Effect of stress on the rehabilitation performance of rats with repetitive mild fluid percussion-induced traumatic brain injuries.

Animal models of traumatic brain injury (TBI) have shown that impaired motor and cognitive function can be improved by physical exercise. However, not each animal with TBI can be well rehabilitated at the same training intensity due to a high inter-subject variability. Hence, this paper presents a two-stage wheel-based mixed-mode rehabilitation mechanism by which the effect of stress on the rehabilitation performance was investigated. The mixed-mode rehabilitation mechanism consists of a two-week adaptive and a one-week voluntary rehabilitation program as Stages 1 and 2, respectively. In Stage 1, the common over and undertraining problem were completely resolved due to the adaptive design, and rats ran voluntarily over a 30-min duration in Stage 2. The training intensity adapted to the physical condition of all the TBI rats at all times in Stage 1, and then the self-motivated running rats were further rehabilitated under the lowest level of stress in Stage 2. For comparison purposes, another group of rats took a 3-week adaptive rehabilitation program. During the 3-week program, the rehabilitation performance of the rats were assessed using modified neurologic severity score (mNSS) and an 8-arm radial maze. Surprisingly, the group taking the mixed mode program turned out to outperform its counterpart in terms of mNSS. The mixed-mode rehabilitation mechanism was validated as an effective and efficient way to help rats restore motor, neurological and cognitive function after TBI. It was validated that the rehabilitation performance can be optimized under the lowest level of stress.

SeqCP: A sequence-based algorithm for searching circularly permuted proteins.

Circular permutation (CP) is a protein sequence rearrangement in which the amino- and carboxyl-termini of a protein can be created in different positions along the imaginary circularized sequence. Circularly permutated proteins usually exhibit conserved three-dimensional structures and functions. By comparing the structures of circular permutants (CPMs), protein research and bioengineering applications can be approached in ways that are difficult to achieve by traditional mutagenesis. Most current CP detection algorithms depend on structural information. Because there is a vast number of proteins with unknown structures, many CP pairs may remain unidentified. An efficient sequence-based CP detector will help identify more CP pairs and advance many protein studies. For instance, some hypothetical proteins may have CPMs with known functions and structures that are informative for functional annotation, but existing structure-based CP search methods cannot be applied when those hypothetical proteins lack structural information. Despite the considerable potential for applications, sequence-based CP search methods have not been well developed. We present a sequence-based method, SeqCP, which analyzes normal and duplicated sequence alignments to identify CPMs and determine candidate CP sites for proteins. SeqCP was trained by data obtained from the Circular Permutation Database and tested with nonredundant datasets from the Protein Data Bank. It shows high reliability in CP identification and achieves an AUC of 0.9. SeqCP has been implemented into a web server available at: http://pcnas.life.nthu.edu.tw/SeqCP/.

Electronic Properties of Hexagonal Graphene Quantum Rings from TAO-DFT.

The reliable prediction of electronic properties associated with graphene nanosystems can be challenging for conventional electronic structure methods, such as Kohn-Sham (KS) density functional theory (DFT), due to the presence of strong static correlation effects in these systems. To address this challenge, TAO (thermally assisted occupation) DFT has been recently proposed. In the present study, we employ TAO-DFT to predict the electronic properties of n-HGQRs (i.e., the hexagonal graphene quantum rings consisting of n aromatic rings fused together at each side). From TAO-DFT, the ground states of n-HGQRs are singlets for all the cases investigated (n = 3-15). As the system size increases, there should be a transition from the nonradical to polyradical nature of ground-state n-HGQR. The latter should be intimately related to the localization of active TAO-orbitals at the inner and outer edges of n-HGQR, which increases with increasing system size.

A robust bitmap-based real-time position tracking algorithm for rats in radial arm maze tests.

