Effects of amyloid precursor protein peptide APP96-110, alone or with human mesenchymal stromal cells, on recovery after spinal cord injury.

Delivery of a peptide (APP96-110), derived from amyloid precursor protein (APP), has been shown to elicit neuroprotective effects following cerebral stroke and traumatic brain injury. In this study, the effect of APP96-110 or a mutant version of this peptide (mAPP96-110) was assessed following moderate (200 kdyn, (2 N)) thoracic contusive spinal cord injury (SCI) in adult Nude rats. Animals received a single tail vein injection of APP96-110 or mAPP96-110 at 30 minutes post-SCI and were then assessed for functional improvements over the next 8 weeks. A cohort of animals also received transplants of either viable or non-viable human mesenchymal stromal cells (hMSCs) into the SC lesion site at one week post-injury to assess the effect of combining intravenous APP96-110 delivery with hMSC treatment. Rats were perfused 8 weeks post-SCI and longitudinal sections of spinal cord analyzed for a number of factors including hMSC viability, cyst size, axonal regrowth, glial reactivity and macrophage activation. Analysis of sensorimotor function revealed occasional significant differences between groups using Ladderwalk or Ratwalk tests, however there were no consistent improvements in functional outcome after any of the treatments. mAPP96-110 alone, and APP96-110 in combination with both viable and non-viable hMSCs significantly reduced cyst size compared to SCI alone. Combined treatments with donor hMSCs also significantly increased ßIII tubulin+, glial fibrillary acidic protein (GFAP+) and laminin+ expression, and decreased ED1+ expression in tissues. This preliminary study demonstrates that intravenous delivery of APP96-110 peptide has selective, modest neuroprotective effects following SCI, which may be enhanced when combined with hMSC transplantation. However, the effects are less pronounced and less consistent compared to the protective morphological and cognitive impact that this same peptide has on neuronal survival and behaviour after stroke and traumatic brain injury. Thus while the efficacy of a particular therapeutic approach in one CNS injury model may provide justification for its use in other neurotrauma models, similar outcomes may not necessarily occur and more targeted approaches suited to location and severity are required. All animal experiments were approved by The University of Western Australia Animal Ethics Committee (RA3/100/1460) on April 12, 2016.

Patients with type 1 diabetes mellitus have impaired IL-1ß production in response to Mycobacterium tuberculosis.

Patients with diabetes mellitus have an increased risk of developing tuberculosis. Although the underlying mechanism is unclear, evidence suggests a role for chronic hyperglycaemia. We examined the influence of hyperglycaemia on Mycobacterium tuberculosis-induced cytokine responses in patients with type 1 diabetes mellitus (T1D). Peripheral blood mononuclear cells (PBMCs) from 24 male T1D patients with sub-optimal glucose control [HbA1c > 7.0% (53 mmol/L)] and from 24 age-matched male healthy controls were stimulated with M. tuberculosis lysate. Cytokine analysis, assessment of aerobic glycolysis, receptor recognition and serum cross-over experiments were performed to explore the mechanistic differences. PBMCs from T1D patients produced less bioactive interleukin (IL)-1ß in response to M. tuberculosis. IL-6 and interferon (IFN)-γ production trended towards a decrease, whilst other cytokines such as tumour necrosis factor (TNF)-α, IL-17 and IL-1Ra were normal. The decrease in cytokine production was not correlated to HbA1c or plasma glucose levels. Cross-over serum experiments did not alter the cytokine profile of T1D or control patients, arguing for an intrinsic cellular defect. Cellular metabolism and the expression of M. tuberculosis-related pattern recognition receptors (PRRs) such as TLR2, TLR4 and NOD2 did not differ between T1D patients and healthy controls. Compared to matched controls, T1D patients have a reduced capacity to produce pro-inflammatory cytokines in response to M. tuberculosis. The impaired IL-1ß production in T1D patients may contribute to the increased susceptibility to tuberculosis. This effect appears not to be related to prevailing glucose levels but to an intrinsic cellular deficit.

Characterization of an 'Amyloid Only' Transgenic (B6C3-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax) Mouse Model of Alzheimer's Disease.

