Time Course Of Response Research Articles

Visual Working Memory of Chinese Characters and Expertise: The Expert's Memory Advantage Is Based on Long-Term Knowledge of Visual Word Forms.

People unfamiliar with Chinese characters show poorer visual working memory (VWM) performance for Chinese characters than do literates in Chinese. In a series of experiments, we investigated the reasons for this expertise advantage. Experiments 1 and 2 showed that the advantage of Chinese literates does not transfer to novel material. Experts had similar resolution as novices for material outside of their field of expertise, and the memory of novices and experts did not differ when detecting a big change, e.g., when a character’s color was changed. Memorizing appears to function as a rather abstract representation of word forms because memory for characters’ fonts was poor independently of expertise (Experiment 3), though still visual. Distractors that were highly similar conceptually did not increase memory errors, but visually similar distractors impaired memory (Experiment 4). We hypothesized that literates in Chinese represent characters in VWM as tokens of visual word forms made available by long-term memory. In Experiment 5, we provided novices with visual word form knowledge. Participants subsequently performed a change detection task with trained and novel characters in a functional magnetic resonance experiment. We analyzed set size- and training-dependent effects in the intraparietal sulcus (IPS) and the visual word form area. VWM for trained characters was better than for novel characters. Neural activity increased with set size and at a slower rate for trained than for novel characters. All conditions approached the same maximum, but novel characters reached the maximum at a smaller set size than trained characters. The time course of the bold response depended on set size and knowledge status. Starting from the same initial maximum, neural activity at small set sizes returned to baseline more quickly for trained characters than for novel characters. Additionally, high performers showed generally more neural activity in the IPS than low performers. We conclude that experts’ better performance in working memory (WM) is caused by the availability of visual long-term representations (word form types) that allow a sparse representation of the perceived stimuli and make even small changes big because they cause a type change that is easily detected.

Drug Choices and Advancements for Managing Depression in Parkinson's Disease.

Depression is a frequent non-motor symptom of Parkinson’s disease (PD), and may even precede the onset of motor symptoms of parkinsonism. Beyond its negative influence on mood, depression in PD is frequently associated with other neuropsychiatric symptoms and with late-stage complications such as dementia. Despite its profound impact on the quality of life and cognitive functioning in PD, depression in PD is often under-recognized and poorly treated.Pathophysiological studies demonstrated that depression in PD is associated with global dysfunction of interactions between discrete brain areas rather than focal structural or functional abnormalities, and that it is sustained by pathological changes of several neurotransmitter/receptor complexes.In general, all traditional antidepressants and some dopamine agonists have been found to be safe and well-tolerated to treat depressive symptoms in PD, despite initial warning on worsening of parkinsonism. Available data suggest that the time-course of response differs among antidepressants. Efficacy results from clinical trials with antidepressant in PD are, however, rather uncertain, although pooled analysis suggests a moderate benefit. Several issues may critically impact the results of clinical trials with antidepressants in PD, including the correct psychiatric diagnosis, the overlap of symptoms between depression and PD, and the selection of appropriate end-points and rating scales.

Cryolipolysis: Clinical Best Practices and Other Nonclinical Considerations.

Cryolipolysis is a nonsurgical body contouring procedure that involves cooling of fat cells to induce lipolysis while sparing surrounding structures. Plastic surgery practices are increasingly incorporating noninvasive aesthetic procedures (eg, cryolipolysis, fillers, radiofrequency, ultrasound) to offer their patients a wider range of aesthetic treatment options. Here, we report insights from 8 plastic surgeons with regard to cryolipolysis best practices from a clinical perspective and the impact of integrating this noninvasive body contouring procedure into a plastic surgery practice. The authors prefer cryolipolysis over liposuction for patients who are not amenable to surgery or those who desire to avoid downtime, also taking into consideration body mass index, skin laxity, comorbidities, and risk of contour irregularities. Patient counseling is critical for setting realistic expectations regarding outcomes and should focus on the efficacy of cryolipolysis, individual variability in results, potential side effects, time course of treatment response, and the need for multiple treatment cycles. Strategies for reaching new patients and expanding services among current cryolipolysis patients are discussed.