This paper aims to develop a position tracking algorithm by which a rat in a radial arm maze can be accurately located in real time. An infrared (IR) night-vision camera was hung above the maze to capture IR images of the rat. The IR images were binarized and then duplicated for subsequent intersection and opening operations. Due to simple operations and a high robustness against the noise spots formed by the droppings of the rat, it took just minutes to process more than 9000 frames, and an accuracy above 99% was reached as well. The maze was intruded by an experimenter to further test the robustness, and the accuracy slightly fell to 98%. For comparison purposes, the same experiments were carried out using a pre-trained YOLO v2 model. The YOLO counterpart gave an accuracy beyond 97% in the absence and in the presence of the intruder. In other words, this work slightly outperformed the YOLO counterpart in terms of the accuracy in both cases, which indicates the robustness of this work. However, it took the YOLO counterpart an hour or so to locate a rat contained in the frames, which highlights the contribution of this work.

Novel Stable Capacitive Electrocardiogram Measurement System.

This study presents a noncontact electrocardiogram (ECG) measurement system to replace conventional ECG electrode pads during ECG measurement. The proposed noncontact electrode design comprises a surface guard ring, the optimal input resistance, a ground guard ring, and an optimal voltage divider feedback. The surface and ground guard rings are used to reduce environmental noise. The optimal input resistor mitigates distortion caused by the input bias current, and the optimal voltage divider feedback increases the gain. Simulated gain analysis was subsequently performed to determine the most suitable parameters for the design, and the system was combined with a capacitive driven right leg circuit to reduce common-mode interference. The present study simulated actual environments in which interference is present in capacitive ECG signal measurement. Both in the case of environmental interference and motion artifact interference, relative to capacitive ECG electrodes, the proposed electrodes measured ECG signals with greater stability. In terms of R-R intervals, the measured ECG signals exhibited a 98.6% similarity to ECGs measured using contact ECG systems. The proposed noncontact ECG measurement system based on capacitive sensing is applicable for use in everyday life.

Pleiotropic mutations can rapidly evolve to directly benefit self and cooperative partner despite unfavorable conditions.

Cooperation, paying a cost to benefit others, is widespread. Cooperation can be promoted by pleiotropic 'win-win' mutations which directly benefit self (self-serving) and partner (partner-serving). Previously, we showed that partner-serving should be defined as increased benefit supply rate per intake benefit. Here, we report that win-win mutations can rapidly evolve even under conditions unfavorable for cooperation. Specifically, in a well-mixed environment we evolved engineered yeast cooperative communities where two strains exchanged costly metabolites, lysine and hypoxanthine. Among cells that consumed lysine and released hypoxanthine, ecm21 mutations repeatedly arose. ecm21 is self-serving, improving self's growth rate in limiting lysine. ecm21 is also partner-serving, increasing hypoxanthine release rate per lysine consumption and the steady state growth rate of partner and of community. ecm21 also arose in monocultures evolving in lysine-limited chemostats. Thus, even without any history of cooperation or pressure to maintain cooperation, pleiotropic win-win mutations may readily evolve to promote cooperation.

Individualized Running Wheel System with a Dynamically Adjustable Exercise Area and Speed for Rats Following Ischemic Stroke.

BACKGROUND An innovative animal running wheel with an individualized design was implemented for the rehabilitation of rats following ischemic stroke. MATERIAL AND METHODS The design of the running wheel platform included the running wheel and a side plate for exercise area adjustments. A U-curve with a width of 2 cm was drawn on the lower half of the side plate for the dynamic adjustments of five infrared (IR) sensors based on the physical fitness of the rats. The individualized training process for this running wheel consisted of 2 days of free training to record their average and maximum speeds, 3 days of progressive training to determine their exercise areas, and 2 weeks of normal training based on their average speeds, maximum speeds, and exercise areas. Blood samples were obtained from the tail veins of all rats before the operations and on Days 14, 21, and 28 postsurgery to measure cortisol levels. The motor function tests were performed on Days 7 and 28 postsurgery. On Day 28 postsurgery, the rats were sacrificed under anesthesia, and their brains were removed for Nissl and H&E staining. RESULTS On Day 28 after surgery, the motor function, lesion volume, and cell damage of the DEARW and control groups differed significantly, indicating that this device is effective for stroke rehabilitation. CONCLUSIONS The outcomes of the rats that were rehabilitated using the newly designed training system were better than those of their control-group counterparts, indicating the advantages of this designed system.

Development of a three-channel automatic climbing training system for rat rehabilitation after ischemic stroke.

