Sensors for Caspase Activities

Cysteine proteases, caspases, play an important role in the process of programmed cell death. In apoptosis, the cell signaling pathways overlap at executioner caspases regardless of the initial stimulus. Caspase-3 is the key one. This review considers the sensors for activity of caspase-3 and some other caspases. Over the past decade, many such sensors utilizing a variety of detection principles have been created; fluorescence and luminescence are the most common detection methods. The intracellular sensors are of particular interest; they allow visualization of the activation of caspases in living systems. Herein, we briefly describe sensors of various designs, the advantages and disadvantages of which must be taken into account when choosing a sensor for a particular experimental system.

     

Predicting Recovery of Voluntary Upper Extremity Movement in Subacute Stroke Patients with Severe Upper Extremity Paresis

DesignProspective cohort study.Methods140 (out of 590) stroke patients with severe UE paresis completed all assessments. Voluntary UE movement was assessed using the UE subscale of the Stroke Rehabilitation Assessment of Movement (STREAM-UE). Two outcome measures, STREAM-UE scores at discharge (DC_STREAM-UE) and changes between admission and discharge (Δ_STREAM-UE), were investigated to represent the final states and improvement of the recovery of voluntary UE movement. Stepwise regression analyses were used to investigate 19 clinical variables and to find the best predictive models of the two outcome measures.ResultsThe participants showed wide variation in both DC_STREAM-UE and Δ_STREAM-UE. 3.6% of the participants almost fully recovered at discharge (DC_STREAM-UE > 15). A large improvement (Δ_STREAM-UE >= 10) occurred in 16.4% of the participants, while 32.9% of the participants did not have any improvement. The four predictors for the DC_STREAM-UE (R² = 35.0%) were ‘baseline STREAM-UE score’, ‘hemorrhagic stroke’, ‘baseline National Institutes of Health Stroke Scale (NIHSS) score’, and ‘cortical lesion excluding primary motor cortex’. The three predictors for the Δ_STREAM-UE (R² = 22.0%) were ‘hemorrhagic stroke’, ‘baseline NIHSS score’, and ‘cortical lesion excluding primary motor cortex’.ConclusionsRecovery of voluntary UE movement varied widely in patients with severe UE paresis after stroke. The predictive power of clinical variables was poor. Both results indicate the complex nature of voluntary UE movement recovery in patients with severe UE paresis after stroke.     

Continuous Monitoring of Body Temperature in Pigs Using Non-Invasive Ear Canal Sensors

Monitoring of the body temperature in animals kept under intensive production conditions is desirable for a number of reasons: oestrus detection, disease monitoring and stress indication in addition to more specific needs, such as the selection of breeding pigs resistant to malignant hyperthermia and for various research purposes.     

Primary Venous Obstruction of the Upper Extremity

Three cases of primary axillary and subclavian venous obstruction presented herein demonstrated the classic findings of swelling, aching or pain, and discoloration following effort in an otherwise healthy person. The clinical diagnosis was confirmed by venography. Residual changes were frequent after treatment that included anticoagulation and thrombectomy.     

Aging Contributes to Inflammation in Upper Extremity Tendons and Declines in Forelimb Agility in a Rat Model of Upper Extremity Overuse

We sought to determine if tendon inflammatory and histopathological responses increase in aged rats compared to young rats performing a voluntary upper extremity repetitive task, and if these changes are associated with motor declines. Ninety-six female Sprague-Dawley rats were used in the rat model of upper extremity overuse: 67 aged and 29 young adult rats. After a training period of 4 weeks, task rats performed a voluntary high repetition low force (HRLF) handle-pulling task for 2 hrs/day, 3 days/wk for up to 12 weeks. Upper extremity motor function was assessed, as were inflammatory and histomorphological changes in flexor digitorum and supraspinatus tendons. The percentage of successful reaches improved in young adult HRLF rats, but not in aged HRLF rats. Forelimb agility decreased transiently in young adult HRLF rats, but persistently in aged HRLF rats. HRLF task performance for 12 weeks lead to increased IL-1beta and IL-6 in flexor digitorum tendons of aged HRLF rats, compared to aged normal control (NC) as well as young adult HRLF rats. In contrast, TNF-alpha increased more in flexor digitorum tendons of young adult 12-week HRLF rats than in aged HRLF rats. Vascularity and collagen fibril organization were not affected by task performance in flexor digitorum tendons of either age group, although cellularity increased in both. By week 12 of HRLF task performance, vascularity and cellularity increased in the supraspinatus tendons of only aged rats. The increased cellularity was due to increased macrophages and connective tissue growth factor (CTGF)-immunoreactive fibroblasts in the peritendon. In conclusion, aged rat tendons were overall more affected by the HRLF task than young adult tendons, particularly supraspinatus tendons. Greater inflammatory changes in aged HRLF rat tendons were observed, increases associated temporally with decreased forelimb agility and lack of improvement in task success.     