P282 Navigated repetitive transcranial magnetic stimulation at 10 Hz in chronic neuropathic pain – time course, duration and predictors of response

P282 Navigated repetitive transcranial magnetic stimulation at 10 Hz in chronic neuropathic pain – time course, duration and predictors of response

Temporal responses of conserved miRNAs to drought and their associations with drought tolerance and productivity in rice

BackgroundPlant miRNAs play crucial roles in responses to drought and developmental processes. It is essential to understand the association of miRNAs with drought-tolerance (DT), as well as their impacts on growth, development, and reproduction (GDP). This will facilitate our utilization of rice miRNAs in breeding.ResultsIn this study, we investigated the time course of miRNA responses to a long-term drought among six rice genotypes by high-throughput sequencing. In total, 354 conserved miRNAs were drought responsive, representing obvious genotype- and stage-dependent patterns. The drought-responsive miRNAs (DRMs) formed complex regulatory network via their coexpression and direct/indirect impacts on the rice transcriptome. Based on correlation analyses, 211 DRMs were predicted to be associated with DT and/or GDP. Noticeably, 14.2% DRMs were inversely correlated with DT and GDP. In addition, 9 pairs of mature miRNAs, each derived from the same pre-miRNAs, were predicted to have opposite roles in regulating DT and GDP. This suggests a potential yield penalty if an inappropriate miRNA/pre-miRNA is utilized. miRNAs have profound impacts on the rice transcriptome reflected by great number of correlated drought-responsive genes. By regulating these genes, a miRNA could activate diverse biological processes and metabolic pathways to adapt to drought and have an influence on its GDP.ConclusionBased on the temporal pattern of miRNAs in response to drought, we have described the complex network between DRMs. Potential associations of DRMs with DT and/or GDP were disclosed. This knowledge provides valuable information for a better understanding in the roles of miRNAs play in rice DT and/or GDP, which can facilitate our utilization of miRNA in breeding.

GABAA Receptor β2E155 Residue Located at the Agonist-Binding Site Is Involved in the Receptor Gating.

GABAA receptors (GABAARs) play a crucial role in mediating inhibition in the adult brain. In spite of progress in describing (mainly) the static structures of this receptor, the molecular mechanisms underlying its activation remain unclear. It is known that in the α1β2γ2L receptors, the mutation of the β2E155 residue, at the orthosteric binding site, strongly impairs the receptor activation, but the molecular and kinetic mechanisms of this effect remain elusive. Herein, we investigated the impact of the β2E155C mutation on binding and gating of the α1β2γ2L receptor. To this end, we combined the macroscopic and single-channel analysis, the use of different agonists [GABA and muscimol (MSC)] and flurazepam (FLU) as a modulator. As expected, the β2E155C mutation caused a vast right shift of the dose–response (for GABA and MSC) and, additionally, dramatic changes in the time course of current responses, indicative of alterations in gating. Mutated receptors showed reduced maximum open probability and enhanced receptor spontaneous activity. Model simulations for macroscopic currents revealed that the primary effect of the mutation was the downregulation of the preactivation (flipping) rate. Experiments with MSC and FLU further confirmed a reduction in the preactivation rate. Our single-channel analysis revealed the mutation impact mainly on the second component in the shut times distributions. Based on model simulations, this finding further confirms that this mutation affects mostly the preactivation transition, supporting thus the macroscopic data. Altogether, we provide new evidence that the β2E155 residue is involved in both binding and gating (primarily preactivation).