This paper reports the development of a three-channel automatic speed-matching climbing training system that could train three rats at the same time for rehabilitation after an ischemic stroke. An infrared (IR) remote sensor was installed at the end of each channel to monitor the real-time position of a climbing rat. This research was carried out in five stages: i) system design; ii) hardware circuit; iii) running speed control; iv) functional testing; and v) verification using an animal model of cerebral stroke. The rehabilitated group significantly outperformed the middle cerebral artery occlusion (MCAo) sedentary group in the rota-rod and inclined plate tests 21 days after a stroke. The rehabilitated group also had a cerebral infarction volume of 28.34±19.4%, far below 56.81±18.12% of the MCAo group 28 days after the stroke, validating the effectiveness of this training platform for stroke rehabilitation. The running speed of the climbing rehabilitation training platform was designed to adapt to the physical conditions of subjects, and overtraining injuries can be completely prevented accordingly.

Noise-Resistant CECG Using Novel Capacitive Electrodes.

For years, capacitive electrocardiogram (CECG) has been known to be susceptible to ambient interference. In light of this, a novel capacitive electrode was developed as an effective way to reduce the interference effect. This was done by simply introducing the capacitive elector in series with a 1 pF capacitor, and the 60 Hz common mode noise induced by AC power lines was cancelled using a capacitive right leg (CRL) circuit. The proposed electrode did as expected outperform two counterparts in terms of SNR, and particularly gave an up to 99.8% correlation between RRIs extracted from an ECG and a CECG signal, a figure far beyond 52% and 63% using the two counterparts. This capacitive electrode was originally designed for long-term noncontact monitoring of heart rate, and hopefully can be integrated to portable devices for other medical care services in the near future.

Development of a three-channel automatic climbing training system for rat rehabilitation after ischemic stroke

This paper reports the development of a three-channel automatic speed-matching climbing training system that could train three rats at the same time for rehabilitation after an ischemic stroke. An infrared (IR) remote sensor was installed at the end of each channel to monitor the real-time position of a climbing rat. This research was carried out in five stages: i) system design; ii) hardware circuit; iii) running speed control; iv) functional testing; and v) verification using an animal model of cerebral stroke. The rehabilitated group significantly outperformed the middle cerebral artery occlusion (MCAo) sedentary group in the rota-rod and inclined plate tests 21 days after a stroke. The rehabilitated group also had a cerebral infarction volume of 28.34±19.4%, far below 56.81±18.12% of the MCAo group 28 days after the stroke, validating the effectiveness of this training platform for stroke rehabilitation. The running speed of the climbing rehabilitation training platform was designed to adapt to the physical conditions of subjects, and overtraining injuries can be completely prevented accordingly.

Remarkable cell recovery from cerebral ischemia in rats using an adaptive escalator-based rehabilitation mechanism.

Currently, many ischemic stroke patients worldwide suffer from physical and mental impairments, and thus have a low quality of life. However, although rehabilitation is acknowledged as an effective way to recover patients' health, there does not exist yet an adaptive training platform for animal tests so far. For this sake, this paper aims to develop an adaptive escalator (AE) for rehabilitation of rats with cerebral ischemia. Rats were observed to climb upward spontaneously, and a motor-driven escalator, equipped with a position detection feature and an acceleration/deceleration mechanism, was constructed accordingly as an adaptive training platform. The rehabilitation performance was subsequently rated using an incline test, a rotarod test, the infarction volume, the lesion volume, the number of MAP2 positive cells and the level of cortisol. This paper is presented in 3 parts as follows. Part 1 refers to the escalator mechanism design, part 2 describes the adaptive ladder-climbing rehabilitation mechanism, and part 3 discusses the validation of an ischemic stroke model. As it turned out, a rehabilitated group using this training platform, designated as the AE group, significantly outperformed a control counterpart in terms of a rotarod test. After the sacrifice of the rats, the AE group gave an average infarction volume of (34.36 ± 3.8)%, while the control group gave (66.41 ± 3.1)%, validating the outperformance of the escalator-based rehabilitation platform in a sense. An obvious difference between the presented training platform and conventional counterparts is the platform mechanism, and for the first time in the literature rats can be well and voluntarily rehabilitated at full capacity using an adaptive escalator. Taking into account the physical diversity among rats, the training strength provided was made adaptive as a reliable way to eliminate workout or secondary injury. Accordingly, more convincing arguments can be made using this mental stress-free training platform.