The spatiotemporal pattern of cerebral amyloid deposition, detectable as light microscopically recognizable aggregates in an 'amyloid only' transgenic mouse model of Alzheimer's disease, B6C3-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax, is reported for the first time in this strain. Monoclonal and polyclonal antibodies were used to detect amyloid deposition immunohistochemically in brains collected from these mice at 3-12 months of age. Amyloid aggregates (20-200 µm) were first found in serial, whole coronal sections of brain at 4 months of age and these increased progressively, plateauing at 11-12 months. They were most abundant in the cerebral cortices, hippocampus, olfactory bulbs, some white matter tracts and the cerebellar molecular layer; no amyloid aggregates were found in the midbrain, brainstem or spinal cord, or in an equivalent number of brains from wild-type mice. Since the parahippocampal gyrus is severely damaged early in the clinical course of human Alzheimer's disease, amyloid aggregates were also assessed in this brain region and a similar temporal course of amyloid deposition was observed. Moreover, in this gyrus, the amount of aggregated amyloid showed no significant difference between left- and right-sided gyri. However, the polyclonal antibody detected a significantly greater amyloid burden than the monoclonal antibody at 3-10 months of age and the reverse was seen at 11-12 months of age. The pattern of amyloid deposition in the parahippocampal gyrus also resembled that found in the entire brain over time, when the latter was quantified by the colour deconvolution method, suggesting that this gyrus is a good marker for more widely distributed cerebral amyloid deposition. This neuropathological characterization will permit better use of the B6C3-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax transgenic mouse strain in future studies of Alzheimer's disease pathogenesis, prevention and treatment.

Psychometric properties of an omnibus sleep problems questionnaire for school-aged children.

OBJECTIVE: There is a paucity of sleep questionnaires that have been psychometrically validated for use in school-aged children. Due to the limitation regarding the psychometric properties and the great variety in question design, there remains a need for a robust omnibus questionnaire that assesses sleep problems in community populations. This study aimed to develop such a questionnaire for school-aged children by assessing the construct validity and reliability of a questionnaire based on a combination of children's sleep domains from two frequently used and validated questionnaires (Habits Questionnaire and Sleep Disorders Scale for Children) and author devised questions. PATIENTS/METHODS: Parents of 1904 children aged 5-10 years (mean 7.7 ± 1.7 years) from 32 elementary schools in Adelaide, South Australia, completed the questionnaire. RESULTS: Principal axis factoring revealed six unique sub-scales--Sleep Routine, Bedtime Anxiety, Morning Tiredness, Night Arousals, Sleep Disordered Breathing, and Restless Sleep--containing a total of 26 items. Internal consistency for sub-scales were moderate to strong (range α = 0.6-0.8) and test-retest reliability was adequate (>0.4). T-score cut-offs were devised for age and sex. CONCLUSION: The new questionnaire provides a robust set of sleep problem sub-scales which can be used for assessment of sleep concerns in a community sample as well as provide for optimal analysis of associations with other measures of childhood daytime functioning such as neurocognition and behaviour.

Prevalence of snoring and associated factors in infancy.

INTRODUCTION: In children aged 3-12 years snoring is associated with significant neurocognitive and behavioural deficits; however, there are few studies that have considered both the prevalence of snoring in infants and associated factors that may influence the development of snoring. The goal of the present study was to examine sleep, snoring and associated factors in a community sample of 0-3 month olds. METHODS: Previously validated infant sleep and parent sleep questionnaires were completed by parents of 457 term infants aged 1-13.9 weeks old (mean age=4.6 weeks; SD=2.7; 45% males) during a home-based nurse visit. RESULTS: Approximately 9% of infants were reported to snore habitually (snoring ≥ 3 nights/week). Habitual snoring was significantly associated with exclusive formula feeding (OR: 28.87; p<.01), maternal concern about child's breathing during sleep (OR: 3.91; p=.01) and restless sleep ≥ 3 nights/week (OR: 17.76; p<.001). CONCLUSION: These results show that snoring is as common in infants as it is in older children. Given the known relationships between Sleep Disordered Breathing (SDB) and neurocognitive development, the effect of SDB developing early in childhood may have important consequences on future developmental outcomes.

