Active stiffness of the ankle in response to inertial and elastic loads.

Effective stiffness of the musculoskeletal system was examined as a function of the characteristics of an external load. Thirteen healthy subjects provided active contraction of the ankle plantarflexion musculature in a neutral ankle posture to support an external load. Musculoskeletal stiffness was computed from kinetic data recorded in response to dorsiflexion/plantarflexion perturbations. Ankle dynamics were recorded while supporting external loads of 19 and 38 kg with and without antagonistic co-contraction. External loads were applied using pure gravitational mass. In separate trials external loads were applied from stretch of steel springs in parallel with the plantarflexion musculature that also provided added parallel stiffness to the system. Adding external stiffness of 4.9 and 8.1 kN/m surprisingly failed to significantly change the stiffness of the ankle-plus-spring system. This suggests contributions from intrinsic muscle stiffness and reflex stiffness declined in response to added external stiffness. This could not be explained by load magnitudes, ankle postures, or co-activation as these were similar between the inertial and elastic loading conditions. However, non-linear parametric analyses suggest mean intrinsic stiffness of 35.5 kN/m and reflex gain of 11.6 kN/m with a constant reflex delay of 70 ms accurately described the empirical results. The phase response between the mechanical dynamics of the musculoskeletal system and delayed neuromotor feedback combine to provide robust control of system behavior.     

New insights into Alzheimer's disease progression: a combined TMS and structural MRI study

Background:

Combination of structural and functional data of the human brain can provide detailed information of neurodegenerative diseases and the influence of the disease on various local cortical areas.

Methodology and Principal Findings:

To examine the relationship between structure and function of the brain the cortical thickness based on structural magnetic resonance images and motor cortex excitability assessed with transcranial magnetic stimulation were correlated in Alzheimer''s disease (AD) and mild cognitive impairment (MCI) patients as well as in age-matched healthy controls. Motor cortex excitability correlated negatively with cortical thickness on the sensorimotor cortex, the precuneus and the cuneus but the strength of the correlation varied between the study groups. On the sensorimotor cortex the correlation was significant only in MCI subjects. On the precuneus and cuneus the correlation was significant both in AD and MCI subjects. In healthy controls the motor cortex excitability did not correlate with the cortical thickness.

Conclusions:

In healthy subjects the motor cortex excitability is not dependent on the cortical thickness, whereas in neurodegenerative diseases the cortical thinning is related to weaker cortical excitability, especially on the precuneus and cuneus. However, in AD subjects there seems to be a protective mechanism of hyperexcitability on the sensorimotor cortex counteracting the prominent loss of cortical volume since the motor cortex excitability did not correlate with the cortical thickness. Such protective mechanism was not found on the precuneus or cuneus nor in the MCI subjects. Therefore, our results indicate that the progression of the disease proceeds with different dynamics in the structure and function of neuronal circuits from normal conditions via MCI to AD.     

A homogeneous HTRF assay for the identification of inhibitors of the TWEAK-Fn14 protein interaction

The TWEAK-Fn14 pathway is upregulated in models of inflammation, autoimmune diseases, and cancer. Both TWEAK and Fn14 show increased expression also in the CNS in response to different stimuli, particularly astrocytes, microglia, and neurons, leading to activation of NF-κB and release of proinflammatory cytokines. Although neutralizing antibodies against these proteins have been shown to have therapeutic efficacy in animal models of inflammation, no small-molecule therapeutics are yet available. Here, we describe the development of a novel homogeneous time-resolved fluorescence (HTRF)-based screening assay together with several counterassays for the identification of small-molecule inhibitors of this protein-protein interaction. Recombinant HIS-TWEAK and Fn14-Fc proteins as well as FLAG-TWEAK and Fn14-FLAG proteins and an anti-Fn14 antibody were used to establish and validate these assays and to screen a library of 60 000 compounds. Two HTRF counterassays with unrelated proteins in the same assay format, an antiaggregation assay and a redox assay, were applied to filter out potential false-positive compounds. The novel assay and associated screening cascade should be useful for the discovery of small-molecule inhibitors of the TWEAK-Fn14 protein interaction.     

