Post-stroke kinematic analysis in rats reveals similar reaching abnormalities as humans.

A coordinated pattern of multi-muscle activation is essential to produce efficient reaching trajectories. Disruption of these coordinated activation patterns, termed synergies, is evident following stroke and results in reaching deficits; however, preclinical investigation of this phenomenon has been largely ignored. Furthermore, traditional outcome measures of post-stroke performance seldom distinguish between impairment restitution and compensatory movement strategies. We sought to address this by using kinematic analysis to characterize reaching movements and kinematic synergies of rats performing the Montoya staircase task, before and after ischemic stroke. Synergy was defined as the simultaneous movement of the wrist and other proximal forelimb joints (i.e. shoulder, elbow) during reaching. Following stroke, rats exhibited less individuation between joints, moving the affected limb more as a unit. Moreover, abnormal flexor synergy characterized by concurrent elbow flexion, shoulder adduction, and external rotation was evident. These abnormalities ultimately led to inefficient and unstable reaching trajectories, and decreased reaching performance (pellets retrieved). The observed reaching abnormalities in this preclinical stroke model are similar to those classically observed in humans. This highlights the potential of kinematic analysis to better align preclinical and clinical outcome measures, which is essential for developing future rehabilitation strategies following stroke.

Cyclosporin A-Mediated Activation of Endogenous Neural Precursor Cells Promotes Cognitive Recovery in a Mouse Model of Stroke.

Cognitive dysfunction following stroke significantly impacts quality of life and functional independance; yet, despite the prevalence and negative impact of cognitive deficits, post-stroke interventions almost exclusively target motor impairments. As a result, current treatment options are limited in their ability to promote post-stroke cognitive recovery. Cyclosporin A (CsA) has been previously shown to improve post-stroke functional recovery of sensorimotor deficits. Interestingly, CsA is a commonly used immunosuppressant and also acts directly on endogenous neural precursor cells (NPCs) in the neurogenic regions of the brain (the periventricular region and the dentate gyrus). The immunosuppressive and NPC activation effects are mediated by calcineurin-dependent and calcineurin-independent pathways, respectively. To develop a cognitive stroke model, focal bilateral lesions were induced in the medial prefrontal cortex (mPFC) of adult mice using endothelin-1. First, we characterized this stroke model in the acute and chronic phase, using problem-solving and memory-based cognitive tests. mPFC stroke resulted in early and persistent deficits in short-term memory, problem-solving and behavioral flexibility, without affecting anxiety. Second, we investigated the effects of acute and chronic CsA treatment on NPC activation, neuroprotection, and tissue damage. Acute CsA administration post-stroke increased the size of the NPC pool. There was no effect on neurodegeneration or lesion volume. Lastly, we looked at the effects of chronic CsA treatment on cognitive recovery. Long-term CsA administration promoted NPC migration toward the lesion site and rescued cognitive deficits to control levels. This study demonstrates that CsA treatment activates the NPC population, promotes migration of NPCs to the site of injury, and leads to improved cognitive recovery following long-term treatment.

Reconstructing Tone Sequences from Functional Magnetic Resonance Imaging Blood-Oxygen Level Dependent Responses within Human Primary Auditory Cortex.

Here we show that, using functional magnetic resonance imaging (fMRI) blood-oxygen level dependent (BOLD) responses in human primary auditory cortex, it is possible to reconstruct the sequence of tones that a person has been listening to over time. First, we characterized the tonotopic organization of each subject's auditory cortex by measuring auditory responses to randomized pure tone stimuli and modeling the frequency tuning of each fMRI voxel as a Gaussian in log frequency space. Then, we tested our model by examining its ability to work in reverse. Auditory responses were re-collected in the same subjects, except this time they listened to sequences of frequencies taken from simple songs (e.g., "Somewhere Over the Rainbow"). By finding the frequency that minimized the difference between the model's prediction of BOLD responses and actual BOLD responses, we were able to reconstruct tone sequences, with mean frequency estimation errors of half an octave or less, and little evidence of systematic biases.

Self-association motifs in the enteroaggregative Escherichia coli heat-resistant agglutinin 1.

