Dielectrophoretic differentiation of mouse ovarian surface epithelial cells, macrophages, and fibroblasts using contactless dielectrophoresis

Ovarian cancer is the leading cause of death from gynecological malignancies in women. The primary challenge is the detection of the cancer at an early stage, since this drastically increases the survival rate. In this study we investigated the dielectrophoretic responses of progressive stages of mouse ovarian surface epithelial (MOSE) cells, as well as mouse fibroblast and macrophage cell lines, utilizing contactless dielectrophoresis (cDEP). cDEP is a relatively new cell manipulation technique that has addressed some of the challenges of conventional dielectrophoretic methods. To evaluate our microfluidic device performance, we computationally studied the effects of altering various geometrical parameters, such as the size and arrangement of insulating structures, on dielectrophoretic and drag forces. We found that the trapping voltage of MOSE cells increases as the cells progress from a non-tumorigenic, benign cell to a tumorigenic, malignant phenotype. Additionally, all MOSE cells display unique behavior compared to fibroblasts and macrophages, representing normal and inflammatory cells found in the peritoneal fluid. Based on these findings, we predict that cDEP can be utilized for isolation of ovarian cancer cells from peritoneal fluid as an early cancer detection tool.     

Mapping of hyperthermic tumor cell death in a microchannel under unidirectional heating

Hyperthermia can be used as an adjunctive method of chemotherapy, radiotherapy, and gene therapy to improve cancer treatment. In this study, we investigate the hyperthermic cell death of cervix cancer CaSki cells in a microchannel integrated with a directional heating scheme. Heat was applied from the inner end to the outer end of the channel and a temperature distribution from 60 °C to 30 °C was established. A three dimensional (3D) numerical model was conducted for the heat transfer simulation, based on which a simple fitting method was proposed to easily estimate the temperature distribution along the channel. Cell death along the channel was mapped 22 h after the heating treatment by dual fluorescent labeling and phase-contrast microscopy imaging. Upstream, where the temperature is higher than 42 °C, we observe necrotic death, late-stage and early stage apoptotic death in sequence along the channel. Downstream and in the middle of the channel, where the temperature is lower than 42 °C, significant cell detachment was noted. Vigorous detachment was observed even in the non-hyperthermic zone (temperature lower than 37 °C), which we believe is due to the direct effect of the hyperthermic zones (higher than 37 °C). The present work not only gives a vivid map of cell responses under a temperature gradient, but also reveals the potential interactions of the heated tumor cells and non-heated tumor cells, which are seldom investigated in conventional petri-dish experiments.     

Formation of embryoid bodies using dielectrophoresis

Embryoid body (EB) formation forms an important step in embryonic stem cell differentiation invivo. In murine embryonic stem cell (mESC) cultures EB formation is inhibited by the inclusion of leukaemic inhibitory factor (LIF) in the medium. Assembly of mESCs into aggregates by positive dielectrophoresis (DEP) in high field regions between interdigitated oppositely castellated electrodes was found to initiate EB formation. Embryoid body formation in aggregates formed with DEP occurred at a more rapid rate-in fact faster compared to conventional methods-in medium without LIF. However, EB formation also occurred in medium in which LIF was present when the cells were aggregated with DEP. The optimum characteristic size for the electrodes for EB formation with DEP was found to be 75-100 microns; aggregates smaller than this tended to merge, whilst aggregates larger than this tended to split to form multiple EBs. Experiments with ESCs in which green fluorescent protein (GFP) production was targeted to the mesodermal gene brachyury indicated that differentiation within embryoid bodies of this size may preferentially occur along the mesoderm lineage. As hematopoietic lineages during normal development derive from mesoderm, the finding points to a possible application of DEP formed EBs in the production of blood-based products from ESCs.     