Chasing the silver bullet: measuring driver fatigue using simple and complex tasks.

Driver fatigue remains a significant cause of motor-vehicle accidents worldwide. New technologies are increasingly utilised to improve road safety, but there are no effective on-road measures for fatigue. While simulated driving tasks are sensitive, and simple performance tasks have been used in industrial fatigue management systems (FMS) to quantify risk, little is known about the relationship between such measures. Establishing a simple, on-road measure of fatigue, as a fitness-to-drive tool, is an important issue for road safety and accident prevention, particularly as many fatigue related accidents are preventable. This study aimed to measure fatigue-related performance decrements using a simple task (reaction time - RT) and a complex task (driving simulation), and to determine the potential for a link between such measures, thus improving FMS success. Fifteen volunteer participants (7 m, 8 f) aged 22-56 years (mean 33.6 years), underwent 26 h of supervised wakefulness before an 8h recovery sleep opportunity. Participants were tested using a 30-min interactive driving simulation test, bracketed by a 10-min psychomotor vigilance task (PVT) at 4, 8, 18 and 24h of wakefulness, and following recovery sleep. Extended wakefulness caused significant decrements in PVT and driving performance. Although these measures are clearly linked, our analyses suggest that driving simulation cannot be replaced by a simple PVT. Further research is needed to closely examine links between performance measures, and to facilitate accurate management of fitness to drive, which requires more complex assessments of performance than RT alone.

Effects of moderate sleep deprivation and low-dose alcohol on driving simulator performance and perception in young men.

STUDY OBJECTIVE: To determine the combined effects of sleep restriction and low-dose alcohol on driving simulator performance, EEG, and subjective levels of sleepiness and performance in the mid-afternoon. DESIGN: Repeated measures with 4 experimental conditions. Normal sleep without alcohol, sleep restriction alone (4 hours) and sleep restriction in combination with 2 different low blood alcohol concentrations (0.025 g/dL and 0.035 g/dL). SETTING: Sleep Laboratory, Adelaide Institute for Sleep Health. PARTICIPANTS: Twenty-one healthy young men, aged 18-30 years, mean (+/-SD) = 22.5(+/-3.7) years, BMI = 25(+/-6.7) kg/m2; all had normal sleep patterns and were free of sleep disorders. MEASUREMENTS: Participants completed a 70-minute simulated driving session, commencing at 14:00. Driving parameters included steering deviation, braking reaction time, and number of collisions. Alpha and theta EEG activity and subjective driving performance and sleepiness were also measured throughout the driving task. RESULTS: All measures were significantly affected by time. Steering deviation increased significantly when sleep restriction was combined with the higher dose alcohol. This combination also resulted in a significant increase in alpha/theta EEG activity throughout the drive, as well as greater subjective sleepiness and negative driving performance ratings compared to control or sleep restriction alone. DISCUSSION: These data indicate that combining low-dose alcohol with moderate sleep restriction results in significant decrements to subjective alertness and performance as well as to some driving performance and EEG parameters. This highlights the potential risks of driving after consumption of low and legal doses of alcohol when also sleep restricted.

Intentional and unintentional contributions to nonspecific preparation during reaction time foreperiods.

The nonspecific preparation that follows a warning stimulus (WS) to speed responding to an impending imperative stimulus (IS) is generally viewed as a strategic, intentional process. An alternative view holds that WS acts as a conditioned stimulus that unintentionally elicits a tendency to respond at the moment of IS presentation as a result of a process of trace conditioning. These views were contrasted as explanatory frameworks for classical effects on reaction time of the duration and intertrial variability of the foreperiod, the interval between WS and IS. It is shown that the conditioning view accounts for the available data at least as well as the strategic view. In addition, the results of 3 experiments provide support for the conditioning view by showing that unintentional contributions to nonspecific preparation can be dissociated from intentional contributions.

Effect of impact on different regions of the head of lambs.

