Clinical Potential of Transcranial Focused Ultrasound for Neurorehabilitation in Pediatric Cancer Survivors.

Cancer survivors are at a high risk for treatment-related late effects, particularly neurocognitive impairment in the attention and executive function domains. These can be compounded in pediatric populations still undergoing neural development, which has increased interest in survivorship studies and neurorehabilitation approaches to mitigate these effects. Cognitive training regimens have shown promise as a therapeutic intervention for improving cognitive function. Therapist-guided and computerized training programs with adaptive paradigms have been successfully implemented in pediatric populations, with positive outcomes on attention and working memory. Another interventional approach is neuromodulation to alter plasticity. Transcranial electrical stimulation can modulate cortical surface activity, and cranial nerve stimulation alters autonomic activity in afferent brainstem pathways. However, they are more systemic in nature and have diffuse spatial targeting. Transcranial focused ultrasound (tFUS) modulation overcomes these limitations with high spatial specificity and the ability to target deeper brain regions. In this review, we discuss the efficacy of tFUS for modulating specific brain regions and its potential utility to augment cognitive training programs as a complementary intervention.

Dynamic peripheral nerve stimulation can produce cortical activation similar to punctate mechanical stimuli.

During contact, phasic and tonic responses provide feedback that is used for task performance and perceptual processes. These disparate temporal dynamics are carried in peripheral nerves, and produce overlapping signals in cortex. Using longitudinal intrafascicular electrodes inserted into the median nerve of a nonhuman primate, we delivered composite stimulation consisting of onset and release bursts to capture rapidly adapting responses and sustained stochastic stimulation to capture the ongoing response of slowly adapting receptors. To measure the stimulation's effectiveness in producing natural responses, we monitored the local field potential in somatosensory cortex. We compared the cortical responses to peripheral nerve stimulation and vibrotactile/punctate stimulation of the fingertip, with particular focus on gamma band (30-65 Hz) responses. We found that vibrotactile stimulation produces consistently phase locked gamma throughout the duration of the stimulation. By contrast, punctate stimulation responses were phase locked at the onset and release of stimulation, but activity maintained through the stimulation was not phase locked. Using these responses as guideposts for assessing the response to the peripheral nerve stimulation, we found that constant frequency stimulation produced continual phase locking, whereas composite stimulation produced gamma enhancement throughout the stimulus, phase locked only at the onset and release of the stimulus. We describe this response as an "Appropriate Response in the gamma band" (ARγ), a trend seen in other sensory systems. Our demonstration is the first shown for intracortical somatosensory local field potentials. We argue that this stimulation paradigm produces a more biomimetic response in somatosensory cortex and is more likely to produce naturalistic sensations for readily usable neuroprosthetic feedback.

High regrowth of potato crop wild relative genotypes after cryogenic storage.

Potatoes are consumed by millions of people and are the survival food in several countries. Cultivated varieties of potato (Solanum tubersosum L.) are results of selection and crossing of many wild species. Only 8-13% of wild potato species used for food are preserved by either in situ or ex situ methods. The U.S. National Potato Germplasm Collection maintains over 5900 accessions, of which 75% are crop wild relatives (CWR). The objective of the study was to investigate regrowth of cryogenically stored clonal propagules (shoot tips) of selected CWR accessions maintained in the collection. Sixty-nine accessions from 30 Solanum species and six accessions that are not yet assigned to a species were cryopreserved by a droplet vitrification method at the NLGRP. The post cryopreservation regrowth varied from 40 to 100% (average 68%) but was not significantly different between the tested accessions. Regrowth of six accessions tested after 10 years of cryogenic storage was between 35 and 90% (average 66%) and was significantly different from their initial regrowth (average 87%); the largest viability loss was in S. condolleanum; but for the other five accessions the regrowth was between 45 and 90% (average 72%) and suggested at least 10 years of successful storage in LN was possible. Twelve potato wild species cryopreserved in this study were reported in literature as important for developing cultivated varieties for changed weather conditions.

Adapting wine grape production to climate change through canopy architecture manipulation and irrigation in warm climates.

