CE Run Research Articles

Lifelong Outcomes of Systemic Adeno-Associated Virus Micro-Dystrophin Gene Therapy in a Murine Duchenne Muscular Dystrophy Model.

Adeno-associated virus (AAV)-mediated systemic micro-dystrophin (μDys) therapy is currently in clinical trials. The hope is to permanently improve the life quality of Duchenne muscular dystrophy (DMD) patients. Numerous preclinical studies have been conducted to support these trials. However, none examined whether a single therapy at a young age can lead to lifelong disease amelioration. To address this critical question, we injected 1 × 1013 vg particles/mouse of an AAV serotype-9 μDys vector to 3-month-old mdx mice through the tail vein. Therapeutic outcomes were evaluated at the age of 11 months (adulthood, 8 months postinjection) and 21 months (terminal age, 18 months postinjection). Immunostaining and Western blot showed saturated supraphysiological levels of μDys expression in skeletal muscle and heart till the end of the study. Treatment significantly improved grip force and treadmill running, and significantly reduced the serum creatine kinase level at both time points. Since cardiac death is a major threat in late-stage patients, we evaluated cardiac electrophysiology and hemodynamics by ECG and the closed-chest cardiac catheter assay, respectively. Significant improvements were observed in these assays. Importantly, many ECG and hemodynamic parameters (heart rate, PR interval, QRS duration, QTc interval, end-diastolic/systolic volume, dP/dt max and min, max pressure, and ejection fraction) were completely normalized at 21 months of age. Our results have provided direct evidence that a single systemic AAV μDys therapy has the potential to provide lifelong benefits in the murine DMD model.

Robotic friction stir welding – seam-tracking control, force control and process supervision

Purpose This study aims to enable robotic friction stir welding (FSW) in practice. The use of robots has hitherto been limited, because of the large contact forces necessary for FSW. These forces are detrimental for the position accuracy of the robot. In this context, it is not sufficient to rely on the robot’s internal sensors for positioning. This paper describes and evaluates a new method for overcoming this issue. Design/methodology/approach A closed-loop robot control system for seam-tracking control and force control, running and recording data in real-time operation, was developed. The complete system was experimentally verified. External position measurements were obtained from a laser seam tracker and deviations from the seam were compensated for, using feedback of the measurements to a position controller. Findings The proposed system was shown to be working well in overcoming position error. The system is flexible and reconfigurable for batch and short production runs. The welds were free of defects and had beneficial mechanical properties. Research limitations/implications In the experiments, the laser seam tracker was used both for control feedback and for performance evaluation. For evaluation, it would be better to use yet another external sensor for position measurements, providing ground truth. Practical implications These results imply that robotic FSW is practically realizable, with the accuracy requirements fulfilled. Originality/value The method proposed in this research yields very accurate seam tracking as compared to previous research. This accuracy, in turn, is crucial for the quality of the resulting material.

Quadriceps strength symmetry predicts vertical ground reaction force symmetry during running in patients who have undergone ACL reconstruction

ObjectiveTo determine whether quadriceps strength symmetry can predict peak vertical ground reaction force (vGRF) running force symmetry in patients who have undergone ACL reconstruction (ACLR). We also sought to determine a cutoff for quadriceps strength symmetry to identify patients at risk for vGRF running asymmetry. DesignRetrospective cross-sectional. SettingClinical facility. MethodsBilateral quadriceps strength and vGRF data during running were obtained from 79 patients 26–30 weeks post ACLR. Linear regression was used to determine if quadriceps strength symmetry predicted peak vGRF running force symmetry. Classification and regression tree (CART) analysis was used to determine the cutoff value for quadriceps strength symmetry to identify patients at risk for vGRF running asymmetry. ResultsIncreased quadriceps strength symmetry predicted increased vGRF running symmetry (R2 = 0.20). CART analysis revealed that patients with quadriceps strength symmetry less than or equal to 88% were at highest risk for vGRF running asymmetry (R2 = 26%). ConclusionGreater quadriceps strength symmetry is predictive of greater vGRF running force symmetry in patients who have undergone ACLR. This finding highlights the need for clinicians to consider the degree of quadriceps strength symmetry before initiating a return to running program.