A real-time marker of object-based attention in the human brain. A possible component of a “gate-keeping mechanism” performing late attentional selection in the Ventro-Lateral Prefrontal Cortex

The decision to process an incoming stimulus attentively - and to trigger a follow-up cascade of high-level processes - is strategic for the human brain as it becomes transiently unavailable to subsequent stimulus processing. In this study, we set to identify brain networks that carry out such evaluations. We therefore assessed the time-course of neural responses with intracerebral EEG in human patients during an attentional reading task, contrasting to-be-attended vs. to-be-ignored items. We measured High-Frequency Activity [50–150 ​Hz] as a proxy of population-level spiking activity and we identified a crucial component of a Gate-Keeping Mechanism bilateral in the mid-Ventro-Lateral Prefrontal Cortex (VLPFC), at the interplay of the Ventral and Dorsal Attention Networks, that selectively reacts before domain specialized cortical regions that engage in full stimulus analysis according to task demands.

Development of an in vitro system and model-based translational framework to assess haemolysis risk following intravenous abuse of medications containing polyethylene oxide

Multiple cases of potentially life-threatening thrombotic microangiopathy (TMA) have resulted from intravenous abuse of medications containing polyethylene oxide (PEO), most often Opana ER (oxymorphone hydrochloride extended release). No validated models are available to assess the risk of TMA with different formulations and extraction methods following intravenous abuse. We have developed an in vitro system that involves passing pooled blood containing the excipient of interest through a syringe needle and assessing haemolysis via haemoglobin release. Haemolysis is induced by high shear stress caused by the flow of blood containing PEO through a narrow-bore syringe needle, recapitulating the mechanism in small blood vessels. Using the in vitro system, we demonstrate that high-molecular-weight PEO (>1 MDa) induces haemolysis in a concentration-dependent manner under flowing but not static conditions. We use data from the in vitro system and published in vivo data to predict the time course of the haemolytic response in vivo via a pharmacometric model. The in vitro system is a novel method for investigating factors influencing PEO-induced haemolysis. In combination with our model-based translational framework, the in vitro system allows straightforward assessment of the haemolytic potential of PEO-containing medications, and may find application in gauging TMA risk following intravenous abuse.

Autistic traits modulate cortical responses to affective but not discriminative touch.

The sense of touch is primarily considered a discriminative and exteroceptive sense, facilitating the detection, manipulation and exploration of objects, via an array of low-threshold mechanoreceptors and fast conducting A-beta (Aβ) afferents. However, a class of unmyelinated, low-threshold mechanoreceptors identified in the hairy skin of mammals have been proposed to constitute a second, anatomically distinct system coding the affective qualities of touch. Unlike Aβs, which increase their firing rate linearly with the velocity of a stimulus moving across their receptive field, the response of these C-tactile afferents (CTs) is described by an inverted 'U' curve fit, responding optimally to a skin temperature stimulus moving at between 1 and 10cm/s. Given the distinct velocity tuning of these fast and slow touch fibres, here we used event-related potentials to compare the time course of neural responses to 1st (fast) and 2nd (slow) touch systems. We identified a higher amplitude P300 in response to fast, Aβ-targeted, versus slow CT-targeted, stroking touch. In contrast, we identified a previously described, C-fibre specific, ultra-late potential (ULP) associated with CT-targeted input. Of special note as regards the function of CTs is that the amplitude of the ULP was negatively correlated with self-reported levels of autistic traits, which is consistent with the hypothesized affective and social significance of this response. Taken together, these findings provide further support for distinct discriminative and affective touch systems and suggests the temporal resolution of EEG provides an as yet underutilized tool for exploring individual differences in response sensitivity to CT-targeted touch.