Grape growing regions are facing constant warming of the growing season temperature as well as limitations on ground water pumping used for irrigating to overcome water deficits. Trellis systems are utilized to optimize grapevine production, physiology, and berry chemistry. This study aimed to compare 6 trellis systems with 3 levels of applied water amounts based on different replacements of crop evapotranspiration (ETc) in two consecutive seasons. The treatments included a vertical shoot position (VSP), two modified VSPs (VSP60 and VSP80), a single high wire (SH), a high quadrilateral (HQ), and a Guyot pruned VSP (GY) combined with 25%, 50%, and 100% ETc water replacement. The SH had greater yields, whereas HQ was slower to reach full production potential. At harvest in both years, the accumulation of anthocyanin derivatives was enhanced in SH, whereas VSPs decreased them. As crown porosity increased (mostly VSPs), berry flavonol concentration and likewise molar % of quercetin in berries increased. Conversely, as leaf area increased, total flavonol concentration and molar % of quercetin decreased, indicating a preferential arrangement of leaf area along the canopy for overexposure of grape berry with VSP types. The irrigation treatments revealed linear trends for components of yield, where greater applied water resulted in larger berry size and likewise greater yield. 25% ETc was able to increase berry anthocyanin and flavonol concentrations. Overall, this study evidenced the efficiency of trellis systems for optimizing production and berry composition in Californian climate, also, the feasibility of using flavonols as the indicator of canopy architecture.

Effect of Tactile Sensory Substitution on the Proprioceptive Error Map of the Arm.

Proprioceptive error of estimated fingertip position in two-dimensional space is reduced with the addition of tactile stimulation to the fingertip. This tactile input does not disrupt the subjects' estimation strategy, as the individual error vector maps maintain their overall geometric structure. This relationship suggests an integration of proprioception and tactile sensory information to enhance proprioceptive estimation. To better understand this multisensory integration, we explored the effect of electrotactile and vibrotactile stimulation to the fingertips in place of actual contact, thus limiting interaction forces. This allowed us to discern any proprioceptive estimation improvement that arose from purely tactile stimulation. Ten right-handed and ten left-handed subjects performed a simple right-handed proprioceptive estimation task under four tactile feedback conditions: hover, touch, electrotactile, and vibrotactile. Target sets were generated for each subject, persisted across all feedback modalities, and targets were presented in randomized orders. Error maps across the workspace were generated using polynomial models of the subjects' responses. Error maps did not change shape between conditions for any right-handed subjects and changed for a single condition for two left-handed subjects. Non-parametric statistical analysis of the error magnitude shows that both modes of sensory substitution significantly reduce error for right-handed subjects, but not to the level of actual touch. Left-handed subjects demonstrated increased error for all feedback conditions compared to hover. Compared to right-handed subjects, left-handed subjects demonstrated more error in each condition except the hover condition. This is consistent with the hypothesis that the non-dominant hand is specialized for position control, while the dominant is specialized for velocity. Notably, our results suggest that non-dominant hand estimation strategies are hindered by stimuli to the fingertip. We conclude that electrotactile and vibrotactile sensory substitution only succeed in multisensory integration when applied to the dominant hand. These feedback modalities do not disrupt established dominate hand proprioceptive error maps, and existing strategies adapt to the novel input and minimize error. Since actual touch provides the best error reduction, sensory substitution lacks some unidentified beneficial information, such as familiarity or natural sensation. This missing component could also be what confounds subjects using their non-dominant hand for positional tasks.

Determining the effect of pretreatments on freeze resistance and survival of cryopreserved temperate fruit tree dormant buds.