Controlled electroporation of the plasma membrane in microfluidic devices for single cell analysis

Chemical cytometry on a single cell level is of interest to various biological fields ranging from cancer to stem cell research. The impact chemical cytometry can exert in these fields depends on the dimensionality of the retrievable analytes content. To this point, the number of different analytes identifiable and additionally their subcellular localization is of interest. To address this, we present an electroporation based approach for selective lysis of only the plasma membrane, which permits analysis of the dissolved cytoplasm, while reducing contributions from the nucleus and membrane bound fractions of the cell analytes. The use of 100 μs long pulse and a well defined DC electric field gradient of ∼4.5 kV·cm⁻¹ generated by 3D electrodes initiates release of a cytoplasm marker in ≪1 s, while retaining nuclear fluorescence markers.     

Intra-task variability of trunk coordination during a rate-controlled bipedal dance jump

In this study, we investigated trunk coordination during rate-controlled bipedal vertical dance jumps. The aims of the study were to investigate the pattern of coordination and the magnitude of coordination variability within jump phases and relative to phase-defining events during the jump. Lumbar and thoracic kinematics were collected from seven dancers during a series of jumps at 95 beats per minute. The vector coding technique was used to quantify the pattern and variability of trunk coordination. Coordination was predominantly anti-phase during propulsion and landing. Mean coordination variability peaked just before the landing phase and at the transition from landing to propulsion phases, and was lowest during the propulsion phase just before toe-off. The results indicate that peaks in variability could be explained by task and phase-specific biomechanical demands.     

Repeated exercise stress impairs volitional but not magnetically evoked electromechanical delay of the knee flexors

The effects of serial episodes of fatigue and recovery on volitional and magnetically evoked neuromuscular performance of the knee flexors were assessed in 20 female soccer players during: (i) an intervention comprising 4 × 35 s maximal static exercise, and (ii) a control condition. Volitional peak force was impaired progressively (-16% vs. baseline: 235.3 ± 54.7 to 198.1 ± 38.5 N) by the fatiguing exercise and recovered to within -97% of baseline values following 6 min of rest. Evoked peak twitch force was diminished subsequent to the fourth episode of exercise (23.3%: 21.4 ± 13.8 vs. 16.4 ± 14.6 N) and remained impaired at this level throughout the recovery. Impairment of volitional electromechanical delay performance following the first episode of exercise (25.5%: 55.3 ± 11.9 vs. 69.5 ± 24.5 ms) contrasted with concurrent improvement (10.0%: 24.5 ± 4.7 vs. 22.1 ± 5.0 ms) in evoked electromechanical delay (P < 0.05), and this increased disparity between evoked and volitional electromechanical delay remained during subsequent periods of intervention and recovery. The fatiguing exercise provoked substantial impairments to volitional strength and volitional electromechanical delay that showed differential patterns of recovery. However, improved evoked electromechanical delay performance might identify a dormant capability for optimal muscle responses during acute stressful exercise and an improved capacity to maintain dynamic joint stability during critical episodes of loading.     

Influence of a conservative sleep management strategy during a solo Pacific Ocean crossing on anxiety and perceived fatigue: a case study

The aim of this case study was to determine whether a sailor's deliberate choice of a conservative strategy to manage sleep deprivation would allow him to cross the Pacific Ocean and to minimize his state of anxiety and perceived fatigue. The participant, who had more than 10 years' sailing experience in severe conditions, was tested on a small catamaran without any living quarters during a solo Pacific Ocean crossing. Estimations of sleep hours, state anxiety, and perceived fatigue were self-reported by the sailor on a daily basis using a specific questionnaire. The most important finding is that the sailor's deliberate sleep strategy, 5.4 h sleep per day (24% less than on-shore), was enough to keep his anxiety and perceived fatigue within acceptable limits and enabled him to achieve his goal, which was the first crossing of the Pacific Ocean on a catamaran of less than 6 m. In conclusion, our results suggest that the sailor observed in the present case study was able to minimize anxiety and perceived fatigue with adequate sleep to optimize his performance, security, and to achieve his goal.     