Research on Force Characteristics and Running Performance of Novel Type High-Temperature Superconductor Magnetic Levitation Vehicle

Research on Force Characteristics and Running Performance of Novel Type High-Temperature Superconductor Magnetic Levitation Vehicle

An online target and rapid screening method for α-glucosidase inhibitors based on capillary electrophoresis.

Screening enzymatic active compounds is one of the important fields in drug research. α-Glucosidase can hydrolyze carbohydrates to monosaccharides after meals and lead to the rise of blood glucose levels in human body. Thus, the inhibition of α-glucosidase activity is an effective approach for the diabetes treatment. In this work, we developed a new method to simultaneously screen multiple bioactive compounds within a single CE running. The affect factors on the method performance, including injection, mixing, incubation, separation and detection, were carefully analyzed and discussed. Under the optimum, the mixture consisting of two internal standards (DMSO and 4-nitrophenol) and five compounds (lyoniresinol, hydroxytyrosol, rutin, kaempferol, and quercetin) was simultaneously screened, and kaempferol and quercetin showed stronger activity and this conclusion was also supported by offline assay. Furthermore, molecular docking was employed for investigating its interaction mechanism. Eventually, the established method has been applied to screen potential α-glucosidase inhibitors from an extract of Lycium barbarum and the peak area of rutin, taxifolin, quercetin, and chlorogenic acid in L. barbarum samples changed before and after the enzymatic reaction, confirming that these four compounds had potential inhibitory activities, which was consistent with the literature data. The present work provides a promising method for the target and rapid discovery of bioactive compounds from a plant extract or mixture.

Comparison of murine behavioural and physiological responses after forced exercise by electrical shock versus manual prodding.

What is the central question of this study? Forced treadmill exercise using electrical shock is the most common technique in rodent exercise studies. Here, we examined how the use of electrical shock during forced treadmill exercise affects behavioural and physiological responses in comparison to a novel non-electrical shock technique. What is the main finding and its importance? In comparison to mice that underwent traditional treadmill running induced by electrical shock, mice that underwent forced running using a novel technique involving gentle prodding to induce running showed: (i) higher locomotor activity; (ii) less anxiety-like behaviour; and (iii) altered exercise-induced muscle pain immediately after exercise. Animal models of exercise have been useful to understand underlying cellular and molecular mechanisms. Many studies have used methods of exercise that are unduly stressful (e.g., electrical shock to force running), potentially skewing results. Here, we compared physiological and behavioural responses of mice after exercise induced using a prodding technique that avoids electrical shock versus a traditional protocol using electrical shock. We found that exercise performance was similar for both techniques; however, the shock group demonstrated significantly lower locomotor activity and higher anxiety-like behaviour. We also observed divergent effects on muscle pain; the prodding group showed hyperalgesia immediately after exercise, whereas the shock group showed hypoalgesia. Corticosterone concentrations were elevated to a similar extent for both groups. In conclusion, mice that were exercised without shock generated similar maximal exercise performance, but postexercise these mice showed an increase in locomotor activity, less anxiety-like behaviour and altered muscle pain in comparison to mice that exercised with shock. Our data suggest that running of mice without the use of electrical shock is potentially less stressful and might be a better technique to study the physiological and behavioural responses to exercise.

ASSISTED EMPLOYMENT AND SELF-EMLOPYMENT FOR THE PEOPLE WITH DISABILITIES

He aim of this study is to present legal regulations concerning professional activation of disabled people. In the Polish legal system there are various forms of support for professional activation of people with disabilities, starting from taking up employment by a disabled person in a sheltered workplace, hospital, university, offi ce or company run in the form of a sole proprietorship. Professional activation and reduction of unemployment among disabled people remains a challenge. People with disabilities are often isolated in the labour market, which contributes to the deterioration of their living conditions. The problem of professional activation can and should be considered from two perspectives: employment and self-employment. The paper presents various types of instruments that can be used in the process of professional activation of people with disabilities.