Freeze resistance is critical to successful dormant bud (DB) cryopreservation, and is affected by genotype, environmental conditions, dormancy phase and processing techniques. Pretreatment induced freeze resistance may contribute to more successful and efficient protocols for cryopreserving DB. Differential thermal analysis (DTA) was used to quantify the effects of cryopreservation pretreatments on freeze resistance of dormant budwood. Low temperature exotherm (LTE) profiles created by DTA could rapidly identify pretreatments that are contributing to increased freeze resistance in tree fruit species. In this study, DTA was used to help elucidate the effects of varying pretreatments (sucrose, desiccation and their combination) on peach, a model crop in tree fruit physiology that has shown little cryosurvival using the DB method in the past. Post cryopreservation recovery trials using an antimicrobial forced bud development (AFBD) protocol evaluated the ability of selected pretreatments, that improved freeze resistance based on DTA, to improve recovery of dormant budwood of various deciduous tree fruit and nut species. Precryogenic exposure to sucrose solution (5.0 M, 96 h), desiccation to 30% moisture content (MC) and their combination tested for their efficacy on improving postcryogenic viability in peach, apricot, sweet cherry, little walnut, black walnut, English walnut, apple, and pear. Among the different pretreatments tested, desiccation to 30% MC had the greatest impact on increasing freeze resistance and cryosurvival across most fruit species tested and little walnut. Gradual reduction of MC (from 40 to 25%) levels increased freeze resistance in peach (R2=0.95) and increased some recovery outcomes (leaf, shoot and bud swell), however, this was not correlated with equal cryorecovery outcomes as severe bud cracking was observed. Overall, our approach linking freeze resistance and preconditioning treatments could help establish efficient species-specific cryopreservation protocols for a number of important temperate woody crops which could be recovered as complete plants by coupling AFBD and plant tissue culture.

Antimicrobial forcing solution improves recovery of cryopreserved temperate fruit tree dormant buds.

Dormant bud cryogenic preservation is a cost- and labor-efficient method of genetic resource backup compared to in vitro derived meristem shoots cryopreservation. While protocols have been developed for cryopreserving apple dormant buds, effective and reproducible protocols are yet to be developed for several temperate fruit and nut species. Dormant bud cryopreservation typically requires material to be grafted to evaluate viability and recover a plant. Forced bud development has been used on a very limited scale for cryostored dormant budwood recovery, however, it provides a labor-efficient alternative viability assessment. To increase the utility of this approach, regrowth must be optimized to allow complete plant recovery. We hypothesized that bacterial attacks are limiting regrowth, thus, an antimicrobial forcing solution can maximize regrowth potential. This study examined the effects of an antimicrobial forcing solution (8-hydroxyquinoline citrate and sucrose, 8-HQC) on the cryosurvival and recovery of dormant buds of fruit (Malus x domestica, Prunus armeniaca, Prunus avium, Prunus persica, Pyrus communis), and nut species (Juglans regia, Juglans nigra, Juglans microcarpa). Recovery and shoot development were significantly improved for all the fruit and one nut species (J. microcarpa) treated with the 8-HQC, compared to standard recovery under high humidity alone (P < 0.001). Additionally, this post cryo recovery approach led to successful in vitro shoot tip establishment across all surviving fruit species. 8-HQC embedded forced bud development method increased viability and efficiency for existing cryostored material and can be used as a benchmark to develop protocols for different crops that could potentially lead to complete plant recovery.

Anisotropic Psychophysical Trends in the Discrimination of Tactile Direction in a Precision Grip.

Tactile cues arising from interactions with objects have a sense of directionality which affects grasp. Low latency responses to varied grip perturbations indicate that grasp safety margins are exaggerated in certain directions and conditions. In a grip with the ulnar-radial axis vertical, evidence suggests that distal and downward directions are more sensitive to task parameters and have larger safety margins. This suggests that, for the purpose of applying forces with the fingers, reference frames with respect to the hand and gravity are both in operation. In this experiment, we examined human sensitivities to the direction of tactile movement in the context of precision grip in orientations either orthogonal to or parallel to gravity. Subjects performed a two-alternative-forced-choice task involving a textured cube which moved orthogonal to their grip axis. Subjects' arms were placed in a brace that allowed for finger movement but minimized arm movement. Movement of thumb and index joints were monitored via PhaseSpace motion capture. The subject was presented with a textured cube and instructed to lightly grasp the cube, as if it were slipping. In each trial the object was first translated 1 cm in 0° (proximal), 90° (radial), 180° (distal), or 270° (ulnar) and returned to its origin. This primary stimulus was immediately followed by a 10 mm secondary stimulus at a random 5° interval between -30° and 30° of the primary stimulus. Response from the subject after each pair of stimuli indicated whether the test direction felt the same as or different from the primary stimulus. Traditional bias and sensitivity analyses did not provide conclusive results but suggested that performance is best in the ulnar-radial axis regardless of gravity. Modeling of the response curve generated a detection threshold for each primary stimulus. Lower thresholds, indicating improved detection, persisted in the ulnar-radial axis. Anisotropic thresholds of increased detection appear to coincide with digit displacement and appear to be independent of the grasp orientation.