The heat-resistant agglutinin 1 (Hra1) is an integral outer membrane protein found in strains of Escherichia coli that are exceptional colonizers. Hra1 from enteroaggregative E. coli strain 042 is sufficient to confer adherence to human epithelial cells and to cause bacterial autoaggregation. Hra1 is closely related to the Tia invasin, which also confers adherence, but not autoaggregation. Here, we have demonstrated that Hra1 mediates autoaggregation by self-association and we hypothesize that at least some surface-exposed amino acid sequences that are present in Hra1, but absent in Tia, represent autoaggregation motifs. We inserted FLAG tags along the length of Hra1 and used immune-dot blots to verify that four in silico-predicted outer loops were indeed surface exposed. In Hra1 we swapped nine candidate motifs in three of these loops, ranging from one to ten amino acids in length, to the corresponding sequences in Tia. Three of the motifs were required for Hra1-mediated autoaggregation. The database was searched for other surface proteins containing these motifs; the GGXWRDDXK motif was also present in a surface-exposed region of Rck, a Salmonella enterica serotype Typhimurium complement resistance protein. Cloning and site-specific mutagenesis demonstrated that Rck can confer weak, GGXWRDDXK-dependent autoaggregation by self-association. Hra1 and Rck appear to form heterologous associations and GGXWRDDXK is required on both molecules for Hra1-Rck association. However, a GGYWRDDLKE peptide was not sufficient to interfere with Hra1-mediated autoaggregation. In the present study, three autoaggregation motifs in an integral outer membrane protein have been identified and it was demonstrated that at least one of them works in the context of a different cell surface.

Assessing cognitive function following medial prefrontal stroke in the rat.

Cognitive impairments are prevalent following clinical stroke; however, preclinical research has focused almost exclusively on motor deficits. In order to conduct systematic evaluations into the nature of post-stroke cognitive dysfunction and recovery, it is crucial to develop focal stroke models that predominantly affect cognition while leaving motor function intact. Herein, we evaluated a range of cognitive functions 1-4 months following focal medial prefrontal cortex (mPFC) stroke using a battery of tests. Male Sprague-Dawley rats underwent focal ischemia induced in the mPFC using bilateral intracerebral injections of endothelin-1, or sham surgery. Cognitive function was assessed using an open field, several object recognition tests, attentional set-shifting, light-dark box, spontaneous alternation, Barnes maze, and win-shift/win-stay tests. Prefrontal cortex damage resulted in significant changes in object recognition function, behavioural flexibility, and anxiety-like behaviour, while spontaneous alternation and locomotor function remained intact. These deficits are similar to the cognitive deficits following stroke in humans. Our results suggest that this model may be useful for identifying and developing potential therapies for improving post-stroke cognitive dysfunction.

Population receptive field estimates of human auditory cortex.

Here we describe a method for measuring tonotopic maps and estimating bandwidth for voxels in human primary auditory cortex (PAC) using a modification of the population Receptive Field (pRF) model, developed for retinotopic mapping in visual cortex by Dumoulin and Wandell (2008). The pRF method reliably estimates tonotopic maps in the presence of acoustic scanner noise, and has two advantages over phase-encoding techniques. First, the stimulus design is flexible and need not be a frequency progression, thereby reducing biases due to habituation, expectation, and estimation artifacts, as well as reducing the effects of spatio-temporal BOLD nonlinearities. Second, the pRF method can provide estimates of bandwidth as a function of frequency. We find that bandwidth estimates are narrower for voxels within the PAC than in surrounding auditory responsive regions (non-PAC).

Voluntary forced use of the impaired limb following stroke facilitates functional recovery in the rat.

Constraint induced movement therapy (CIMT), which forces use of the impaired arm following stroke, improves functional recovery. The mechanisms underlying recovery are not well understood, necessitating further investigation into how rehabilitation may affect neuroplasticity using animal models. Animal motivation and stress make modelling CIMT in animals challenging. We have shown that following focal ischemia, voluntary forced use therapy using pet activity balls could engage the impaired forelimb and result in a modest acceleration in recovery. In this study, we investigated the effects of a more intensive appetitively motivated regimen that included task specific reaching exercises. Adult male Sprague Dawley rats were subjected to focal unilateral stroke using intracerebral injections of endothelin-1 or sham surgery. Three days later, stroke animals were assigned to daily rehabilitation or control therapy. Rehabilitation consisted of 30 min of generalized movement sessions in activity balls, followed by 30 min of voluntary task-specific movement using reaching boxes. Rats were tested weekly to measure forelimb deficit and recovery. After 30 days, animals were euthanized and tissue was examined for infarct volume, brain derived neurotrophic factor expression, and the presence of new neurons using doublecortin immunohistochemistry. Rehabilitation resulted in a significant acceleration of forelimb recovery in several tests, and a significant increase in the number of doublecortin-expressing cells. Furthermore, while the proportion of cells expressing BDNF in the peri-infarct region did not change, there was a shift in the cellular origin of expressed BDNF, resulting in significantly more non-neuronal, non-astrocytic BDNF, presumed to be of microglial origin.