Diet‐induced early‐stage atherosclerosis in baboons: Lipoproteins,atherogenesis, and arterial compliance

Background

The purpose of this study was to determine whether dietary manipulation can reliably induce early‐stage atherosclerosis and clinically relevant changes in vascular function in an established, well‐characterized non‐human primate model.

Methods

We fed 112 baboons a high‐cholesterol, high‐fat challenge diet for two years. We assayed circulating biomarkers of cardiovascular disease (CVD) risk, at 0, 7, and 104 weeks into the challenge; assessed arterial compliance noninvasively at 104 weeks; and measured atherosclerotic lesions in three major arteries at necropsy.

Results

We observed evidence of atherosclerosis in all but one baboon fed the two‐year challenge diet. CVD risk biomarkers, the prevalence, size, and complexity of arterial lesions, plus consequent arterial stiffness, were increased in comparison with dietary control animals.

Conclusions

Feeding baboons a high‐cholesterol, high‐fat diet for two years reliably induces atherosclerosis, with risk factor profiles, arterial lesions, and changes in vascular function also seen in humans.     

EEG Responses to TMS Are Sensitive to Changes in the Perturbation Parameters and Repeatable over Time

Background

High-density electroencephalography (hd-EEG) combined with transcranial magnetic stimulation (TMS) provides a direct and non-invasive measure of cortical excitability and connectivity in humans and may be employed to track over time pathological alterations, plastic changes and therapy-induced modifications in cortical circuits. However, the diagnostic/monitoring applications of this technique would be limited to the extent that TMS-evoked potentials are either stereotypical (non-sensitive) or random (non-repeatable) responses. Here, we used controlled changes in the stimulation parameters (site, intensity, and angle of stimulation) and repeated longitudinal measurements (same day and one week apart) to evaluate the sensitivity and repeatability of TMS/hd-EEG potentials.

Methodology/Principal Findings

In 10 volunteers, we performed 92 single-subject comparisons to evaluate the similarities/differences between pairs of TMS-evoked potentials recorded in the same/different stimulation conditions. For each pairwise comparison, we used non-parametric statistics to calculate a Divergence Index (DI), i.e., the percentage of samples that differed significantly, considering all scalp locations and the entire post-stimulus period. A receiver operating characteristic analysis showed that it was possible to find an optimal DI threshold of 1.67%, yielding 96.7% overall accuracy of TMS/hd-EEG in detecting whether a change in the perturbation parameters occurred or not.

Conclusions/Significance

These results demonstrate that the EEG responses to TMS essentially reflect deterministic properties of the stimulated neuronal circuits as opposed to stereotypical responses or uncontrolled variability. To the extent that TMS-evoked potentials are sensitive to changes and repeatable over time, they may be employed to detect longitudinal changes in the state of cortical circuits.     

Stimulus Set Meaningfulness and Neurophysiological Differentiation: A Functional Magnetic Resonance Imaging Study

A meaningful set of stimuli, such as a sequence of frames from a movie, triggers a set of different experiences. By contrast, a meaningless set of stimuli, such as a sequence of ‘TV noise’ frames, triggers always the same experience—of seeing ‘TV noise’—even though the stimuli themselves are as different from each other as the movie frames. We reasoned that the differentiation of cortical responses underlying the subject’s experiences, as measured by Lempel-Ziv complexity (incompressibility) of functional MRI images, should reflect the overall meaningfulness of a set of stimuli for the subject, rather than differences among the stimuli. We tested this hypothesis by quantifying the differentiation of brain activity patterns in response to a movie sequence, to the same movie scrambled in time, and to ‘TV noise’, where the pixels from each movie frame were scrambled in space. While overall cortical activation was strong and widespread in all conditions, the differentiation (Lempel-Ziv complexity) of brain activation patterns was correlated with the meaningfulness of the stimulus set, being highest in the movie condition, intermediate in the scrambled movie condition, and minimal for ‘TV noise’. Stimulus set meaningfulness was also associated with higher information integration among cortical regions. These results suggest that the differentiation of neural responses can be used to assess the meaningfulness of a given set of stimuli for a given subject, without the need to identify the features and categories that are relevant to the subject, nor the precise location of selective neural responses.     