Tunable substrates improve imaging of viruses and cancer proteins.

Recent breakthroughs in cryo-electron microscopy imaging technology provide an unprecedented view of biology at the nanoscale. To complement these technical advances, here we demonstrate the use of tunable substrates to streamline the isolation of biological entities from human cells. We have tested the capacity of tunable microchip devices using a variety of samples including virus assemblies and the breast cancer susceptibility protein (BRCA1) produced in cancer cells. Overall, microchip applications may shed light on ill-defined clinical issues related to molecular disease mechanisms.

Twig pre-harvest temperature significantly influences effective cryopreservation of Vaccinium dormant buds.

Cryopreservation of temperate woody-plant material by dormant buds is less expensive than using shoot tips isolated from tissue cultured plants; however currently, dormant buds are used only for preservation of selected temperate tree and shrub species. Using dormant buds could be an efficient strategy for long-term preservation of blueberry (Vaccinium L.) genetic resources. In this study, viability of V. hybrid 'Northsky' (PI 554943) dormant buds was evaluated at 30 harvest dates over three consecutive fall/winter seasons to determine the optimal harvest time that promotes high post cryopreservation viability. Twigs with dormant buds were cut into 70 mm segments containing at least two nodes, desiccated, slowly cooled, stored in liquid nitrogen vapor and tested for post-cryopreservation regrowth. The highest regrowth of cryopreserved dormant buds was observed for buds harvested in mid-December and during the first half of January. Pearson's correlation coefficients were computed to evaluate the association between bud characteristics and viability at harvest date and logistic regression models were fit to test the ability of twig characteristics and temperatures to predict post cryopreservation bud viability. Post-cryopreservation viability was negatively correlated (p < 0.05) with average minimum, maximum and daily mean temperature preceding the bud harvest but was not correlated with the dormant bud initial and end moisture content, twig diameter, the number of dormant buds/cm of twig length and the number of days in desiccation. Regression tree analysis suggested post-cryopreservation viability to be between 52 and 80% for dormant buds harvested after a 10 day average maximum air temperature of <11.2 °C. Pre-harvest air temperature was a significant indicator of optimal dormant bud harvest time to produce adequate viability for long term preservation of blueberry genetic resources.

A Molecular Toolkit to Visualize Native Protein Assemblies in the Context of Human Disease.

We present a new molecular toolkit to investigate protein assemblies natively formed in the context of human disease. The system employs tunable microchips that can be decorated with switchable adaptor molecules to select for target proteins of interest and analyze them using molecular microscopy. Implementing our new streamlined microchip approach, we could directly visualize BRCA1 gene regulatory complexes from patient-derived cancer cells for the first time.

A robot hand testbed designed for enhancing embodiment and functional neurorehabilitation of body schema in subjects with upper limb impairment or loss.

Many upper limb amputees experience an incessant, post-amputation "phantom limb pain" and report that their missing limbs feel paralyzed in an uncomfortable posture. One hypothesis is that efferent commands no longer generate expected afferent signals, such as proprioceptive feedback from changes in limb configuration, and that the mismatch of motor commands and visual feedback is interpreted as pain. Non-invasive therapeutic techniques for treating phantom limb pain, such as mirror visual feedback (MVF), rely on visualizations of postural changes. Advances in neural interfaces for artificial sensory feedback now make it possible to combine MVF with a high-tech "rubber hand" illusion, in which subjects develop a sense of embodiment with a fake hand when subjected to congruent visual and somatosensory feedback. We discuss clinical benefits that could arise from the confluence of known concepts such as MVF and the rubber hand illusion, and new technologies such as neural interfaces for sensory feedback and highly sensorized robot hand testbeds, such as the "BairClaw" presented here. Our multi-articulating, anthropomorphic robot testbed can be used to study proprioceptive and tactile sensory stimuli during physical finger-object interactions. Conceived for artificial grasp, manipulation, and haptic exploration, the BairClaw could also be used for future studies on the neurorehabilitation of somatosensory disorders due to upper limb impairment or loss. A remote actuation system enables the modular control of tendon-driven hands. The artificial proprioception system enables direct measurement of joint angles and tendon tensions while temperature, vibration, and skin deformation are provided by a multimodal tactile sensor. The provision of multimodal sensory feedback that is spatiotemporally consistent with commanded actions could lead to benefits such as reduced phantom limb pain, and increased prosthesis use due to improved functionality and reduced cognitive burden.

