Factors affecting DNA uptake by mammalian cells

The ability to introduce DNA and express custom DNA sequences in bacteria opened the door for improvements in a large number of fields including agriculture, pharmacology, medicine, nutrition, etc. The ability to introduce foreign DNA sequences into mammalian cells in an efficient manner would have a large impact on therapeutic applications especially gene therapy. The methods in use today suffer from low efficiencies and sometimes toxicity. In this work a number of factors were evaluated for their effect onONA uptake efficiency. The factors studied included exposure to sublethal concentration of hydrogen peroxide which have been show to lead to destabilisation ofthe lysosomes. These exposures have proven to be very toxic to cells when combined with either the calcium phosphate or the lipofectAMINE® transfection methods. Another factor evaluated was exposure to Electro-Magnetic Fields (EMF). This was fuelled by the fact that EMF have been shown to mediate a number of effects on cell structure and/or physiology. EMF exposure by itself was not sufficient to induce the cells to pick up the DNA, therefore its effect on calcium phosphate and lipofectAMINE® was tested. Although some positive results were obtained, the variability of these results exceeded by far any observed enhancements which discouraged any further work on EMF. Also tested was the possible effect the presence of the cytomegalovirus (CMV) sequence might have on DNA uptake (based on previous results in this lab). It was found that the presence ofCMV in the DNA sequence does not enhance uptake or slow down degradation of the internalised DNA. The final factor tested was the effect of basic amino acids on transfection efficiency. It was found that arginine can enhance DNA uptake by about 170% v/ith calcium phosphate and about 200% with LipofectAMINE®. A model was proposed to explain the effect of arginine as well as the lack of effect from other amino acids.

The role of daytime napping in sleepiness and cognitive function in 24-70 year olds /

Daytime napping improves well-being and performance for young adults. The benefits of napping in older adults should be investigated because they have fragmented nocturnal sleep, cognitive declines, and more opportunity to nap. In addition, experience with napping might influence the benefits of napping. Study 1 examined the role of experience with napping in young adults. Habitual (n = 23) and non-habitual nappers (n = 16) were randomly assigned to a 20-minute nap or a 20- minute reading condition. Both groups slept the same according to macro architecture. However, microarchitecture showed greater theta, alpha, and beta power during Stage 1, and greater delta, alpha, and sigma power during Stage 2 for habitual nappers, for the most part indicating better sleep. Both groups felt less sleepy after the nap. P2 latency, reflecting information processing, decreased after the nap for habitual nappers, and after the control condition for non-habitual nappers. In sum, both groups who slept felt better, but only the habitual nappers who napped gained a benefit in terms of information processing. Based on this outcome, experience with napping was investigated in Study 2. Study 2 examined the extent to which daytime napping enhanced cognition in older adults, especially frontal lobe function. Cognitive deficits in older adults may be due to sleep loss and age-related decline in brain functioning. Longer naps were expected to provide greater improvement, particularly for older adults, by reducing sleep pressure. Thirty-two adults, aged 24-70 years, participated in a repeated measures dose-response manipulation of sleep pressure. Twenty- and sixty-minute naps were compared to a no-nap condition in three age groups. Mood, subjective sleepiness, reaction time, working memory, 11 novelty detection, and waking electro physiological measures were taken before and after each condition. EEG was also recorded during each nap or rest condition. Napping reduced subjective sleepiness, improved working memory (serial addition / subtraction task), and improved attention (reduced P2 amplitude). Physiological sleepiness (i.e., waking theta power) increased following the control condition, and decreased after the longer nap. Increased beta power after the short nap, and seen with older adults overall, may have reflected increased mental effort. Older adults had longer latencies and smaller amplitudes for several event-related potential components, and higher beta and gamma power. Following the longer nap, gamma power decreased for older adults, but increased for young adults. Beta and gamma power may represent enhanced alertness or mental effort. In addition, Nl amplitude showed that benefits depend on the preceding nap length as well as age. Since the middle group had smaller Nl amplitudes following the short nap and rest condition, it is possible that they needed a longer nap to maintain alertness. Older adults did not show improvements to Nl amplitude following any condition; they may have needed a nap longer than 60 minutes to gain benefits to attention or early information processing. Sleep characteristics were not related to benefits of napping. Experience with napping was also investigated. Subjective data confirmed habitual nappers were happier to nap, while non-habitual nappers were happier to stay awake, reflecting self-identified napping habits. Non-habitual nappers were sleepier after a nap, and had faster brain activity (i.e., heightened vigilance) at sleep onset. These reasons may explain why non-habitual nappers choose not to nap.

Neuromuscular adaptations in endurance-trained boys and men

Competitive sports participation in youth is becoming increasingly more common in the Western world. It is widely accepted that sports participation, specifically endurance training, is beneficial for physical, psychomotor, and social development of children. The research on the effect of endurance training in children has focused mainly on healthrelated benefits and physiological adaptations, particularly on maximal oxygen uptake. However, corresponding research on neuromuscular adaptations to endurance training and the latter's possible effects on muscle strength in youth is lacking. In children and adults, resistance training can enhance strength and mcrease muscle activation. However, data on the effect of endurance training on strength and neuromuscular adaptations are limited. While some evidence exists demonstrating increased muscle activation and possibly increased strength in endurance athletes compared with untrained adults, the neuromuscular adaptations to endurance training in children have not been examined. Thus, the purpose of this study was to examine maximal isometric torque and rate of torque development (RID), along with the pattern of muscle activation during elbow and knee flexion and extension in muscle-endurancetrained and untrained men and boys. Subjects included 65 males: untrained boys (n=18), endurance-trained boys (n=12), untrained men (n=20) and endurance-trained men (n=15). Maximal isometric torque and rate of torque development were measured using an isokinetic dynamometer (Biodex III), and neuromuscular activation was assessed using surface electromyography (SEMG). Muscle strength and activation were assessed in the dominant arm and leg, in a cross-balanced fashion during elbow and knee flexion and extension. The main variables included peak torque (T), RTD, rate of muscle activation (Q30), Electro-mechanical delay (EMD), time to peak RTD and co-activation index. Age differences in T, RTD, electro-mechanical delay (EMD) and rate of muscle activation (Q30) were consistently observed in the four contractions tested. Additionally, Q30, nonnalized for peak EMG amplitude, was consistently higher in the endurancetrained men compared with untrained men. Co-activation index was generally low in all contractions. For example, during maximal voluntary isometric knee extension, men were stronger, had higher RTD and Q30, whether absolute or nonnalized values were used. Moreover, boys exhibited longer EMD (64.8 ± 18.5 ms vs. 56.6 ± 15.3 ms, for boys and men respectively) and time to peak RTD (112.4 ± 33.4 ms vs. 100.8 ± 39.1 ms for boys and men, respectively). In addition, endurance-trained men had lower T compared with untrained men, yet they also exhibited significantly higher nonnalized Q30 (1.9 ± 1.2 vs. 1.1 ± 0.7 for endurance-trained men and untrained men, respectively). No training effect was apparent in the boys. In conclusion, the findings demonstrate muscle strength and activation to be lower in children compared with adults, regardless of training status. The higher Q30 of the endurance-trained men suggests neural adaptations, similar to those expected in response to resistance training. The lower peak torque may su9gest a higher relative involvement oftype I muscle fibres in the endurance-trained athletes. Future research is required to better understand the effect of growth and development on muscle strength and activation patterns during dynamic and sub-maximal isometric contractions. Furthennore, training intervention studies could reveal the effects of endurance training during different developmental stages, as well as in different muscle groups.