Automated Assessment of Upper Extremity Movement Impairment due to Stroke

Current diagnosis and treatment of movement impairment post-stroke is based on the subjective assessment of select movements by a trained clinical specialist. However, modern low-cost motion capture technology allows for the development of automated quantitative assessment of motor impairment. Such outcome measures are crucial for advancing post-stroke treatment methods. We sought to develop an automated method of measuring the quality of movement in clinically-relevant terms from low-cost motion capture. Unconstrained movements of upper extremity were performed by people with chronic hemiparesis and recorded by standard and low-cost motion capture systems. Quantitative scores derived from motion capture were compared to qualitative clinical scores produced by trained human raters. A strong linear relationship was found between qualitative scores and quantitative scores derived from both standard and low-cost motion capture. Performance of the automated scoring algorithm was matched by averaged qualitative scores of three human raters. We conclude that low-cost motion capture combined with an automated scoring algorithm is a feasible method to assess objectively upper-arm impairment post stroke. The application of this technology may not only reduce the cost of assessment of post-stroke movement impairment, but also promote the acceptance of objective impairment measures into routine medical practice.     

An Accelerometry-Based Methodology for Assessment of Real-World Bilateral Upper Extremity Activity

Background

The use of both upper extremities (UE) is necessary for the completion of many everyday tasks. Few clinical assessments measure the abilities of the UEs to work together; rather, they assess unilateral function and compare it between affected and unaffected UEs. Furthermore, clinical assessments are unable to measure function that occurs in the real-world, outside the clinic. This study examines the validity of an innovative approach to assess real-world bilateral UE activity using accelerometry.

Methods

Seventy-four neurologically intact adults completed ten tasks (donning/doffing shoes, grooming, stacking boxes, cutting playdough, folding towels, writing, unilateral sorting, bilateral sorting, unilateral typing, and bilateral typing) while wearing accelerometers on both wrists. Two variables, the Bilateral Magnitude and Magnitude Ratio, were derived from accelerometry data to distinguish between high- and low-intensity tasks, and between bilateral and unilateral tasks. Estimated energy expenditure and time spent in simultaneous UE activity for each task were also calculated.

Results

The Bilateral Magnitude distinguished between high- and low-intensity tasks, and the Magnitude Ratio distinguished between unilateral and bilateral UE tasks. The Bilateral Magnitude was strongly correlated with estimated energy expenditure (ρ = 0.74, p<0.02), and the Magnitude Ratio was strongly correlated with time spent in simultaneous UE activity (ρ = 0.93, p<0.01) across tasks.

Conclusions

These results demonstrate face validity and construct validity of this methodology to quantify bilateral UE activity during the performance of everyday tasks performed in a laboratory setting, and can now be used to assess bilateral UE activity in real-world environments.     

Using an Insect Mushroom Body Circuit to Encode Route Memory in Complex Natural Environments

Ants, like many other animals, use visual memory to follow extended routes through complex environments, but it is unknown how their small brains implement this capability. The mushroom body neuropils have been identified as a crucial memory circuit in the insect brain, but their function has mostly been explored for simple olfactory association tasks. We show that a spiking neural model of this circuit originally developed to describe fruitfly (Drosophila melanogaster) olfactory association, can also account for the ability of desert ants (Cataglyphis velox) to rapidly learn visual routes through complex natural environments. We further demonstrate that abstracting the key computational principles of this circuit, which include one-shot learning of sparse codes, enables the theoretical storage capacity of the ant mushroom body to be estimated at hundreds of independent images.     