The relationship between foot arch flexibility and medial-lateral ground reaction force distribution

BackgroundOveruse running injury susceptibility has previously been associated with the magnitude and slope of ground reaction force profiles, most often in the vertical axis. However, despite the implications of excessive pronation and supination on injury susceptibility, very little research has examined the factors that might affect distribution of force in the medial-lateral directions. Research questionThe purpose of this study was to consider how foot structure, specifically arch flexibility, affects the distribution of ground reaction force between the medial-lateral and vertical planes of motion. MethodsTwenty-five participants were classified as having stiff or flexible arches, and three dimensional kinetic data were gathered while the volunteers ran at 7 mph on an instrumented treadmill. A mixed-effects ANOVA was used to analyze the effect of arch flexibility type on distribution of ground reaction force impulse in the medial and lateral directions. ResultsThe results suggest that individuals with relatively stiff arches experience a greater proportion of ground reaction force in the medial-lateral plane of motion, as compared with those with more flexible arches (p = 0.03). Further, the results suggest that most individuals, regardless of foot structure, experience greater impulse of force in the lateral than in the medial direction (p < 0.01). SignificanceConsidering previously explored relationships between ground reaction force, foot pronation/supination, and chronic running injuries, the results of this study suggest that arch flexibility could be used as a criterion for assessing injury susceptibility. Further, conclusions drawn from this study add to the discussion on the pros and cons of training or using devices to increase or restrict arch flexibility while running.

Effects of a 24 Week Multifaceted Sports Training Program on Some Physical Characteristics of 5 to 9 Year-Old Children

This study was conducted to investigate the effects of a 24-week multifaceted sport training program on some physical and performance characteristics of 5-9 year old children. There are many researches about the necessities for children to start physical activity at an early age. According to the characteristics of different physical activities to be performed, children can endow with different developments. Multifaceted development in the motor properties of an unbranched individual will also benefit him/her in the branching phase. A branch that will be applied to children will develop the characteristics of that branch in the child. This study aims to determine the effects of the implementation of a training program consisting of the basic characteristics of different branches on the physical characteristics of children.A group of children with a mean age of 6.4 (SD 1.34) (n = 98) residing in Ankara participated in the study. A training program including basic techniques of many sports branches was applied to children in the form of 2 lesson hours per day for 2 days in each week. Lessons are scheduled for 45 minutes. During the 24-week training period, children were tested and measured three times; at the beginning, in the middle and at the end of the study. All physical characteristics of the children within the scope of the study has improved, and the differences in the test of vertical jump, throwing medicine ball, right foot balance posture, left hand grip force, shuttle, horizontal jump and 10x5m running were found to be statistically significant (p&lt;0.05).

A stochastic model for the integrated optimization on metro timetable and speed profile with uncertain train mass

The integrated timetable and speed profile optimization model has recently attracted more attention because of its good achievements on energy conservation in metro systems. However, most previous studies often ignore the spatial and temporal uncertainties of train mass, and the variabilities of tractive force, braking force and basic running resistance on energy consumption in order to simplify the model formulation and solution algorithm. In this paper, we develop an integrated metro timetable and speed profile optimization model to minimize the total tractive energy consumption, where these real-world operating conditions are explicitly considered in the model formulation and solution algorithm. Firstly, we formulate a two-phase stochastic programming model to determine the timetable and speed profile. Given the speed profile, the first phase determines the timetable by scheduling the arrival and departure times for each station, and the second phase determines the speed profile for each inter-station with the scheduled arrival and departure times. Secondly, we design a simulation-based genetic algorithm procedure incorporated with the optimal train control algorithm to find the optimal solution. Finally, we present a simple example and a real-world example based on the operation data from the Beijing Metro Yizhuang Line in Beijing, China. The results of the real-world example show that, during peak hours, off-peak hours and night hours, the total tractive energy consumptions can be reduced by: (1) 10.66%, 9.94% and 9.13% in comparison with the current timetable and speed profile; and (2) 3.35%, 3.12% and 3.04% in comparison with the deterministic model.