Mixed-method pre-cooling reduces physiological demand without improving performance of medium-fast bowling in the heat

This study examined physiological and performance effects of pre-cooling on medium-fast bowling in the heat. Ten, medium-fast bowlers completed two randomised trials involving either cooling (mixed-methods) or control (no cooling) interventions before a 6-over bowling spell in 31.9±2.1°C and 63.5±9.3% relative humidity. Measures included bowling performance (ball speed, accuracy and run-up speeds), physical characteristics (global positioning system monitoring and counter-movement jump height), physiological (heart rate, core temperature, skin temperature and sweat loss), biochemical (serum concentrations of damage, stress and inflammation) and perceptual variables (perceived exertion and thermal sensation). Mean ball speed (114.5±7.1 vs. 114.1±7.2 km · h(-1); P = 0.63; d = 0.09), accuracy (43.1±10.6 vs. 44.2±12.5 AU; P = 0.76; d = 0.14) and total run-up speed (19.1±4.1 vs. 19.3±3.8 km · h(-1); P = 0.66; d = 0.06) did not differ between pre-cooling and control respectively; however 20-m sprint speed between overs was 5.9±7.3% greater at Over 4 after pre-cooling (P = 0.03; d = 0.75). Pre-cooling reduced skin temperature after the intervention period (P = 0.006; d = 2.28), core temperature and pre-over heart rates throughout (P = 0.01-0.04; d = 0.96-1.74) and sweat loss by 0.4±0.3 kg (P = 0.01; d = 0.34). Mean rating of perceived exertion and thermal sensation were lower during pre-cooling trials (P = 0.004-0.03; d = 0.77-3.13). Despite no observed improvement in bowling performance, pre-cooling maintained between-over sprint speeds and blunted physiological and perceptual demands to ease the thermoregulatory demands of medium-fast bowling in hot conditions.     

The effects of Ramadan intermittent fasting on athletic performance: recommendations for the maintenance of physical fitness

The behavioural modifications that accompany Ramadan intermittent fasting (RIF) are usually associated with some alterations in the metabolic, physiological, and psychological responses of athletes that may affect sport performance. Muslim athletes who are required to train and/or compete during the month-long, diurnal fast must adopt coping strategies that allow them to maintain physical fitness and motivation if they are to perform at the highest level. This updated review aims to present the current state of knowledge of the effects of RIF on training and performance, focusing on key-factors that contribute to the effects of Ramadan on exercise performance: energy restriction, sleep deprivation, circadian rhythm perturbation, dehydration, and alterations in the training load. The available literature contain few studies that have examined the effects of RIF on physical performance in athletes and, to date, the results are inconclusive, so the effects of RIF on competition outcomes are not at present wholly understood. The diverse findings probably indicate individual differences in the adaptability and self-generated coping strategies of athletes during fasting and training. However, the results of the small number of well-controlled studies that have examined the effects of Ramadan on athletic performance suggest that few aspects of physical fitness are negatively affected, and where decrements are observed these are usually modest. Subjective feelings of fatigue and other mood indicators are often cited as implying additional stress on the athlete throughout Ramadan, but most studies show that these factors may not result in decreases in performance and that perceived exercise intensity is unlikely to increase to any significant degree. Current evidence from good, well-controlled research supports the conclusion that athletes who maintain their total energy and macronutrient intake, training load, body composition, and sleep length and quality are unlikely to suffer any substantial decrements in performance during Ramadan. Further research is required to determine the effect of RIF on the most challenging events or exercise protocols and on elite athletes competing in extreme environments.     

Comparison of power and EMG during 6-s all-out cycling between young and older women

Abstract To investigate the effects of ageing on the neural control strategies governing sprint cycling on a friction-loaded cycle-ergometer, 10 older (aged 70-83yr) and 8 young (aged 19-35yr) healthy women completed seven 6-s all-out cycling trials against varying loads. Root mean square (RMS), median frequency and muscle fibre conduction velocity were determined from the vastus lateralis of the dominant limb during each pedal stroke. Peak power was 43% lower in the older group compared to the younger (p??0.05). ΔRMS from the first to the sixth second during each trial was found to increase significantly with the development of power output in both groups (p?相似文献