Early and later life environmental enrichment affect specific antibody responses and blood leukocyte subpopulations in pigs

This study addressed the impact of early and later life environmental enrichment, and their combination, on specific antibody responses and peripheral blood leukocyte subpopulations in pigs. Pigs were kept in either barren (B1) or enriched (E1) housing from birth, and half of the pigs switched to barren or enriched housing on day 47, resulting in four treatment combinations: B1B2, B1E2, E1B2, E1E2). Pigs were immunized with keyhole limpet hemocyanin-conjugated trinitrophenyl (KLH-TNP) on day 74 and 109 to induce primary and secondary antibody responses. Blood samples were taken weekly until day 130, and IgM and IgG antibody responses were measured. Leukocyte subpopulations were measured on day 74 and 130. Time course of the antibody responses was not affected by housing. Early life enrichment increased the IgG response to KLH, particularly the primary one. At day 74 the relative frequency of lymphocytes, DC and SLA-II expression on monocytes were higher in E1 pigs, whereas the percentage of granulocytes tended to be lower in E1 pigs at day 74. Early life enrichment increased the SLA-II expression on monocytes, the granulocyte to lymphocyte ratio, and tended to increase the percentage of granulocytes, but tended to decrease the percentage of monocytes at day 130. Later life enrichment reduced percentages of CD4+CD8α+ T cells before and after immunization and the SLA-II expression on monocytes at day 74, the percentage of granulocytes and the granulocyte to lymphocyte ratio at day 130. Notably, early and later life housing interacted in their effects on several immune parameters. KLH-IgM responses (both primary and secondary) were affected by the interaction between early and later life housing. IgM titers were higher for B1B2 than for E1E2, with the switched animals (B1E2 and E1B2) moving towards the titers of the animals kept in their later life environment from birth onwards. At day 130 the percentage of gamma delta T cells, CD8α+ cytotoxic T cells and DC were not different between pigs kept in B1B2 and E1E2, but there was a clear impact of the switch in housing conditions, particularly for the pigs that changed from barren to enriched housing. We also found effects of coping style (personality) and sex on some immune parameters. In conclusion, both early life and later life enrichment, and, notably a switch in housing conditions influenced specific antibodies and leukocyte subpopulations in pigs. The current study implies that the early life history of animals and the (mis)match with their current environment could thus be of major importance for their immune system. Further research is needed to investigate potential consequences for the pigs’ health.

Functional asymmetry of local connections in V1 and its impact on orientation tuning

Abstract Classic models of primary visual cortex (V1) reproduce orientation selectivity with sufficiently high accuracy. However, they are unable to explain phenomena that are widely studied experimentally: a single cell carry additional information in response waveform and oscillations. Among the other effects not reproduced by the classic models are changes in the peak orientation during the response time course and shifts induced by inhibition blockage. We suppose that the main reason for such a failure is assumption that spatial distribution of short-range horizontal connections in V1 is symmetric. In the present study, we provide an experimental evidence that such symmetry is not the case in higher mammals. Based on this evidence we improved the classical model of V1 by replacing the symmetric spatial distribution to another one revealed from experiments. The improved model is able to reproduce orientation shifts and transmission of additional information. This means that the asymmetry shall take into consideration when simulating V1.

Functional connectivity of the irritative zone identified by electrical source imaging, and EEG-correlated fMRI analyses

The irritative zone - the area generating epileptic spikes - can be studied non-invasively during the interictal period using Electrical Source Imaging (ESI) and simultaneous electroencephalography-functional magnetic resonance imaging (EEG-fMRI). Although the techniques yield results which may overlap spatially, differences in spatial localization of the irritative zone within the same patient are consistently observed. To investigate this discrepancy, we used Blood Oxygenation Level Dependent (BOLD) functional connectivity measures to examine the underlying relationship between ESI and EEG-fMRI findings. Fifteen patients (age 20-54), who underwent presurgical epilepsy investigation, were scanned using a single-session resting-state EEG-fMRI protocol. Structural MRI was used to obtain the electrode localisation of a high-density 64-channel EEG cap. Electrical generators of interictal epileptiform discharges were obtained using a distributed local autoregressive average (LAURA) algorithm as implemented in Cartool EEG software. BOLD activations were obtained using both spike-related and voltage-map EEG-fMRI analysis. The global maxima of each method were used to investigate the temporal relationship of BOLD time courses and to assess the spatial similarity using the Dice similarity index between functional connectivity maps. ESI, voltage-map and spike-related EEG-fMRI methods identified peaks in 15 (100%), 13 (67%) and 8 (53%) of the 15 patients, respectively. For all methods, maxima were localised within the same lobe, but differed in sub-lobar localisation, with a median distance of 22.8mm between the highest peak for each method. The functional connectivity analysis showed that the temporal correlation between maxima only explained 38% of the variance between the time course of the BOLD response at the maxima. The mean Dice similarity index between seed-voxel functional connectivity maps showed poor spatial agreement. Non-invasive methods for the localisation of the irritative zone have distinct spatial and temporal sensitivity to different aspects of the local cortical network involved in the generation of interictal epileptiform discharges.