Rotavirus core shell subdomains involved in polymerase encapsidation into virus-like particles.

The triple-layered rotavirus virion encases an 11-segmented, dsRNA genome and 11-12 copies of the viral polymerase (VP1). VP1 transcribes and replicates the genome while tethered beneath the VP2 core shell. Genome replication (i.e. minus-strand RNA synthesis) by VP1 occurs in association with core assembly. During this process, VP2 directly engages VP1, thereby (i) packaging the polymerase into a nascent core and (ii) triggering the enzyme to initiate minus-strand RNA synthesis on bound plus-strand RNA templates. Recent work has shed light on VP2 regions important for VP1 enzymic activity. In the current study, we sought to investigate VP2 subdomains involved in the encapsidation of VP1 into recombinant virus-like particles (VLPs), which are formed of VP2 and the middle layer virion protein (VP6). We showed that strain SA11 VLPs efficiently encapsidated SA11 VP1, but not the genetically divergent Bristol VP1. VLPs made with an SA11 VP2 mutant lacking residues 1-10 of the amino-terminal domain (NTD) were still able to encapsidate VP1; however, removal of the entire NTD (residues 1-102) completely abolished polymerase packaging. We also showed that a chimeric VP2 protein containing the NTD and dimer-forming subdomain of strain Bristol VP2 can efficiently encapsidate SA11 VP1. These results suggest that the VP2 NTD and dimer-forming subdomain play important, albeit non-specific, roles in both VP1 packaging and activation. When combined with previous work, the results of this study support the notion that the same VP2 regions that engage VP1 during activation are also involved in packaging the enzyme into the core.

Visualizing viral assemblies in a nanoscale biosphere.

We present a novel microfluidic platform to examine biological assemblies at high-resolution. We have engineered a functionalized chamber that serves as a "nanoscale biosphere" to capture and maintain rotavirus double-layered particles (DLPs) in a liquid environment. The chamber can be inserted into the column of a transmission electron microscope while being completely isolated from the vacuum system. This configuration allowed us to determine the structure of biological complexes at nanometer-resolution within a self-contained vessel. Images of DLPs were used to calculate the first 3D view of macromolecules in solution. We refer to this new fluidic visualization technology as in situ molecular microscopy.

In situ TEM of biological assemblies in liquid.

Researchers regularly use Transmission Electron Microscopes (TEMs) to examine biological entities and to assess new materials. Here, we describe an additional application for these instruments- viewing viral assemblies in a liquid environment. This exciting and novel method of visualizing biological structures utilizes a recently developed microfluidic-based specimen holder. Our video article demonstrates how to assemble and use a microfluidic holder to image liquid specimens within a TEM. In particular, we use simian rotavirus double-layered particles (DLPs) as our model system. We also describe steps to coat the surface of the liquid chamber with affinity biofilms that tether DLPs to the viewing window. This permits us to image assemblies in a manner that is suitable for 3D structure determination. Thus, we present a first glimpse of subviral particles in a native liquid environment.

Capturing RNA-dependent pathways for cryo-EM analysis.

Cryo-Electron Microscopy (EM) is a powerful technique to visualize biological processes at nanometer resolution. Structural studies of macromolecular assemblies are typically performed on individual complexes that are biochemically isolated from their cellular context. Here we present a molecular imaging platform to capture and view multiple components of cellular pathways within a functionally relevant framework. We utilized the bacterial protein synthesis machinery as a model system to develop our approach. By using modified Affinity Grid surfaces, we were able to recruit multiple protein assemblies bound to nascent strands of mRNA. The combined use of Affinity Capture technology and single particle electron microscopy provide the basis for visualizing RNA-dependent pathways in a remarkable new way.

Proliferation of antibiotic-producing bacteria and concomitant antibiotic production as the basis for the antibiotic activity of Jordan's red soils.