Chiral separation and determination of ketamine and norketamine in hair by capillary electrophoresis

Ketamine, traditionally available as racemic mixture, has recently become available in the form of the single S-enantiomer, due to its higher anaesthetic potency associated with faster recovery times. The different pharmaceutical forms and the different pharmacodynamics of the two enantiomers imply the need for a chiral method, since most available analytical methods for biological matrices are not enantioselective.The method herein showed consists of simple capillary zone electrophoresis (CZE) for the chiral separation of ketamine and its major metabolite, norketamine, in hair specimens. After liquid-liquid extraction, the samples were electrokinetically injected and analysed in CE (running buffer: 15mM Tris phosphate pH 2.5, containing HS-γ-CDs, 0.1%, w/v). A complete separation of both racemic ketamine and norketamine in the respective enantiomers was obtained in less than 10minutes. Limit of detection (LOD) and limit of quantification (LOQ) were 0.08ng/mg and 0.25ng/mg, respectively. Percent recovery varied from 49% to 91% for all four enantiomers. Matrix effect on spiked hair samples demonstrated values ranging from 63% to 119%. Linearity was estimated using a calibration curve consisting of five concentration levels for each enantiomer (0.5−8.0ng/mg); the regression coefficients (R2) of weighted (1/x2) linear regression were all >0.988.The method is suitable for the analysis of real-world hair samples in order to investigate ketamine chronic abuse and to discriminate between the type of abused drug, either single enantiomer or racemic drug.

Ionic liquids in enhancing the sensitivity of capillary electrophoresis: Off-line and on-line sample preconcentration techniques.

The popularity of ionic liquids (ILs) has grown during the last decade in enhancing the sensitivity of CE through different off-line or on-line sample preconcentration techniques. Water-insoluble ILs were commonly used in IL-based liquid phase microextraction, in all its variants, as off-line sample preconcentration techniques combined with CE. Water-soluble ILs were rarely used in IL-based aqueous two phase system (IL-ATPS) as an off-line sample preconcentration approach combined with CE in spite of IL-ATPS predicted features such as more compatibility with CE sample injection due to its relatively low viscosity and more compatibility with CE running buffers avoid, in some cases, anion exchange precipitation. Therefore, the attentions for the key parameters affecting the performance of IL-ATPSs were generally presented and discussed. On-line CE preconcentration techniques containing IL-based surfactants at nonmicellar or micellar concentrations have become another interesting area to improve CE sensitivity and it is likely to remain a focus of the field in the endeavor because of their numerous to create rapid, simple and sensitive systems. In this article, significant contributions of ILs in enhancing the sensitivity of CE are described, and a specific overview of the relevant examples of their applications is also given.

Restricting calcium currents is required for correct fiber type specification in skeletal muscle.

ABSTRACTSkeletal muscle excitation-contraction (EC) coupling is independent of calcium influx. In fact, alternative splicing of the voltage-gated calcium channel CaV1.1 actively suppresses calcium currents in mature muscle. Whether this is necessary for normal development and function of muscle is not known. However, splicing defects that cause aberrant expression of the calcium-conducting developmental CaV1.1e splice variant correlate with muscle weakness in myotonic dystrophy. Here, we deleted CaV1.1 (Cacna1s) exon 29 in mice. These mice displayed normal overall motor performance, although grip force and voluntary running were reduced. Continued expression of the developmental CaV1.1e splice variant in adult mice caused increased calcium influx during EC coupling, altered calcium homeostasis, and spontaneous calcium sparklets in isolated muscle fibers. Contractile force was reduced and endurance enhanced. Key regulators of fiber type specification were dysregulated and the fiber type composition was shifted toward slower fibers. However, oxidative enzyme activity and mitochondrial content declined. These findings indicate that limiting calcium influx during skeletal muscle EC coupling is important for the secondary function of the calcium signal in the activity-dependent regulation of fiber type composition and to prevent muscle disease.

Capillary electrophoresis for fast detection of heterogeneous population in colistin-resistant Gram-negative bacteria.