Role of Pyroptosis in Traumatic Brain and Spinal Cord Injuries.

Central nervous system (CNS) trauma, including traumatic brain injury (TBI) and spinal cord injury (SCI), remains a leading cause for morbidity and mortality worldwide. Past research has shown that cell death plays a critical role in the pathophysiology of CNS injuries. More recently, pyroptosis has been identified as a form of programmed inflammatory cell death, and it is a unique form of cell death in various aspects. Mechanistically, pyroptosis can be categorized into canonical (mediated by caspase-1) and non-canonical (mediated by caspase-4/5/11). In canonical pyroptosis, Nod-like receptors (NLRs) inflammasomes play a critical role, and their activation promotes the maturation and secretion of the inflammatory cytokines interleukin-1β/18 (IL-1β/18), cleavage of gasdermin D (GSDMD), and ultimately pyroptotic cell death. Despite a plethora of new knowledge regarding pyroptosis, detailed understanding of how pyroptosis is involved in CNS injuries and possible ways to improve clinical outcomes following CNS injuries remain elusive. This review discusses the current knowledge on how pyroptosis is involved in CNS injuries, focusing on new discoveries regarding how pyroptosis activation occurs, differences between CNS cell types following injury, time-course of inflammatory responses, and key regulatory steps of pyroptosis. In addition, we highlight various investigational agents that are capable of regulating key steps in pyroptotic cell death, and we discuss how these agents may be used as therapies to improve outcomes following CNS trauma.

Whole body vibration elicits differential immune and metabolic responses in obese and normal weight individuals

Traditional aerobic exercise reduces the risk of developing chronic diseases by inducing immune, metabolic, and myokine responses. Following traditional exercise, both the magnitude and time-course of these beneficial responses are different between obese compared to normal weight individuals. Although obesity may affect the ability to engage in traditional exercise, whole body vibration (WBV) has emerged as a more tolerable form of exercise . The impact of WBV on immune, metabolic, and myokine responses as well as differences between normal weight and obese individuals, however, is unknown. PurposeTo determine if WBV elicits differential magnitudes and time-courses of immune, metabolic, and myokine responses between obese and normal weight individuals. Methods21 participants [Obese (OB): n = 11, Age: 33 ± 4 y, percent body fat (%BF): 39.1 ± 2.4% & Normal weight (NW) n = 10, Age: 28 ± 8 y, %BF: 17.4 ± 2.1%] engaged in 10 cycles of WBV exercise [1 cycle = 1 min of vibration followed by 30 s of rest]. Blood samples were collected pre-WBV (PRE), immediately (POST), 3 h (3H), and 24 h (24H) post-WBV and analyzed for leukocytes, insulin, glucose, and myokines (IL-6, decorin, myostatin). ResultsThe peak (3H) percent change in neutrophil counts (OB: 13.9 ± 17.4 vs. NW: 47.2 ± 6.2%Δ; p = 0.007) was different between groups. The percent change in neutrophil percentages was increased in NW (POST: -1.6 ± 2.0 vs. 3H: 13.0 ± 7.2%Δ, p = 0.019) but not OB (p > 0.05). HOMA β-cell function was increased at 24H (PRE: 83.4 ± 5.4 vs. 24H: 131.0 ± 14.1%; p = 0.013) in NW and was not altered in OB (p > 0.05). PRE IL-6 was greater in OB compared to NW (OB: 2.7 ± 0.6 vs. NW: 0.6 ± 0.1 pg/mL; p = 0.011); however, the percent change from PRE to peak (3H) was greater in NW (OB: 148.1 ± 47.9 vs. NW: 1277.9 ± 597.6 %Δ; p = 0.035). Creatine kinase, decorin, and myostatin were not significantly altered in either group (p > 0.05). ConclusionTaken together, these data suggest that acute whole body vibration elicits favorable immune, metabolic, and myokine responses and that these responses differ between obese and normal weight individuals.

