optical and electrical properties and the mechanism of conduction have been discussed. The critical thickness (t_c) of Ag mid-layer to form a continuous conducting layer is 9.5 nm and the multilayer has been optimized to obtain a sheet resistance of 5.7 Ω/sq and an average optical transmittance of 90% at 590 nm. The Haacke figure of merit (FOM) for t_c has one of the highest FOMs with 61.4 × 10^-3 Ω^-1/sq.

This manuscript explores the role of embodied views of language comprehension and production in bilingualism and specific language impairment. Reconceptualizing popular models of bilingual language processing, the embodied theory is first extended to this area. Issues such as semantic grounding in a second language and potential differences between early and late acquisition of a second language are discussed. Predictions are made about how this theory informs novel ways of thinking about teaching a second language. Secondly, the comorbidity of speech, language, and motor impairments and how embodiment theory informs the discussion of the etiology of these impairments is examined. A hypothesis is presented suggesting that what is often referred to as specific language impairment may not be so specific due to widespread subclinical motor deficits in this population. Predictions are made about how weaknesses and instabilities in speech motor control, even at a subclinical level, may disrupt the neural network that connects acoustic input, articulatory motor plans, and semantics. Finally, I make predictions about how this information informs clinical practice for professionals such as speech language pathologists and occupational and physical therapists. These new hypotheses are placed within the larger framework of the body of work pertaining to semantic grounding, action-based language acquisition, and action-perception links that underlie language learning and conceptual grounding.

LL-37, a cationic antimicrobial peptide, has numerous immune-modulating effects. However, the identity of a receptor that mediates the responses in immune cells remains uncertain. We have recently demonstrated that LL-37 interacts with the αMI-domain of integrin α_Mβ₂ (Mac-1), a major receptor on the surface of myeloid cells, and induces a migratory response in Mac-1-expressing monocyte/macrophages as well as activation of Mac-1 on neutrophils. Here, we show that LL-37 and its C-terminal derivative supported strong adhesion of various Mac-1-expressing cells, including human embryonic kidney cells stably transfected with Mac-1, human U937 monocytic cells, and murine IC-21 macrophages. The cell adhesion to LL-37 was partially inhibited by specific Mac-1 antagonists, including monoclonal antibody against the α_M integrin subunit and neutrophil inhibitory factor, and completely blocked when anti-Mac-1 antibodies were combined with heparin, suggesting that cell surface heparan sulfate proteoglycans act cooperatively with integrin Mac-1. Coating both gram-negative and gram-positive bacteria with LL-37 significantly potentiated their phagocytosis by macrophages, and this process was blocked by a combination of anti-Mac-1 monoclonal antibody and heparin. Furthermore, phagocytosis by wild-type murine peritoneal macrophages of LL-37-coated latex beads, a model of foreign surfaces, was several fold higher than that of untreated beads. In contrast, LL-37 failed to augment phagocytosis of beads by Mac-1-deficient macrophages. These results identify LL-37 as a novel ligand for integrin Mac-1 and demonstrate that the interaction between Mac-1 on macrophages and bacteria-bound LL-37 promotes phagocytosis.