It has been shown that diverse strains of bacteria can be separated according to their characteristic surface properties by means of CE. We employed here this analytical technique to the study of colistin-resistance in Gram-negative bacteria, which involves the selection of mutants with modified outer membrane composition resulting in changes of surface cell properties. In the same way as with molecular entities, we performed firstly the validation of an ITP-based CE method for three common pathogenic Gram-negative bacteria namely Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae. Secondly, we compared the electrophoretic profiles of bacterial samples from a colistin-susceptible clinical isolate of K. pneumoniae and from the corresponding colistin-resistant derivative. By a simple CE run taking a few minutes, the coexistence of several bacterial subpopulations in the colistin-resistant derivative was clearly evidenced. This work encourages further research that would allow applications of CE in clinical laboratory for a daily monitoring of bacterial population in cared patients when "last-chance" colistin treatment is initiated against multidrug-resistant bacteria.

Suspending force modeling based on nonlinear acoustics and high-speed running experiments for an ultrasonic journal bearing

As a kind of promising noncontact bearings, ultrasonic bearings actuated by smart materials such as lead zirconate titanate ceramics show a good application prospect in high-speed machines and precision-measuring devices. The suspending force is one of the most important parameters that play a dominated role on the bearing’s static and dynamic performance. A suspending force model based on acoustic radiation theory for cylindrical object near sound source is built to predict the radial carrying capacity of an ultrasonic bearing actuated by three piezoelectric transducers. To validate the model, an ultrasonic bearing prototype is developed and a testing system is established. For observing the bearing’s dynamic running performance at high speeds, the bearing’s running experiment is carried out and the rotor center’s trajectory data and frequency spectrum are acquired to analyze the bearing’s dynamic characteristics at high speeds. The suspending force model and running performance experiments will contribute to the design, detection, and test of this type of bearings.

CE with chemiluminescence detection for the determination of thyroxine in human serum

A sensitive and rapid approach to perform thyroxine (T4) assay by CE with chemiluminescence (CL) detection was developed. The sensitive detection was based on the enhancement effect of T4 on the CL reaction between luminol and potassium permanganate (KMnO4 ) in alkaline solution. A laboratory-built reaction flow cell and a photon counter were deployed for the CL detection. Experimental conditions for CL detection were studied in detail to achieve maximum assay sensitivity. Optimal conditions were found to be 5.0 × 10(-4) M luminol added to the CE running buffer and 9.2 × 10(-5) M KMnO4 in 0.072 M NaOH solution introduced postcolumn. In the optimized experimental conditions, the linear range for T4 detection was 6.0 × 10(-8) -6.0 × 10(-6) M, with the detection limit of 2.0 × 10(-8) M (S/N = 3). Six human serum samples from healthy subjects, hyperthyroid patients and hypothyroid patients were analyzed by the presented method. The serum level of T4 in healthy subjects was found be 9.0 × 10(-8) M, whereas the T4 level was found to be 15.6 × 10(-8) M in hyperthyroid patients and 1.3 × 10(-8) M in hypothyroid patients. The results suggested a potential application of the proposed assay in rapid primary diagnosis of diseases such as hyperthyroid and hypothyroid.

A novel method for the analysis of 20 multi‐Indel polymorphisms and its forensic application

Insertion/deletion polymorphisms (Indels) have been considered as potential markers for forensic DNA analysis. However, the discrimination power of Indels is relatively lower due to the poor polymorphisms of diallelic markers. Here, two to three Indel loci that were very tightly linked in physical position were combined into a specific multi-Indel marker to improve the discrimination, as well as a multiplex that consisted of a set of multi-Indel markers was developed for forensic purpose. Finally, a multiplex system with 20 multi-Indel markers including 43 Indel loci from dbSNP database was constructed and DNA sample can be analyzed by this multiplex in one PCR reaction and one CE run. A total of 150 unrelated individuals from Hunan province in South-central China were genotyped by the multiplex system. The result showed that a total of 63 specific amplicons were detected, three alleles were observed in multi-Indel markers including two Indel loci and four alleles were observed in the markers including three Indel loci. The cumulative probability of exclusion and the accumulated discrimination power were 0.9989 and 0.9999999999994, respectively. Our result demonstrated that the strategy could be efficient to develop higher polymorphic multi-Indel markers, and the new multiplex could provide Supporting Information for forensic application.