Hormonal and Neuromuscular Responses to Mechanical Vibration Applied to Upper Extremity Muscles

Objective

To investigate the acute residual hormonal and neuromuscular responses exhibited following a single session of mechanical vibration applied to the upper extremities among different acceleration loads.

Methods

Thirty male students were randomly assigned to a high vibration group (HVG), a low vibration group (LVG), or a control group (CG). A randomized double-blind, controlled-parallel study design was employed. The measurements and interventions were performed at the Laboratory of Biomechanics of the University of L''Aquila. The HVG and LVG participants were exposed to a series of 20 trials ×10 s of synchronous whole-body vibration (WBV) with a 10-s pause between each trial and a 4-min pause after the first 10 trials. The CG participants assumed an isometric push-up position without WBV. The outcome measures were growth hormone (GH), testosterone, maximal voluntary isometric contraction during bench-press, maximal voluntary isometric contraction during handgrip, and electromyography root-mean-square (EMG_rms) muscle activity (pectoralis major [PM], triceps brachii [TB], anterior deltoid [DE], and flexor carpi radialis [FCR]).

Results

The GH increased significantly over time only in the HVG (P = 0.003). Additionally, the testosterone levels changed significantly over time in the LVG (P = 0.011) and the HVG (P = 0.001). MVC during bench press decreased significantly in the LVG (P = 0.001) and the HVG (P = 0.002). In the HVG, the EMG_rms decreased significantly in the TB (P = 0.006) muscle. In the LVG, the EMG_rms decreased significantly in the DE (P = 0.009) and FCR (P = 0.006) muscles.

Conclusion

Synchronous WBV acutely increased GH and testosterone serum concentrations and decreased the MVC and their respective maximal EMG_rms activities, which indicated a possible central fatigue effect. Interestingly, only the GH response was dependent on the acceleration with respect to the subjects'' responsiveness.     

Glutamine Flux Imaging Using Genetically Encoded Sensors

Genetically encoded sensors allow real-time monitoring of biological molecules at a subcellular resolution. A tremendous variety of such sensors for biological molecules became available in the past 15 years, some of which became indispensable tools that are used routinely in many laboratories.One of the exciting applications of genetically encoded sensors is the use of these sensors in investigating cellular transport processes. Properties of transporters such as kinetics and substrate specificities can be investigated at a cellular level, providing possibilities for cell-type specific analyses of transport activities. In this article, we will demonstrate how transporter dynamics can be observed using genetically encoded glutamine sensor as an example. Experimental design, technical details of the experimental settings, and considerations for post-experimental analyses will be discussed.     

肌电生物反馈对急性期脑卒中偏瘫患者上肢功能的影响

目的:评价肌电生物反馈联合常规康复训练对急性期脑卒中偏瘫患者上肢功能的影响,为脑卒中后急性期上肢运动功能障碍的康复治疗探寻更有效的方法.方法:将35例生命体征平稳、无意识障碍的急性期脑卒中偏瘫患者分为观察组18例和对照组17例.两组患者均给予神经内科药物治疗和常规康复训练,观察组患者偏瘫侧上肢加以肌电生物反馈治疗.两组均于治疗前和治疗2个月后对偏瘫侧上肢进行Bmnnstrom运动功能分级、简式Fugl-Meyer(FMA)上肢运动功能评定和腕背伸时肌肉最大收缩时肌电(EMG)幅值的测定.结果:经2个月治疗后,两组患者偏瘫侧上肢的Bnnnstrom分级、FMA及EMG幅值较治疗前均有提高(P＜0.05),且观察组各项评分均优于对照组(P＜0.05).结论:本文中观察组和对照组的患者经康复治疗后,偏瘫侧上肢的运动功能均较治疗前提高.但在脑卒中患者的急性期应用肌电生物反馈治疗联合常规康复训练,较单独使用常规运动疗法和作业疗法治疗,能更好地改善急性期脑卒中偏瘫患者上肢的功能,更显著地促进偏瘫上肢的运动功能恢复及分离运动的产生,同时对促进腕关节背伸肌力的恢复方面提供了电生理学依据.总之,在常规康复训练的基础上加用肌电生物反馈治疗,可更显著地促进急性期脑卒中偏瘫患者的上肢功能,为脑卒中急性期肢体运动障碍的康复治疗提供了更有效的方法.     