Exercise Preconditioning Attenuates Neurological Injury by Preserving Old and Newly Formed HSP72-Containing Neurons in Focal Brain Ischemia Rats.

Background: Exercise preconditioning (EP+) is a useful and important procedure for the prevention of stroke. We aimed to ascertain whether EP+ protects against ischemic brain injury by preserving heat shock protein (HSP) 72-containing neurons in ischemic brain tissues. Methods: Adult male Sprague-Dawley rats (n=240) were used to assess the contribution of HSP72-containing neurons to the neuroprotective effects of EP+ on ischemic brain injury caused by transient middle cerebral artery occlusion. Results: Significant (P<0.05) increases in the percentages of both old HSP72-containing neurons (NeuN+HSP72 double positive cells) (18~20% vs. 40~50%) and newly formed HSP72-containing neurons (BrdU+NeuN+HSP72 triple positive cells); (2~3% vs. 16~20%) after 3 weeks of exercise coincided with significant (P<0.05) reductions in brain ischemia volume (250 mm3 vs. 100 mm3), brain edema (78% vs. 74% brain water content), blood-brain barrier disruption (1.5 µg/g vs. 0.7 µg/g tissue Evans Blue dye extravasation) and neurological motor deficits (neurological severity scores of 12 vs. 6 and maximal angles of 60° vs. 20°) in brain ischemia rats. Reductions in the percentages of both old (from 40~50% to 10~12%) and newly formed (from 18~20% to 5~7%) HSP72-containing neurons by gene silencing with an intracerebral injection of pSUPER small interfering RNA showed a significant (P<0.05) reversal in the neuroprotective outcomes. Our data provide an inverse correlation between the EP+-mediated increases in both old and newly formed HSP72-containing neurons and the extent of cerebral ischemic injury. Conclusions: The percentages of both old and newly formed HSP72-containing neurons are inversely correlated with the outcomes of ischemic brain injury. Additionally, preischemic treadmill exercise improves the outcomes of ischemic brain injury by preserving both the old and newly formed HSP72-containing neurons in rats.

Disentangling strictly self-serving mutations from win-win mutations in a mutualistic microbial community.

Mutualisms can be promoted by pleiotropic win-win mutations which directly benefit self (self-serving) and partner (partner-serving). Intuitively, partner-serving phenotype could be quantified as an individual's benefit supply rate to partners. Here, we demonstrate the inadequacy of this thinking, and propose an alternative. Specifically, we evolved well-mixed mutualistic communities where two engineered yeast strains exchanged essential metabolites lysine and hypoxanthine. Among cells that consumed lysine and released hypoxanthine, a chromosome duplication mutation seemed win-win: it improved cell's affinity for lysine (self-serving), and increased hypoxanthine release rate per cell (partner-serving). However, increased release rate was due to increased cell size accompanied by increased lysine utilization per birth. Consequently, total hypoxanthine release rate per lysine utilization (defined as 'exchange ratio') remained unchanged. Indeed, this mutation did not increase the steady state growth rate of partner, and is thus solely self-serving during long-term growth. By extension, reduced benefit production rate by an individual may not imply cheating.

An adaptive fall-free rehabilitation mechanism for ischemic stroke rat patients.

Today's commercial forced exercise platforms had been validated not as a well-designed rehabilitation environment for rats with a stroke, for the reason that rat with a stroke cannot take exercise at a constant intensity for a long period of time. In light of this, this work presented an adaptive, fall-free ischemic stroke rehabilitation mechanism in an animal model, which was implemented in an infrared-sensing adaptive feedback control running wheel (IAFCRW) platform. Consequently, rats with a stroke can be safely rehabilitated all the time, and particularly at full capacity for approximately one third of a training duration, in a completely fall-free environment according to individual physical differences by repeated use of an acceleration/deceleration mechanism. The performance of this platform was assessed using an animal ischemic stroke model. The IAFCRW therapy regimen was validated to outperform a treadmill and a conventional running wheel counterpart with respect to the reduction in the neurobehavioral deficits caused by middle cerebral artery occlusion (MCAo). IAFCRW is the first adaptive forced exercise training platform short of electrical stimulation-assistance in the literature, and ischemic stroke rats benefit more in terms of the behavioral tests run at the end of a 3-week rehabilitation program after a stroke thereby.