The heads of anaesthetized lambs aged 4-5 weeks were subjected to impact (temporal, frontal or occipital) of constant strength with a humane stunner. Two hours later, the brains were perfusion-fixed with 4% paraformaldehyde and serial whole coronal slices processed by routine methods. Sections were stained with haematoxylin and eosin or labelled with a monoclonal antibody to amyloid precursor protein, a sensitive marker of axonal injury and neuronal reaction. Microscopical evaluation of axonal, neuronal and vascular damage was performed with a quantitative grid system. Frontal impact produced the greatest damage, followed by occipital then temporal impact. An unusual lesion found in the majority of lambs subjected to impact was multifocal necrosis of the cerebellar granular layer. The findings should assist clinicians in evaluating the probable outcome of traumatic head injury in domestic animals.

Upregulation of neuronal amyloid precursor protein (APP) and APP mRNA following magnesium sulphate (MgSO4) therapy in traumatic brain injury.

The aim of this study was to assess and quantitate topographically the effects of posttraumatic intravenous magnesium sulphate (MgSO4) on neuronal perikaryal APP antigen and messenger RNA (mRNA) expression in sheep brains 2 h after a controlled focal head impact. The percentage brain area with APP immunoreactive neuronal perikarya was 71, 56, 27.5 and 5.5%, respectively, in MgSO4-treated head-injured animals, head-injured animals without any treatment, MgSO4 treated nonimpacted animals, and nontreated nonimpacted control sheep. Although there was no statistically significant difference in APP immunoreactive neuronal perikarya in the MgSO4-treated HI group (mean 71%) compared to the HI group without any treatment (mean 56%), northern analysis showed that there was a 2.3-+/-0.2-fold increase in APP mRNA in the thalamus of treated impacted animals compared to untreated impacted animals (p < 0.005). However, MgSO4 treated nonimpacted control animals also showed a 1.6-+/-0.1-fold increase in APP mRNA compared to untreated nonimpacted controls (p < 0.005). MgSO4 therapy results in upregulation of neuronal APP mRNA and APP expression that is quantitatively greater following a focal head impact.

Urinary 6-sulfatoxymelatonin excretion and aging: new results and a critical review of the literature.

The apparent age-related decline in melatonin production has been thought to continue in a secular manner across the lifespan. While it is clear that melatonin levels in children and adolescents are elevated compared to older individuals, the question of whether there is a sudden or gradual change has not been adequately addressed. In this study, we report the excretion of the melatonin metabolite, 6-sulfatoxymelatonin in 253 subjects aged between 21 and 82 yr. The correlation with age was significant (r = -0.24; P < 0.05). When the data was analysed by ANOVA using 5-yr age spans, there was a significant effect of age, but post hoc analysis indicated that after 25 yr of age there was no significant decline in excretion of the metabolite. Thus, although the oldest subjects excreted 36% less melatonin metabolite than the youngest, the decrease occurred at a very early age. In the second part of the study, we re-evaluated the data from seven previous studies that measured plasma melatonin levels or metabolite excretion across a wide range of ages and 11 studies comparing young versus older subjects. Statistical analysis by ANOVA again suggested that the changes in melatonin occurring with age were essentially complete before 30 yr of age. The youngest subjects produced at the most twice the amount of melatonin as the oldest subjects. Finally, we evaluated the mean plasma melatonin levels in 144 groups of normal subjects reported in 137 separate publications with respect to age. Again, whereas there was a significant correlation with age, ANOVA showed that there was no difference between groups after 35 yr of age, and the oldest groups had levels that were only 43% of the youngest groups. We conclude that melatonin production is lower in older people, but that the change occurs very early in life, around 20-30 yr of age.

Upregulation of amyloid precursor protein messenger RNA in response to traumatic brain injury: an ovine head impact model.