Minimizing biases in estimating the reorganization of human visual areas with BOLD retinotopic mapping.

There is substantial interest in using functional magnetic resonance imaging (fMRI) retinotopic mapping techniques to examine reorganization of the occipital cortex after vision loss in humans and nonhuman primates. However, previous reports suggest that standard phase encoding and the more recent population Receptive Field (pRF) techniques give biased estimates of retinotopic maps near the boundaries of retinal or cortical scotomas. Here we examine the sources of this bias and show how it can be minimized with a simple modification of the pRF method. In normally sighted subjects, we measured fMRI responses to a stimulus simulating a foveal scotoma; we found that unbiased retinotopic map estimates can be obtained in early visual areas, as long as the pRF fitting algorithm takes the scotoma into account and a randomized "multifocal" stimulus sequence is used.

Cranberries and wild blueberries treated with gastrointestinal enzymes positively modify glutathione mechanisms in Caco-2 cells in vitro.

Beneficial health effects of cranberries (CBs) and wild blueberries (BBs), such as reduced levels of oxidative stress, have been demonstrated in feeding studies. These Vaccinium berries contain high levels of flavonoids; however, the bioavailability of flavonoids is generally low. We investigated the in vitro effects of these berries on intestinal cells, focusing on mitigating oxidative stress and associated reactive oxygen species (ROS). First, we simulated the passage of CB and BB through the gastrointestinal (GI) tract by treating berry homogenates to a battery of digestive enzymes. Then, Caco-2 cells, a model of small intestine epithelial uptake, were exposed to these homogenates for 60 min. Using a cell-free assay, we found that the antioxidant activity in CB homogenates was not affected by these enzymes, but that BB homogenates treated with gut enzymes had 43% lower free-radical quenching activity (P < 0.05). However, both of the enzyme-treated homogenates were still able to counteract the ROS-generating ability of H2O2 added exogenously to Caco-2 cells. Berry homogenates also increased mitochondrial metabolic rates at 60 min posttreatment, as measured by MTT assays. Enzyme-treated CB (but not BB) homogenates increased the levels of reduced glutathione (GSH) relative to oxidized glutathione (GSSG), a critical indicator of the cellular redox state (P < 0.05). Our data suggest that CBs do not lose their antioxidant ability when passing through the GI tract, and specifically, digested CB may serve to enhance cytoprotective effects in intestinal cells by reducing potential damage caused by free radicals and ROS derived from other food sources.

Animal models of post-ischemic forced use rehabilitation: methods, considerations, and limitations.

Many survivors of stroke experience arm impairments, which can severely impact their quality of life. Forcing use of the impaired arm appears to improve functional recovery in post-stroke hemiplegic patients, however the mechanisms underlying improved recovery remain unclear. Animal models of post-stroke rehabilitation could prove critical to investigating such mechanisms, however modeling forced use in animals has proven challenging. Potential problems associated with reported experimental models include variability between stroke methods, rehabilitation paradigms, and reported outcome measures. Herein, we provide an overview of commonly used stroke models, including advantages and disadvantages of each with respect to studying rehabilitation. We then review various forced use rehabilitation paradigms, and highlight potential difficulties and translational problems. Lastly, we discuss the variety of functional outcome measures described by experimental researchers. To conclude, we outline ongoing challenges faced by researchers, and the importance of translational communication. Many stroke patients rely critically on rehabilitation of post-stroke impairments, and continued effort toward progression of rehabilitative techniques is warranted to ensure best possible treatment of the devastating effects of stroke.

Adaptive radiotherapy for muscle-invasive bladder cancer: optimisation of plan sizes.