Exercise Rehabilitation Attenuates Cognitive Deficits in Rats with Traumatic Brain Injury by Stimulating the Cerebral HSP20/BDNF/TrkB Signalling Axis.

Physical exercise (PE) is an effective method for improving cognitive function among patients with traumatic brain injury (TBI). We previously demonstrated that PE with an infrared-sensing running wheel (ISRW) system provides strong neuroprotection in an experimental animal model of stroke. In this study, we used fluid percussion injury in rats to simulate mild TBI. For rats, we used both passive avoidance learning and the Y-maze tests to evaluate cognitive function. We investigated whether PE rehabilitation attenuated cognitive deficits in rats with TBI and determined the contribution of hippocampal and cortical expression of heat shock protein 20 (HSP20) to PE-mediated cognitive recovery. In addition to increasing hippocampal and cortical expression of HSP20, brain-derived neurotrophic factor (BDNF), and the tropomyosin receptor kinase B (TrkB) ratio, PE rehabilitation significantly attenuated brain contusion and improved cognitive deficits in the rat model. Furthermore, reducing hippocampal and cortical expression of HSP20 with an intracerebral injection of pSUPER hsp20 small interfering RNA significantly diminished the PE-induced overexpression of hippocampal and cortical BDNF and the TrkB ratio and also reversed the beneficial effect of PE in reducing neurotrauma and the cognitive deficits. A positive Pearson correlation was found between HSP20 and BDNF, as well as between HSP20 and TrkB, in the hippocampal and cortical tissues. We thus conclude that post-ischaemic ISRW exercise rehabilitation attenuates cognitive deficits, as well as brain contusions, in TBI rats by stimulating the cerebral HSP20/BDNF/TrkB signalling axis.

An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance.

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity for reducing the severity of the effects of the stroke in rats. This system provides more effective animal exercise training than commercially available systems such as treadmills and motorized running wheels (MRWs). In contrast to an MRW that can only achieve speeds below 20 m/min, rats are permitted to run at a stable speed of 30 m/min on a more spacious and high-density rubber running track supported by a 15 cm wide acrylic wheel with a diameter of 55 cm in this work. Using a predefined adaptive acceleration curve, the system not only reduces the operator error but also trains the rats to run persistently until a specified intensity is reached. As a way to evaluate the exercise effectiveness, real-time position of a rat is detected by four pairs of infrared sensors deployed on the running wheel. Once an adaptive acceleration curve is initiated using a microcontroller, the data obtained by the infrared sensors are automatically recorded and analyzed in a computer. For comparison purposes, 3 week training is conducted on rats using a treadmill, an MRW and a PRW. After surgically inducing middle cerebral artery occlusion (MCAo), modified neurological severity scores (mNSS) and an inclined plane test were conducted to assess the neurological damages to the rats. PRW is experimentally validated as the most effective among such animal mobility systems. Furthermore, an exercise effectiveness measure, based on rat position analysis, showed that there is a high negative correlation between the effective exercise and the infarct volume, and can be employed to quantify a rat training in any type of brain damage reduction experiments.

Antibiotic treatment enhances the genome-wide mutation rate of target cells.

Although it is well known that microbial populations can respond adaptively to challenges from antibiotics, empirical difficulties in distinguishing the roles of de novo mutation and natural selection have left several issues unresolved. Here, we explore the mutational properties of Escherichia coli exposed to long-term sublethal levels of the antibiotic norfloxacin, using a mutation accumulation design combined with whole-genome sequencing of replicate lines. The genome-wide mutation rate significantly increases with norfloxacin concentration. This response is associated with enhanced expression of error-prone DNA polymerases and may also involve indirect effects of norfloxacin on DNA mismatch and oxidative-damage repair. Moreover, we find that acquisition of antibiotic resistance can be enhanced solely by accelerated mutagenesis, i.e., without direct involvement of selection. Our results suggest that antibiotics may generally enhance the mutation rates of target cells, thereby accelerating the rate of adaptation not only to the antibiotic itself but to additional challenges faced by invasive pathogens.