There is evidence that the amyloid precursor protein (APP) plays an important role in neuronal growth and synaptic plasticity and that its increased expression following traumatic brain injury represents an acute phase response to trauma. We hypothesized that the previously described increased APP expression in response to injury (Van den Heuvel et al., Acta Neurochir. Suppl. 71, 209-211) is due to increased mRNA expression and addressed this by examining the expression of APP mRNA and APP within neuronal cell bodies over time in an ovine head impact model. Twenty-five anesthetized and ventilated 2-year-old Merino ewes sustained a left temporal head impact using a humane stunner and 9 normal sheep were used as nonimpact controls. Following postimpact survival periods of 15, 30, 45, 60, and 120 min, brains were perfusion fixed in 4% paraformaldehyde and examined according to standard neuropathological protocol. APP mRNA and antigen expression were examined in 5-microm sections by nonisotopic in situ hybridization and APP immunocytochemistry. The percentage of brain area with APP immunoreactivity within neuronal cell bodies in the impacted animals increased with time from a mean of 7.5% at 15 min to 54.5% at 2 h. Control brains showed only very small numbers of weakly APP-positive neuronal cell bodies ranging from 2 to 14% (mean 7%). Increased expression of APP mRNA was first evident in impacted hemispheres at 30 min after impact and progressively increased over time to involve neurons in all sampled regions of the brain, suggesting increased transcription of APP. In contrast, APP mRNA was undetectable in tissue from nonimpacted sheep. These data show that APP mRNA and antigen expression are sensitive early indicators of neuronal injury with widespread upregulation occurring as early as 30 min after head impact.

Peripheral heat loss: a predictor of the hypothermic response to melatonin administration in young and older women.

Core hypothermia following daytime melatonin administration typically displays significant interindividual variability. As this hypothermia has been associated with significant increases in skin temperature, the mechanism by which melatonin decreases core temperature may involve increasing peripheral heat loss. If so, the interindividual variability in this effect may reflect concomitant interindividual variability in heat loss capacity at the distal periphery. For six younger (mean +/- SEM: 23.4 +/- 0.3 years) and 10 older women (mean +/- SEM: 65.6 +/- 0.7 years), the maximum decrease in core body temperature following a 5-mg (p.o.) dose of melatonin was correlated with the capacity to lose heat. This was determined by the maximum increase in contralateral hand temperature following a mild positive thermal challenge (PTC). The regression analysis yielded a significant (p < 0.01) correlation of 0.80, suggesting that the individual magnitude of hypothermia following melatonin administration may reflect the capacity of an individual to dissipate heat at the distal periphery.

Sleeping with an electric blanket: effects on core temperature, sleep, and melatonin in young adults.

Previous studies have inferred a relationship between core temperature and sleep disruption from manipulations of core temperature such as heating prior to sleep or administration of hyperthermic substances. To examine the relationship more directly, this study aimed to produce a direct increase in core temperature during the sleep period. Following an adaptation night, 16 subjects underwent counter-balanced baseline and experimental conditions, on non-consecutive nights between 1900 and 0800h. In the experimental condition, subjects were heated between 0230h and wake up, which significantly increased mean core temperature from baseline levels between 0400 and 0700h by 0.18 +/- 0.03 degree C (mean +/- SEM, p < 0.05). This increase in core temperature was associated with a significant decrease in sleep efficiency between 0330 and 0730h of 5.5 +/- 0.9% (mean +/- SD, p < 0.05). Polysomnographic measures indicated a significant increase in the number of stage changes and the amounts of stage 0 and stage 1 sleep (p < 0.05). Other stages of sleep and the number and duration of arousals were not significantly effected by heating. There was a strong trend toward and increase in the number of arousals (p = 0.054), however, core body temperature did not increase across arousals. Also, melatonin output was not effected by heating. Taken together, these results suggest that increased nocturnal core temperature alone may disrupt sleep. Additionally, the results support evidence suggesting that the circadian regulation of the sleep/wake cycle may be mediated via core temperature.

Thermoregulatory and soporific effects of very low dose melatonin injection.

The effect of a rapid increase in circulating melatonin on body temperatures and sleepiness was investigated in eight young adults at 1000. Melatonin administered intravenously at 10- and 30-microgram doses, but not 3 microgram, resulted in elevated plasma and saliva levels consistent with endogenous levels measured in adults at night. Melatonin at 10 and 30 microgram significantly attenuated the daytime increase in rectal core temperature (P < 0.05 for both). The mean maximum rectal core temperature differences between saline and melatonin treatment were 0.11 +/- 0.03 degreesC, 0.16 +/- 0.04 degreesC, and 0.18 +/- 0.04 degreesC after the 3-, 10-, and 30-microgram melatonin doses, respectively. All three doses significantly increased hand temperature compared with saline (P < 0. 05) within 30 min. The mean maximum hand temperature differences were 0.72 +/- 0.12 degreesC (3 microgram), 0.95 +/- 0.15 degreesC (10 microgram), and 0.65 +/- 0.11 degreesC (30 microgram). Foot temperature and subjective sleepiness measures did not change at any melatonin dose. The results suggest that daytime intravenous injection of melatonin to achieve normal nocturnal levels in young adults may produce significant thermoregulatory changes without soporific effects.