Physiological assessment of high glucose neurotoxicity in mouse and rat retinal explants.

One of the tissues of the central nervous system most affected by diabetes is the retina. Despite a growing understanding of the biochemical processes involved in glucose toxicity, little is known about the physiological consequences of chronic high glucose (HG) on individual neurons and neuronal circuits. Electroretinogram recordings suggest that retinal bipolar cells (BCs), which filter and transmit photoreceptor output to the inner retina, are among the first cells affected by diabetic conditions, and may therefore serve as sensitive early biomarkers for incipient neuronal damage caused in diabetes. Here, we comparatively assessed retinal integrity, calcium responses, and the electrophysiological profiles of specific BC types of mouse and rat organotypic retinal explants after 1 to 3 weeks in tissue culture, under moderate glucose (MG) and HG conditions. While the retinal layers of both rodent species displayed a progressively reduced thickness in culture, BCs retained their electrophysiological profiles and remained morphologically identifiable for up to 2 weeks. Responses to glutamate and endogenous inhibitory responses were routinely observed, indicating that the retinal circuitry remained intact during this period. Significant physiological differences between MG and HG conditions were evident in calcium signals and in the time course of responses to glutamate, but the voltage-gated current profiles of BCs displayed only minor variations. Overall, rat retina appeared slightly more sensitive to HG levels compared with mouse. In conclusion, electrophysiological analysis of neuronal function in rodent retinal explants is useful for the study of early damage due to HG neurotoxicity.

Spatial and temporal transcriptomic analyses reveal locust initiation of immune responses to Metarhizium acridum at the pre-penetration stage

Insect hemocyte and fat body tissues play critical functional roles in insect immunity. Little, however, is known concerning the dynamic responses of these tissues to fungal infection. Here, we report on a time course of locust hemocyte and fat body transcriptomic responses to infection by the acridid specific fungal pathogen, Metarhizium acridum. Fat body responses were more pronounced at all infection stages as compared to hemocytes. Immune and other related genes were induced far earlier than previously considered including at pre-penetration stages. Differential expression in hemocyte and fat body tissues persisted throughout the course of infection up until host death. Our data indicate selective pressure on the host to recognize the infection as early as possible in order to limit its spread. Overall, fat body and hemocyte tissues launch a robust multi-tiered response to combat the fungal pathogen, with our data providing potential host targets for exploitation in pest control.

Hair Bundle Stimulation Mode Modifies Manifestations of Mechanotransduction Adaptation.

