Three methods for examining the de novo proteome of microglia using BONCAT bioorthogonal labeling and FUNCAT click chemistry.

Bioorthogonal labeling and click chemistry techniques allow the detailed examination of cellular physiology through tagging and visualizing newly synthesized proteins. Here, we describe three methods applying bioorthogonal non-canonical amino acid tagging and fluorescent non-canonical amino acid tagging to quantify protein synthesis in microglia. We describe steps for cell seeding and labeling. We then detail microscopy, flow cytometry, and Western blotting techniques. These methods can be easily adapted for other cell types to explore cellular physiology in health and disease. For complete details on the use and execution of this protocol, please refer to Evans et al. (2021).1.

Neurogenic-dependent changes in hippocampal circuitry underlie the procognitive effect of exercise in aging mice.

We have shown that the improvement in hippocampal-based learning in aged mice following physical exercise observed is dependent on neurogenesis in the dentate gyrus (DG) and is regulated by changes in growth hormone levels. The changes in neurocircuitry, however, which may underlie this improvement, remain unclear. Using in vivo multimodal magnetic resonance imaging to track changes in aged mice exposed to exercise, we show the improved spatial learning is due to enhanced DG connectivity, particularly the strengthening of the DG-Cornu Ammonis 3 and the DG-medial entorhinal cortex connections in the dorsal hippocampus. Moreover, we provide evidence that these changes in circuitry are dependent on neurogenesis since they were abrogated by ablation of newborn neurons following exercise. These findings identify the specific changes in hippocampal circuitry that underlie the cognitive improvements resulting from physical activity and show that they are dependent on the activation of neurogenesis in aged animals.

An exercise "sweet spot" reverses cognitive deficits of aging by growth-hormone-induced neurogenesis.

Hippocampal function is critical for spatial and contextual learning, and its decline with age contributes to cognitive impairment. Exercise can improve hippocampal function, however, the amount of exercise and mechanisms mediating improvement remain largely unknown. Here, we show exercise reverses learning deficits in aged (24 months) female mice but only when it occurs for a specific duration, with longer or shorter periods proving ineffective. A spike in the levels of growth hormone (GH) and a corresponding increase in neurogenesis during this sweet spot mediate this effect because blocking GH receptor with a competitive antagonist or depleting newborn neurons abrogates the exercise-induced cognitive improvement. Moreover, raising GH levels with GH-releasing hormone agonist improved cognition in nonrunners. We show that GH stimulates neural precursors directly, indicating the link between raised GH and neurogenesis is the basis for the substantially improved learning in aged animals.

The effect of alterations in foot centre of pressure on lower body kinematics during the five-iron golf swing.

The research aimed to evaluate the effects of an intervention aimed at altering pressure towards the medial aspect of the foot relating to stability mechanisms associated with the golf swing. We hypothesised that by altering the position of the foot pressure, the lower body stabilisation would improve which in turn would enhance weight distribution and underpinning lower body joint kinematics. Eight professional golf association (PGA) golf coaches performed five golf swings, recorded using a nine-camera motion analysis system synchronised with two force platforms. Following verbal intervention, they performed further five swings. One participant returned following a one-year intervention programme and performed five additional golf swings to provide a longitudinal case study analysis. Golf performance was unchanged evidenced by the velocity and angle of the club at ball impact (BI), although the one-year intervention significantly changed the percentage of weight experienced at each foot in the final 9% of downswing, which provided an even weight distribution at BI. This is a highly relevant finding as it indicates that the foot centre of pressure was central to the base of support and in-line with the centre of mass (CoM), indicating significantly increased stability when the CoM is near maximal acceleration.

Testing hypotheses of developmental constraints on mammalian brain partition evolution, using marsupials.

There is considerable debate about whether the partition volumes of the mammalian brain (e.g. cerebrum, cerebellum) evolve according to functional selection, or whether developmental constraints of conserved neurogenetic scheduling cause predictable partition scaling with brain size. Here we provide the first investigation of developmental constraints on partition volume growth, derived from contrast-enhanced micro-computed tomography of hydrogel-stabilized brains from three marsupial species. ANCOVAs of partition vs. brain volume scaling, as well as growth curve comparisons, do not support several hypotheses consistent with developmental constraints: brain partition growth significantly differs between species, or between developing vs. adult marsupials. Partition growth appears independent of adult brain volume, with no discernable growth spurts/lags relatable to internal structural change. Rather, adult proportion differences appear to arise through growth rate/duration heterochrony. Substantial phylogenetic signal in adult brain partitions scaling with brain volume also counters expectations of development-mediated partition scaling conservatism. However, the scaling of olfactory bulb growth is markedly irregular, consistent with suggestions that it is less constrained. The very regular partition growth curves suggest intraspecific developmental rigidity. We speculate that a rigid, possibly neuromer-model-like early molecular program might be responsible both for regular growth curves within species and impressions of a link between neurogenesis and partition evolution.

A High Omega-3 Fatty Acid Multinutrient Supplement Benefits Cognition and Mobility in Older Women: A Randomized, Double-blind, Placebo-controlled Pilot Study.

BACKGROUND: Mobility is a key determinant of frailty in older persons, and a variety of dietary factors, such as the omega-3 fatty acid docosahexaenoic acid (DHA), are positively associated with decreased frailty and improved mobility and cognition in older persons. METHODS: The effects of a multinutrient supplement on mobility and cognition were assessed in postmenopausal women (60-84 years). Participants received either Efalex Active 50+ (1g DHA, 160 mg eicosapentaenoic acid, 240 mg Ginkgo biloba, 60 mg phosphatidylserine, 20mg d-α tocopherol, 1mg folic acid, and 20 µg vitamin B12 per day; N = 15) or placebo (N = 12) for 6 months. Mobility was assessed by VICON 9 motion capture camera system synchronized with Kistler force plates, cognitive performance by computerized cognitive function tests, and blood fatty acid levels by pin-prick analysis. RESULTS: Significant effects of treatment were seen in two of the four cognitive tests, with shorter mean latencies in a motor screening task (p < .05) and more words remembered (p < .03), and one of the three primary mobility measures with improved habitual walking speed (p < .05). Compared with the placebo group, supplementation also resulted in significantly higher blood DHA levels (p < .02). CONCLUSIONS: In this pilot study, multinutrient supplementation improved cognition and mobility in able older females at clinically relevant levels, suggesting a potential role in reducing the decline to frailty.

Detection of falls using accelerometers and mobile phone technology.

OBJECTIVES: to study the sensitivity and specificity of fall detection using mobile phone technology. DESIGN: an experimental investigation using motion signals detected by the mobile phone. SETTING AND PARTICIPANTS: the research was conducted in a laboratory setting, and 18 healthy adults (12 males and 6 females; age = 29 ± 8.7 years) were recruited. MEASUREMENT: each participant was requested to perform three trials of four different types of simulated falls (forwards, backwards, lateral left and lateral right) and eight other everyday activities (sit-to-stand, stand-to-sit, level walking, walking up- and downstairs, answering the phone, picking up an object and getting up from supine). Acceleration was measured using two devices, a mobile phone and an independent accelerometer attached to the waist of the participants. RESULTS: Bland-Altman analysis shows a higher degree of agreement between the data recorded by the two devices. Using individual upper and lower detection thresholds, the specificity and sensitivity for mobile phone were 0.81 and 0.77, respectively, and for external accelerometer they were 0.82 and 0.96, respectively. CONCLUSION: fall detection using a mobile phone is a feasible and highly attractive technology for older adults, especially those living alone. It may be best achieved with an accelerometer attached to the waist, which transmits signals wirelessly to a phone.