Introduction: Traumatic brain injury (TBI) can result in an acute coagulopathy including platelet dysfunction that can contribute to ongoing intracranial hemorrhage. Previous studies have shown ADP-induced platelet aggregation to be reduced after TBI. In addition, circulating microvesicles (MVs) are increased following TBI and have been shown to play a role in post-TBI coagulopathy and platelet function. We hypothesized that post-TBI MVs would affect platelet aggregation in a murine head injury model. Methods: Moderate TBI was performed using a weight-drop method in male C57BL6 mice. Whole blood, plasma, MVs, and MV-poor plasma were isolated from blood collected 10 minutes following TBI and were mixed separately with whole blood from uninjured mice. Platelet aggregation was measured with Multiplate impedance platelet aggregometry in response to adenosine diphosphate (ADP). The ADP P2Y 12 receptor inhibitor, R-138727, was incubated with plasma and MVs from TBI mice, and platelet inhibition was again measured.Results: Whole blood taken from 10 minute post-TBI mice demonstrated diminished ADPinduced platelet aggregation compared to sham mice. When mixed with normal donor blood, post-TBI plasma and MVs induced diminished ADP-induced platelet aggregation compared to sham plasma and sham MVs. By contrast, the addition of post-TBI MV-poor plasma to normal blood did not change ADP-induced platelet aggregation. The observed dysfunction in post-TBI ADP platelet aggregation was prevented by the pretreatment of post-TBI plasma with R-138727. Treatment of post-TBI MVs with R-138727 resulted in similar findings of improved ADP-induced platelet aggregation compared to non-treated post-TBI MVs.

Background: We investigated the influence of a botanical agent to improve sleep quality and associated measures in men and women with self-reported difficulty sleeping. Methods: 32 individuals were randomly assigned in double blind manner to ingest a botanical agent (CLOCK®, containing Rosemary [Rosmarinus officinalis] and Daylily [Hemerocallis fulva]) or a placebo over a 6-week intervention. During weeks 1 and 2, subjects ingested one serving of the assigned condition, followed by a two-week washout. During weeks 5 and 6, subjects ingested two servings of the condition. The Leeds Sleep Evaluation Questionnaire was used as an outcome measure, as were subjective measures of sleep quality, energy level, and mood. Blood samples collected pre-and post-intervention were assayed for acetylcholine (ACH), brainderived neurotrophic factor (BDNF), irisin, and melatonin. Results: No differences were noted between conditions in measures of sleep (p > 0.05). While no differences of statistical significance were noted in subjective feelings, during weeks 5 and 6 as compared to baseline, subjects assigned to the supplement noted an 8% increase in attentiveness, an 11% increase in alertness, a 12% increase in focus, a 14% increase in feeling energetic, a 12% increase in enthusiasm, a 23% increase in feeling well rested, an 11% decrease in feeling sluggish, and a 16% decrease in feeling depressed, without the same improvement observed for subjects in the placebo group. All biochemical measures were increased from pre-to post-treatment with two servings of the supplement; the largest percent increase noted for BDNF (27%) and the largest effect 1308 mood, with a significant impact on the biochemical marker, BDNF. Future studies using a larger sample size and perhaps a cross-over design may help to further clarify the impact of this dietary supplement on aspects of sleep quality, mood, and other related variables.

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D-aspartic acid (DAA), nitrate, and vitamin D 3 have received considerable attention in recent years for their potential health-enhancing properties. Using an open-label design, we evaluated the impact of a DAA/sodium nitrate/ vitamin D 3 dietary supplement on blood testosterone and nitrate/nitrite, as well as subjective indicators of health, in middle-aged men.Methods: 10 overweight or obese men (mean age: 42 years) were assigned to ingest a DAA/sodium nitrate/ vitamin D 3 supplement (either one or two servings per day) for 28 days. Blood total and free testosterone and nitrate/nitrite was measured before and after 14 and 28 days of supplementation. Subjective assessment of to health indicators (e.g., energy level, libido) was included at each collection time.Results: Total and free testosterone increased on average 5-10%, which was not of statistical significance (p>0.05). The response was highly variable; some men failed to respond to treatment, while men with relatively low basal testosterone values experienced increases exceeding 20%. Plasma nitrate/nitrite was increased approximately 6-10 fold after treatment with the supplement, with a trend noted for a time effect (p=0.07). Men reported a significantly better feeling following supplement use, as evidenced by a time effect for both vitality (p=0.02) and libido (p=0.04), with a trend noted for increased energy level (p=0.08) and mental outlook and mood (p=0.10). Conclusion:Twenty-eight days of treatment with a DAA/sodium nitrate/ vitamin D 3 dietary supplement increased blood nitrate/nitrite and improved subjective feelings of vitality and libido in middle-aged men. In selected men with low basal testosterone values, the supplement increased circulating levels of this hormone.

Attention has been given recently to herbal dietary supplements proposed to elevate testosterone and nitric oxide. This study evaluated the impact of a supplement containing Spilanthes acmella extract and Orchis latifolia extract on total blood testosterone, cortisol, and nitrate/nitrite in healthy men. Methods: Thirteen men (25.0±1.0 years) were randomly assigned (double-blind, cross-over design) to ingest a supplement (containing Spilanthes acmella extract and Orchis latifolia extract) and a placebo daily for 14 days, with a 14-day washout period between assignments. Fasting blood samples were collected on the mornings of days 1, 4, 8, and 15 and analyzed for testosterone, cortisol, and nitrate/nitrite. On day 15, subjects ingested an acute dose of the supplement or placebo and blood was collected every 30 minutes for three hours, and analyzed for testosterone. Results: No increase of significance was noted for any biochemical variable (p>0.05). However, a mean increase in testosterone from day 1 to day 15 of 29% was observed for the 13 subjects when ingesting the supplement, with a mean increase of 56% noted when only considering the 8 subjects who -responded‖ to treatment. Cortisol was increased approximately 19% when subjects ingested the supplement, compared to only 9% with the placebo. Conclusion: Two weeks of supplementation with an herbal preparation containing Spilanthes acmella extract and Orchis latifolia extract can increase testosterone in selected young men. The supplement also results in a moderate increase in cortisol. Larger scale studies are needed to further evaluate the impact of this herbal combination on testosterone in men.