Sound detection in auditory sensory hair cells depends on the deflection of the stereocilia hair bundle which opens mechano-electric transduction (MET) channels. Adaptation is hypothesized to be a critical property of MET that contributes to the auditory system's wide dynamic range and sharp frequency selectivity. Our recent work using a stiff probe to displace hair bundles showed that the fastest adaptation mechanism (fast adaptation) does not require calcium entry. Using fluid-jet stimuli, others obtained data showing only a calcium-dependent fast adaptation response. Because cochlear stereocilia do not move coherently and the hair cell response depends critically on the magnitude and time course of the hair bundle deflection, we developed a high-speed imaging technique to quantify this deflection in rat cochlear hair cells. The fluid jet delivers a force stimulus, and force steps lead to a complex time course of hair bundle displacement (mechanical creep), which affects the hair cell's macroscopic MET current response by masking the time course of the fast adaptation response. Modifying the fluid-jet stimulus to generate a hair bundle displacement step produced rapidly adapting currents that did not depend on membrane potential, confirming that fast adaptation does not depend on calcium entry. MET current responses differ with stimulus modality and will shape receptor potentials of different hair cell types based on their in vivo stimulus mode. These transformations will directly affect how stimuli are encoded.SIGNIFICANCE STATEMENT Mechanotransduction by sensory hair cells represents a key first step for the sound sensing ability in vertebrates. The sharp frequency tuning and wide dynamic range of sound sensation are hypothesized to require a mechanotransduction adaptation mechanism. Recent work indicated that the apparent calcium dependence of the fastest adaptation differs with the method of cochlear hair cell stimulation. Here, we reconcile existing data and show that calcium entry does not drive the fastest adaptation process, independent of the stimulation method. With force stimulation of the hair bundle, adaptation manifests differently than with displacement stimulation, indicating that the stimulation mode of the hair bundle will affect the hair cell receptor current and stimulus coding.

Time-Course Responses of Muscle-Specific MicroRNAs Following Acute Uphill or Downhill Exercise in Sprague-Dawley Rats.

Objective: The physiological characteristics and acute responses underpinning uphill running differ from those of downhill running and remain less understood. This study aimed to evaluate time-course changes of muscle-specific microRNA (miRNA) responses in striated muscle or circulation in response to uphill and downhill running.Methods: Male Sprague-Dawley rats (n = 84) were randomly assigned to a sedentary group (n = 12) and an exercise group (n = 72). The exercise group performed 90 min of uphill or downhill running. The striated muscle (quadriceps, gastrocnemius, soleus, and cardiac muscle) or circulation (plasma, exosome, exosome-free) levels of six muscle-specific miRNAs (miR-1, miR-133a, miR-133b, miR-206, miR-208a, and miR-499) were assessed at rest, immediately following exercise, and during recovery (1 h and 48 h).Results: Our results show that miR-1 and miR-133a levels are both decreased in quadriceps following downhill running (p < 0.05) while there is no change after uphill running (p > 0.05). In gastrocnemius, both uphill and downhill running decreased miR-1 level immediately after exercise and returned to baseline during recovery (p < 0.05): interestingly, only miR-499 significantly increased following uphill running (p > 0.05). Of the cell-free miRNAs in circulation, only the miR-133b levels in plasma were not affected following uphill running (p > 0.05); the other miRNA levels significantly increased immediately after exercise (p < 0.05), decreased at 1 h and significantly increased at 48 h after exercise (p < 0.05). All selected miRNA levels in exosomes were not affected following uphill running (p > 0.05), while all selected miRNA levels significantly increased during early recovery after downhill running (p > 0.05). In addition, only the miR-133a level in the exosome-free condition showed significant changes following uphill running (p < 0.05), while miR-1, miR-133a, and miR-499 levels showed significant changes after downhill running (p < 0.05).Conclusion: The results indicate that miRNA undergoes dynamic changes in tissue may play an important role in regulating different stress/adaptation following uphill and downhill running. It is likely that changed miRNA levels in plasma may act as a new biomarker for monitoring whole muscular stress during recovery.