Capillary zone electrophoresis and capillary electrophoresis-mass spectrometry for analyzing qualitative and quantitative variations in therapeutic albumin

The present study describes a reproducible and quantitative capillary zone electrophoresis (CZE) method, which leads to the separation of nine forms (native, oxidized and glycated) of human serum albumin (HSA). In an attempt to identify the different species separated by this CZE method, the capillary electrophoresis was coupled to mass spectrometry using a sheath liquid interface, an optimized capillary coating and a suitable CE running buffer. CE-MS analyses confirmed the heterogeneity of albumin preparation and revealed new truncated and modified forms such as Advanced Glycation End products (AGEs). Assignment of the CZE peaks was carried out using specific antibodies, carboxypeptidase A or sample reduction before or during the CE separation. Thus, five HSA forms were unambiguously identified. Using this CZE method several albumin batches produced by slightly different fractionation ways could be discriminated. Furthermore, analyses of HSA preparations marketed by five pharmaceutical industries revealed that two therapeutic albumins, including that marketed by LFB, contained the highest proportion of native form and lower levels of oxidized forms.

Amino acid analysis of dried blood spots for diagnosis of phenylketonuria using capillary electrophoresis-mass spectrometry equipped with a sheathless electrospray ionization interface

We describe a capillary electrophoresis-mass spectrometry (CE-MS) method for newborn screening of a representative amino acid metabolic disease, namely, phenylketonuria (PKU). Underivatized phenylalanine and tyrosine in a dried blood spot (DBS) were simultaneously determined by CE-MS equipped with an ionophore membrane-packed sheathless electrospray ionization interface, which was developed by our group. The method was optimized for rapid determination of the underivatized amino acids, phenylalanine and tyrosine extracted from a DBS. Under the optimized conditions, the limit of detection of phenylalanine and tyrosine (signal-to-noise ratio, 3) was 0.03 and 0.07 mg/L in DBS, respectively, with a CE run time of less than 3 min. For repeated runs of a sample, coefficients of variation (CVs) for migration time were less than 3.7%, whereas CVs for the area ratio under the curve were 2.1 and 2.9% for 20 consecutive runs of 49.5 mg/kg Phe and 36.2 mg/kg Tyr, respectively. However, the relative standard deviations of intra- and interday assays for DBS samples were <6.2 and <5.8%, respectively, which were substantially due to sample extraction from DBS. The analytical method was applied to real clinical samples of Korean neonates, and results were compared with those of conventional methods for PKU diagnosis, which required reference analytical methods such as isotope dilution CE-MS or high-performance liquid chromatography-mass spectrometry for quality assurance of the conventional kit-based assays. The distinct advantages of high sensitivity and extremely low sample volume, as well as a simple, easy, and economic sample pretreatment, were demonstrated for the proposed method.

Screen of Monoamine Oxidase Inhibitors by Protein-Liposome Conjugate Capillary Electrophoresis

Abstract Based on the interaction between enzyme and substrate, a simple and elegant method was developed to screen the inhibitors of monoamine oxidase (MAO). Four kinds of N-propargyl amine compounds with gradient concentrations were added in the CE running buffer solution containing protein-liposome conjugate to inhibit the activity of MAO to some extent. The relative migration time ratio (RMTR) of Kyn was investigated in running buffer solution containing different compounds in gradient concentrations. The results indicate that R-N-(2-heptyl)-N-methylpropargylamine (R-2-HMP) and N-propargyl-R-2-heptylamine (R-2-HPA) could inhibit the activity of MAO obviously, such that the interaction between MAO and Kyn was weakened, RMTR of Kyn increased with the increase of R-2-HMP and R-2-HPA. The inhibition activities of N-dipropargyl-R-2-hexylamine and N,N-dipropargyl-R-2-octylamine were not obvious; RMTR of Kyn changed little with the increase of the two compounds. This result was consistent with that of activity assay of compounds incubation offline. In comparison with the traditional screening method for MAO inhibitors, this method is rapid, low cost, less consumption of enzyme and independent of voltage and other interference factors.