INTRODUCTION: Plan-of-the-day adaptive radiotherapy (ART) that has not been optimally designed may result in inefficient plan sizes. This can lead to unused plans, which may potentially reduce overall conformality. We compared two methods of individualising ART plan sizes for muscle-invasive bladder cancer to determine which provides a more balanced distribution of plan selections. METHODS: Twenty-seven previously treated patients had small, medium and large ART plans generated from CTV contours on the simulation CT and initial cone beam CTs (CBCT). In the original clinical method, the smallest plan was based on the smallest CTV, while the experimental method used the Boolean summation of the two smallest CTVs. The large plan was identical in both methods. The medium plans were created midway between small and large CTVs. Credentialed treatment staff performed plan selection clinically for the original plans and retrospectively for the experimental plans. RESULTS: A total of 646 CBCTs from 26 patients were included. The small, medium and large adaptive CTVs, and the conventional CTV, were used 29.7%, 45.4%, 22.0% and 2.9% of the time, respectively, compared to the previous 9.8%, 49.2%, 39.5% and 1.5%. The differences were significant between previous and new CTV (small), and CTV (large). CONCLUSIONS: The new design method resulted in the three adaptive CTV choices being selected more evenly, however, a reduction in a surrogate for normal tissue irradiation was not observed.

Stimulus-specific adaptation in specialized neurons in the inferior colliculus of the big brown bat, Eptesicus fuscus.

The inferior colliculus (IC) of the big brown bat (Eptesicus fuscus) contains specialized neurons that respond exclusively to highly specific spectrotemporal patterns such as sinusoidally frequency modulated (SFM) signals or directional frequency modulated sweeps (FM). Other specialized cells with I-shaped frequency response areas (FRAs) are tuned to very narrow frequency bands (1-2 kHz) in an amplitude-tolerant manner. In contrast, non-specialized neurons respond to any stimulus with energy in their frequency response area. IC neurons in several mammalian species, including bats, demonstrate stimulus-specific adaptation (SSA), a reduction in response to a high-probability stimulus. To evaluate the relation between stimulus selectivity and SSA, we presented sounds using an oddball stimulus paradigm and recorded extracellular responses of IC neurons. SFM-selective cells (n = 10), FM-selective cells (n = 7), and cells with I-shaped FRAs (n = 13) did not show SSA under any of the conditions tested (NSSI = 0.009, 0.033, 0.020 respectively). However, non-specialized neurons (n = 52) exhibited various levels of SSA (NSSI = 0.163), with a subset of these cells displaying strong adaptation. These findings suggest that SSA is not a ubiquitous characteristic of all neurons in the bat IC, but is present only in a subset of non-specialized neurons.

Impact of post-therapy positron emission tomography on prognostic stratification and surveillance after chemoradiotherapy for cervical cancer.

BACKGROUND: A study was undertaken to investigate the detection of relapse and survival outcomes in patients with cervical cancer treated with curative intent chemoradiotherapy, and evaluated with a post-therapy (18) F-fluorodeoxyglucose positron emission tomography (FDG-PET) scan. METHODS: Between January 2002 and June 2007, 105 consecutive patients were prospectively enrolled into a registry study designed to assess outcomes of chemoradiotherapy. A FDG-PET scan was performed between 3 and 12 months (median, 4.9 months) post-treatment at clinician discretion. Tumor response was graded as complete metabolic response, partial metabolic response, or progressive metabolic disease. RESULTS: Median follow-up was 36 months. At post-therapy FDG-PET, 73 (70%) patients had complete metabolic response, 10 (9%) had partial metabolic response, and 22 (21%) had progressive metabolic disease. Overall survival at 3 years was 77% in all patients, and 95% for those with complete metabolic response. On multivariate analysis, complete metabolic response (P < .0001) and pretreatment tumor volume (P = .041) were strong predictors for overall survival. The number of involved lymph nodes (P < .005) and International Federation of Gynecology and Obstetrics stage (P = .04) were predictive of relapse-free survival. In total, 18 patients relapsed at a single site, and 13 underwent salvage, with a 3-year survival of 67%. Patients with complete metabolic response had a distant failure rate 36-fold less than those with partial metabolic response (P < .0001). After complete metabolic response, only 1 patient (1.6%) relapsed without symptoms and was detected through physical examination. CONCLUSIONS: The presence of a complete metabolic response at post-therapy FDG-PET is a powerful predictor for survival after chemoradiation. The very low rate of recurrence in patients with a complete metabolic response justifies a conservative follow-up approach for these patients, because relapse is usually symptomatic and not detected by routine clinical review.