P6308Use of cell-embedded fully implantable biosensor system for real-time, in-vivo monitoring of NF-Kappa B inflammatory pathway

Abstract Background 21st century health care remains anchored to sporadic measurements of traditional physiologic variables such as heart rate, blood pressure, weight, physical exam findings and general biochemical values. Currently there is no sensor capable of monitoring in intra-cellular protein and gene level signaling in real-time and in-vivo. This gap between biological signaling and its translation into clinically relevant therapeutics targeting the individual has limited precision medicine approaches to heart and vascular diseases. Inflammatory processes have been implicated in numerous cardiovascular diseases providing an ideal target for using Biologically based-Implantable Electronic Devices (BIED) approaches. Objective We aimed to test an implantable electro-photonic biosensor system in which living cells are integrated within the BIED and 1) serve as the primary sensor element providing in-vivo, real-time monitoring of intra-cellular processes such as gene expression, protein signaling, and target pathway activation, and 2) provide intelligent biologically based-processing in which the the output reflects biological response to an event. The engineered sensor cells provide real-time monitoring and respond to prespecified biologic signals using green fluorescence protein (GFP). The fluorescence is then detected via the BIED's photonic system and the cellular response data transmitted providing remote monitoring capabilities to facilitate the development of innovative personalized therapeutics. Methods A biologically-based implantable biosensor (BIED) platform which provides fluorescence detection, data acquisition and transmission from living cells integrated within the device was tested. The sensor cells communicate with the surrounding implant environment via a biomembrane. NRK rat cells engineered to express GFP in response to NF-κB pathway activation were cultured and housed within the sensor. Prior implant studies confirmed NRK sensor cells remained viable for 21 days in-vivo as part of a fully functional implanted BIED hardware system. Ex-vivo experiments characterized the expected magnitude and time course of NRK response to TNF-α and Lipopolysaccharide (LPS) exposure used to elicit a proinflammatory inflammatory response. The biosensor was implanted in the subcutaneous space of male Sprague Dawley rats (n=2) for a total of 11 days consisting of a baseline post-surgical recovery period of 7 days, with subsequent challenge with intraperitoneal LPS on Day 8 and 96 hours of post LPS monitoring. Results Rats implanted with the Biological based sensor and challenged with intraperitoneal LPS showed real-time expression of GFP under NF-κB transcriptional control following time course similar to ex-vivo experiments (p&lt;0.05) (Figure 1). Figure 1. Implantable Cell-Embedded Biosensor Conclusion We present the first in-vivo use of a new class of BIEDs to detect biological cell response which may herald real-time personalize health management at the molecular and cellular level. Acknowledgement/Funding Clinical and Translational Sciences Institute-University of Rochester, Efferent Labs

Immune Responses and Anti-inflammatory Strategies in a Clinically Relevant Model of Thromboembolic Ischemic Stroke with Reperfusion.

The poor clinical relevance of experimental models of stroke contributes to the translational failure between preclinical and clinical studies testing anti-inflammatory molecules for ischemic stroke. Here, we (i) describe the time course of inflammatory responses triggered by a thromboembolic model of ischemic stroke and (ii) we examine the efficacy of two clinically tested anti-inflammatory drugs: Minocycline or anti-CD49d antibodies (tested in stroke patients as Natalizumab) administered early (1h) or late (48h) after stroke onset. Radiological (lesion volume) and neurological (grip test) outcomes were evaluated at 24h and 5days after stroke. Immune cell responses peaked 48h after stroke onset. Myeloid cells (microglia/macrophages, dendritic cells, and neutrophils) were already increased 24h after stroke onset, peaked at 48h, and remained increased-although to a lesser extent-5days after stroke onset. CD8+ and CD4+ T-lymphocytes infiltrated the ipsilateral hemisphere later on (only from 48h). These responses occurred together with a progressive blood-brain barrier leakage at the lesion site, starting 24h after stroke onset. Lesion volume was maximal 24-48h after stroke onset. Minocycline reduced both lesion volume and neurological deficit only when administered early after stroke onset. The blockade of leukocyte infiltration by anti-CD49d had no impact on lesion volume or long-term neurological deficit, independently of the timing of treatment. Our data are in accordance with the results of previous clinical reports on the use of Minocycline and Natalizumab on ischemic stroke. We thus propose the use of this clinically relevant model of thromboembolic stroke with recanalization for future testing of anti-inflammatory strategies for stroke.