A Keratin Antibody Recognizing a Heterotypic Complex: Epitope Mapping to Complementary Locations on Both Components of the Complex

Keratin filaments in simple epithelial cells are heteropolymers of keratin 8 (K8) and keratin 18 (K18), which can be stained by the monoclonal antibody (MAb) LE61. This antibody has been widely used to study keratin expression in normal and neoplastic tissues. In this study we have found that MAb LE61 does not react with individual keratin polypeptides either derived from natural sources or expressed as recombinant proteins inEscherichia coli.However, when K8 or K18 bound to nitrocellulose were incubated with complementary keratin they became reactive with this antibody. A mixture of K8 and K18 in solution also reacted strongly with the MAb LE61 in ELISA. These observations suggest that the antibody recognizes a discontinuous epitope on the keratin complex. The antibody also reacted with complexes of K8 and K18 with other keratins. To locate the epitope of this antibody we have expressed K8 and K18 fragments, deleted from the amino- and carboxyl-termini, as fusion proteins with glutathioneS-transferase. These fragments were able to form a heterotypic complex with the complementary keratin. Binding of the MAb LE61 to these complexes mapped the two halves of the epitope on K8, between residues 353 and 367, and on K18, between residues 357 and 385. The two halves of the epitope appear to be in close association in the heterotypic complex since deletions from the amino-terminus did not influence the antibody binding. The highly conserved nature of this epitope in both type I and type II keratins could explain the MAb LE61 reactivity with complexes of K8 or K18 with other keratins.     

Intersegmental Eye-Head-Body Interactions during Complex Whole Body Movements

Using state-of-the-art technology, interactions of eye, head and intersegmental body movements were analyzed for the first time during multiple twisting somersaults of high-level gymnasts. With this aim, we used a unique combination of a 16-channel infrared kinemetric system; a three-dimensional video kinemetric system; wireless electromyography; and a specialized wireless sport-video-oculography system, which was able to capture and calculate precise oculomotor data under conditions of rapid multiaxial acceleration. All data were synchronized and integrated in a multimodal software tool for three-dimensional analysis. During specific phases of the recorded movements, a previously unknown eye-head-body interaction was observed. The phenomenon was marked by a prolonged and complete suppression of gaze-stabilizing eye movements, in favor of a tight coupling with the head, spine and joint movements of the gymnasts. Potential reasons for these observations are discussed with regard to earlier findings and integrated within a functional model.     

Using Friends as Sensors to Detect Global-Scale Contagious Outbreaks

Recent research has focused on the monitoring of global–scale online data for improved detection of epidemics, mood patterns, movements in the stock market political revolutions, box-office revenues, consumer behaviour and many other important phenomena. However, privacy considerations and the sheer scale of data available online are quickly making global monitoring infeasible, and existing methods do not take full advantage of local network structure to identify key nodes for monitoring. Here, we develop a model of the contagious spread of information in a global-scale, publicly-articulated social network and show that a simple method can yield not just early detection, but advance warning of contagious outbreaks. In this method, we randomly choose a small fraction of nodes in the network and then we randomly choose a friend of each node to include in a group for local monitoring. Using six months of data from most of the full Twittersphere, we show that this friend group is more central in the network and it helps us to detect viral outbreaks of the use of novel hashtags about 7 days earlier than we could with an equal-sized randomly chosen group. Moreover, the method actually works better than expected due to network structure alone because highly central actors are both more active and exhibit increased diversity in the information they transmit to others. These results suggest that local monitoring is not just more efficient, but also more effective, and it may be applied to monitor contagious processes in global–scale networks.