Anecdotes, both historical and recent, recount the curing of skin infections, including diaper rash, by using red soils from the Hashemite Kingdom of Jordan. Following inoculation of red soils isolated from geographically separate areas of Jordan, Micrococcus luteus and Staphylococcus aureus were rapidly killed. Over the 3-week incubation period, the number of specific types of antibiotic-producing bacteria increased, and high antimicrobial activity (MIC, approximately 10 microg/ml) was observed in methanol extracts of the inoculated red soils. Antibiotic-producing microorganisms whose numbers increased during incubation included actinomycetes, Lysobacter spp., and Bacillus spp. The actinomycetes produced actinomycin C(2) and actinomycin C(3). No myxobacteria or lytic bacteriophages with activity against either M. luteus or S. aureus were detected in either soil before or after inoculation and incubation. Although protozoa and amoebae were detected in the soils, the numbers were low and did not increase over the incubation period. These results suggest that the antibiotic activity of Jordan's red soils is due to the proliferation of antibiotic-producing bacteria.

Influence of hospital length of stay for heart failure on quality of care.

Adherence to treatment guidelines during hospital admissions for heart failure impacts readmissions and mortality. However, the relation between guideline adherence and heart failure hospital length of stay (LOS) has not been well studied. Whether quality of care delivered to patients with heart failure is impacted on by hospital LOS was assessed. Data were analyzed from 209 hospitals participating in the Get With the Guidelines heart failure program. From January 2005 to September 2006, a total of 36,078 admissions were recorded and stratified by a median heart failure hospitalization LOS of <5 or >or=5 days. Comparisons of baseline patient characteristics and quality measures were analyzed using generalized estimating equations. Patients with LOS >or=5 days were slightly older, more likely to be seen at a larger hospital, and had higher ejection fractions and increased rates of such co-morbidities as diabetes, anemia, renal insufficiency, and pulmonary disease. After adjustment, longer LOS was associated with an increased odds ratio (OR) per each additional day for providing discharge instructions (OR 1.027, 95% confidence interval [CI] 1.017 to 1.038) and left ventricular ejection fraction documentation (OR 1.049, 95% CI 1.031 to 1.067). However, LOS >or=5 days was independently associated with modestly decreased use of life-prolonging medications at hospital discharge for patients with left ventricular systolic dysfunction: angiotensin-converting enzyme inhibitors/angiotensin receptor blockers (OR 0.977, 95% CI 0.967 to 0.987) and beta blockers (OR 0.990, 95% CI 0.982 to 0.997). In conclusion, in Get With the Guidelines participating institutions, hospital LOS had only a modest influence on quality-of-care measures. Overall, excellent adherence to guideline-based medical therapy was observed, even in patients with a shorter hospital LOS.

Prospective randomized trial of wavefront-guided laser in situ keratomileusis with the CustomCornea and CustomVue laser systems.

PURPOSE: To compare visual function, safety, and higher-order aberrations (HOAs) after wavefront-guided laser in situ keratomileusis (LASIK) with the LadarVision CustomCornea (Alcon Laboratories, Inc.) and Star S4 CustomVue (Visx) laser systems. SETTING: John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Seventy-eight eyes of 39 patients with myopia with or without astigmatism were randomized for LASIK treatment in 1 eye with the CustomCornea laser; the other eye was treated with the CustomVue laser. Patients were followed for 6 months after surgery. The primary outcome measures were uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction, and changes in HOAs. RESULTS: At 6 months, the mean logarithm of the minimum angle of resolution (logMAR) UCVA was -0.0135 +/- 0.07 (SD) in the CustomVue group and 0.0417 +/- 0.12 in the CustomCornea group (P = .023). Eighty-eight percent of eyes in the CustomVue group had 20/20 or better UCVA compared with 67% in the CustomCornea group (P<.02). At 6 months, 91% of eyes in the CustomVue group and 79% in the CustomCornea group were within +/-0.50 diopter (D) of emmetropia (P<.1); 88% and 50%, respectively, were within +/-0.25 D (P<.001). Both platforms led to a small increase in total HOAs. The CustomVue system reduced trefoil and induced less of an increase in total HOAs, whereas the CustomCornea platform increased trefoil but induced less of an increase in spherical aberrations and coma. CONCLUSIONS: Both laser systems were effective, safe, and predictable. Wavefront-guided LASIK with the CustomVue system resulted in better visual acuity, with more eyes having 20/20 acuity than in the CustomCornea group.