Improved infrared-sensing running wheel systems with an effective exercise activity indicator.

This paper describes an infrared-sensing running wheel (ISRW) system for the quantitative measurement of effective exercise activity in rats. The ISRW system provides superior exercise training compared with commercially available traditional animal running platforms. Four infrared (IR) light-emitting diode/detector pairs embedded around the rim of the wheel detect the rat's real-time position; the acrylic wheel has a diameter of 55 cm and a thickness of 15 cm, that is, it is larger and thicker than traditional exercise wheels, and it is equipped with a rubber track. The acrylic wheel hangs virtually frictionless, and a DC motor with an axially mounted rubber wheel, which has a diameter of 10 cm, drives the acrylic wheel from the outer edge. The system can automatically train rats to run persistently. The proposed system can determine effective exercise activity (EEA), with the IR sensors (which are connected to a conventional PC) recording the rat exercise behavior. A prototype of the system was verified by a hospital research group performing ischemic stroke experiments on rats by considering middle cerebral artery occlusion. The experimental data demonstrated that the proposed system provides greater neuroprotection in an animal stroke model compared with a conventional treadmill and a motorized running wheel for a given exercise intensity. The quantitative exercise effectiveness indicator showed a 92% correlation between an increase in the EEA and a decrease in the infarct volume. This indicator can be used as a noninvasive and objective reference in clinical animal exercise experiments.

Design of a novel flexible capacitive sensing mattress for monitoring sleeping respiratory.

In this paper, an algorithm to extract respiration signals using a flexible projected capacitive sensing mattress (FPCSM) designed for personal health assessment is proposed. Unlike the interfaces of conventional measurement systems for poly-somnography (PSG) and other alternative contemporary systems, the proposed FPCSM uses projected capacitive sensing capability that is not worn or attached to the body. The FPCSM is composed of a multi-electrode sensor array that can not only observe gestures and motion behaviors, but also enables the FPCSM to function as a respiration monitor during sleep using the proposed approach. To improve long-term monitoring when body movement is possible, the FPCSM enables the selection of data from the sensing array, and the FPCSM methodology selects the electrodes with the optimal signals after the application of a channel reduction algorithm that counts the reversals in the capacitive sensing signals as a quality indicator. The simple algorithm is implemented in the time domain. The FPCSM system is used in experimental tests and is simultaneously compared with a commercial PSG system for verification. Multiple synchronous measurements are performed from different locations of body contact, and parallel data sets are collected. The experimental comparison yields a correlation coefficient of 0.88 between FPCSM and PSG, demonstrating the feasibility of the system design.

Accumulated mental stress study using the meridians of traditional Chinese medicine with photoplethysmography.

OBJECTIVES: To investigate accumulated mental stress according to the concept of the meridians of Traditional Chinese Medicine (TCM). This stress was quantified by using pulse spectrum analysis of finger-tip photoplethysmography (PPG). Stress accumulation is one of the main causes of cardiovascular disease and depression in humans, resulting in chronic physiologic malfunctions; however, few studies have thoroughly assessed the quantitative evaluation of accumulative stress using the concept of TCM. DESIGN: This study investigated accumulated mental stress from the perspective of TCM based on an 8-day experiment. The theory of organ resonance was integrated into the proposed PPG sensing instrument to capture the nine harmonics of TCM. Participants were given daily mental arithmetic tasks over 1 week to simulate stress accumulation, and trends in the proportion of the nine harmonics of TCM were extracted over several days and analyzed to identify the affective factors related to cumulative stress. RESULTS: The experimental results showed that the kidney harmonic proportion (C2) and stomach harmonic proportion (C5) were significant only on the first few days because of a physiologic phenomenon of temporary stimulation. Most important, the trend of the liver harmonic proportion (C1) from days 3 to 8 dramatically increased and became gradually saturated because of the influence of accumulated mental stress. CONCLUSIONS: The results strongly suggest that pulse spectrum analysis of the PPG signal provides physiologically and pathologically important information on accumulated mental stress and can be useful for TCM analysis.