Daytime melatonin and temazepam in young adult humans: equivalent effects on sleep latency and body temperatures.

1. As changes in core body temperature are generally associated with concomitant changes in sleep propensity, it is possible that the effects of hypnotic/soporific agents may be related to changes in thermoregulation. Therefore, to increase our knowledge of the mechanisms by which these agents exert their soporific effects, we compared the thermoregulatory and soporific effects of temazepam (20 mg per os (p.o.)) with those of melatonin (5 mg p.o.) when administered at 14.00 h to 20 young healthy adults (13 male, 7 female; age, 23.5 +/- 0.4 years). 2. From 08.00 to 20.30 h, subjects lay in bed, and foot and rectal (Tc) temperatures were recorded. Sleep onset latency (SOL) was measured using 20 min multiple sleep latency tests, performed hourly from 11.00 to 20.00 h, during which time heart rate was recorded. 3. Compared with placebo, both melatonin and temazepam significantly reduced Tc (-0.17 +/- 0.02 and -0.15 +/- 0.03 C, respectively) and SOL (by 4.8 +/- 1.49 and 6.5 +/- 1.62 min, respectively). Although both treatments significantly increased heat loss, only melatonin demonstrated cardiac effects. Importantly, there was a temporal relationship between minimum SOL and the maximum rate of decline in Tc for both melatonin (r = 0.48) and temazepam (r = 0.44). 4. A possible role of thermoregulation in sleep initiation is suggested by the similar temporal relationship between Tc and SOL for two different classes of soporific agents.

Visuo-motor adaptation in smokeless tobacco users.

Ten smokeless tobacco (ST) users and 11 non-smokers participated in a visuo-motor adaptation experiment in which the visual feedback of point-to-point horizontal arm movements, displayed in real-time on a computer screen, was rotated by 45 degrees counterclockwise for some trials. Visuo-motor performance between smokers and non-smokers was compared on three occasions, once after at least 8 h of tobacco abstinence (Session 1), a second time following ST intake (Session 2), and a third time 45 min after the original ST intake (Session 3). Non-smokers were tested at the same relative times as the smokers in the absence of any tobacco. Both groups performed the three conditions during each session: baseline (normal visual feedback), rotated visual feedback (45 degrees visual feedback rotation), and post-adaptation (normal visual feedback immediately following feedback rotation). Compared with non-smokers, ST users had significantly larger normalized jerk scores (a measure of movement smoothness) after ST intake during the adaptation and post-adaptation conditions in Sessions 2 and 3, but not during the baseline conditions, implying a differential effect of ST use specific to rotated visual feedback. Movement duration was also longer for smokers than for non-smokers after ST intake, but only in the post-adaptation condition. Overall the results suggest that ST use, and hence nicotine, has a detrimental effect on visuo-motor performance, particularly on movement smoothness.

Traumatic axonal injury in lambs: a model for paediatric axonal damage.

Axonal injury (AI), an important determinant of clinical outcome after traumatic head injury in adults, has been little studied in child neurotrauma. In this experimental model, 10 anaesthetised, ventilated and physiologically monitored 4-5 week old lambs sustained a temporal impact to an unconstrained head. Examination of the brain 2 hours post-impact using amyloid precursor protein immunostaining, a sensitive marker of axonal damage, revealed widespread multifocal AI in the ipsilateral and contralateral hemispheres, brain stem and cerebellum in all cases. Impact contusions (predominantly microscopic) (8 10 ), contralateral contusions (2 10 ), parenchymal haemorrhage (10 10 ) and focal subarachnoid haemorrhage (10 10 ) were also present. Unusual multifocal cerebellar granular layer necrosis occurred in all impacted lambs. The pattern of AI in these lamb brains 2 hours post-impact was similar to that found in adult sheep using the same experimental paradigm.