Identification of candidate genes encoding an LDL-C QTL in baboons

Cardiovascular disease (CVD) is the leading cause of death in developed countries, and dyslipidemia is a major risk factor for CVD. We previously identified a cluster of quantitative trait loci (QTL) on baboon chromosome 11 for multiple, related quantitative traits for serum LDL-cholesterol (LDL-C). Here we report differentially regulated hepatic genes encoding an LDL-C QTL that influences LDL-C levels in baboons. We performed hepatic whole-genome expression profiling for LDL-C-discordant baboons fed a high-cholesterol, high-fat (HCHF) diet for seven weeks. We detected expression of 117 genes within the QTL 2-LOD support interval. Three genes were differentially expressed in low LDL-C responders and 8 in high LDL-C responders in response to a HCHF diet. Seven genes (ACVR1B, CALCOCO1, DGKA, ERBB3, KRT73, MYL6B, TENC1) showed discordant expression between low and high LDL-C responders. To prioritize candidate genes, we integrated miRNA and mRNA expression profiles using network tools and found that four candidates (ACVR1B, DGKA, ERBB3, TENC1) were miRNA targets and that the miRNAs were inversely expressed to the target genes. Candidate gene expression was validated using QRT-PCR and Western blotting. This study reveals candidate genes that influence variation in LDL-C in baboons and potential genetic mechanisms for further investigation.     

Differential microRNA response to a high-cholesterol, high-fat diet in livers of low and high LDL-C baboons

ABSTRACT: BACKGROUND: Dysregulation of microRNA (miRNA) expression has been implicated in molecular geneticevents leading to the progression and development of atherosclerosis. We hypothesized thatmiRNA expression profiles differ between baboons with low and high serum low-densitylipoprotein cholesterol (LDL-C) concentrations in response to diet, and that a subset of thesemiRNAs regulate genes relevant to dyslipidemia and risk of atherosclerosis. RESULTS: Using Next Generation Illumina sequencing methods, we sequenced hepatic small RNAlibraries from baboons differing in their LDL-C response to a high-cholesterol, high-fat(HCHF) challenge diet (low LDL-C, n = 3; high LDL-C, n = 3), resulting in 517 baboonmiRNAs: 490 were identical to human miRNAs and 27 were novel. We compared miRNAexpression profiles from liver biopsies collected before and after the challenge diet andobserved that HCHF diet elicited expression of more miRNAs compared to baseline (chow)diet for both low and high LDL-C baboons. Eighteen miRNAs exhibited differentialexpression in response to HCHF diet in high LDL-C baboons compared to 10 miRNAs in low LDL-C baboons. We used TargetScan/Base tools to predict putative miRNA targets;miRNAs expressed in high LDL-C baboons had significantly more gene targets thanmiRNAs expressed in low LDL-C responders. Further, we identified miRNA isomers andother non-coding RNAs that were differentially expressed in response to the challenge diet inboth high LDL-C and low LDL-C baboons. CONCLUSIONS: We sequenced and annotated baboon liver miRNAs from low LDL-C and high LDL-Cresponders using high coverage Next Gen sequencing methods, determined expressionchanges in response to a HCHF diet challenge, and predicted target genes regulated by thedifferentially expressed miRNAs. The identified miRNAs will enrich the database for noncodingsmall RNAs including the extent of variation in these sequences. Further, weidentified other small non-coding RNAs differentially expressed in response to diet. Ourdiscovery of differentially expressed baboon miRNAs in response to a HCHF diet challengethat differ by LDL-C phenotype is a fundamental step in understating the role of non-codingRNAs in dyslipidemia.