Sleep habits in volunteers were assessed for 7 days before sleep deprivation using a sleep protocol to exclude interferences of the sleep rhythm. For sleep deprivation, volunteers were requested to stay awake for one night without consuming any coffee or alcohol during that night; intake of food was allowed. All LPs were done between 10:00 and 12:00 AM, and each time peripheral blood was collected. All samples revealed no pathognomonic cell counts, CSF/serum albumin ratios or oligoclonal bands.In all subjects, we measured CSF and serum levels of oleoylethanolamide along with anandamide. To quantify FAEs, CSF aliquots and serum were spiked with 25 pmol of [2 H4]-anandamide and [2 H4]-oleoylethanolamide and prepared for further analysis as described previously . Fatty acid ethanolamides were purified and quantified by isotope dilution high performance liquid chromatography/massspectrometry using a HP 1100 Series HPLC/ MS system equipped with a octadecylsilica Hypersil column . MS analyses were performed with an electro-spray ion source as previously described . Statistical analyses were performed using SPSS and R software . Because of apparent non-normality of empirical data distributions, location differences in measurements were assessed by Wilcoxon signed rank or rank sum test .Our results show a significant increase of oleoylethanolamide in human CSF after 24 h of sleep deprivation,planting table whereas the levels of the endocannabinoid anandamide remain unaffected. Interestingly, there is no indication for a functional role of anandamide in sleep induction in humans as hypothesized previously .
In rats, Murillo-Rodriguez et al. showed that anandamide increases slow wave sleep, which can be blocked by administration of the cannabinoid CB1-receptor antagonist rimonabant, indicating that the endocannabinoid system is involved in sleep regulation . Further, recent data in rats indicate diurnal variations of anandamide and oleoylethanolamide in CSF with maximum values for oleoylethanolamide during the late light phase, decreasing in the dark phase . This research was done in rats, which are awake in the lights-off period. Translating these results to the human sleep/wake cycle it may be hypothesized that the concentration of oleoylethanolamide in CSF may decrease during daytime and increase during sleep. Given these assumptions, the increase of oleoylethanolamide is not very likely the result of an accumulation, particularly with regard to the very limited extracellular life span of endocannabinoids . As in our study the second lumbar puncture was done at the same time of the day as the first one; circadian rhythms should not confound our results. Oleoylethanolamide is renowned for modulation of feeding, body weight and lipid metabolism, as its levels decrease during food deprivation and increase upon feeding . This should not be a confounder as well, as oleoylethanolamide plays its role as a local satiety hormone in the intestine , and the feeding conditions were not changed before and after sleep-withdrawal in this study. Like anandamide, oleoylethanolamide is synthesized by neurons and other cells in a stimulus-dependent manner and is rapidly eliminated by enzymatic hydrolysis. Oleoylethanolamide binds with high affinity to the PPAR-a, a nuclear receptor that is known to regulate several aspects of lipid metabolism and induces satiety by activating PPAR-a . PPAR-a is also involved in neuroprotection through activation of antioxidant and anti-inflammatory mechanisms.
It has been shown that pretreatment with the synthetic PPAR-a agonist fenofibrate is neuroprotective against cerebral injury . Furthermore, treatment with PPAR-a agonists as well as monounsaturated and polyunsaturated fatty acids induces an increase in activity of major antioxidant enzymes in the brain . The distribution of PPAR-a receptors in the human brain remains mainly conjectural. Moreno et al. have provided an overview on the distribution of PPARs in the adult rat central nervous system . They reported the highest densities of PPAR-a in the hippocampal dentate gyrus and the granular cell layer of the cerebellar cortex. Furthermore, they observed PPAR expression in previously unreported locations, such as the basal ganglia, the reticular formation, some thalamic, mesencephalic and cranial motor nuclei and the large motor neurons of the spinal cord . This widespread distribution in the CNS also includes brain areas involved in sleep regulation such as the thalamus and the reticular formation. It has been hypothesized that sleep may serve an antioxidant function by removing free radicals or oxygen reactive species produced during waking time and that restoration of antioxidant balance is a property of recovery sleep . However, these protective mechanisms, especially in the brain, are still poorly understood. Interestingly, several studies have found that sleep deprivation is a stressful condition, which is associated with the disruption of various physiological processes . Reduced glutathione levels have been found in different brain regions, such as thalamus and hypothalamus, as well as in different organs of sleepdeprived animals . Additionally, markers of generalized cell injury accompanied these decreases and decreased superoxide-dismutase activity has been shown in hippocampus and brainstem after prolonged sleep deprivation .
The activation of these components of the antioxidant defense system suggests that sleep deprivation is a stressful condition for the entire body and for the brain in particular. Several studies have shown that PPAR-a activation enhances antioxidative enzyme activities . Activation of PPAR-a in vivo causes an upregulation of the mRNA and protein levels of a number of peroxisome- and nonperoxisome-associated enzymes and structural proteins, including the antioxidant enzymes catalase, superperoxide-dismutase and mediators of the glutathione pathway. In this context, pretreatment with fenofibrate reduces cerebral infarct volume in apolipoprotein E-deficient mice. The neuroprotective effect of fenofibrate is completely absent in PPAR-a-deficient mice, suggesting that PPAR-a activation is involved as a protective mechanism against cerebral injury . We failed to confirm the initially hypothesized increase of anandamide after sleep deprivation questioning its proposed role in sleep induction in humans . This is in line with findings on cannabinoid CB1- receptor gene expression, which is suggested to be modulated by sleep. While sleep rebound significantly increased CB1-receptor protein and decreased respective mRNA, no effects were found following sleep deprivation . Interestingly, according to the product information, the CB1-receptor antagonist rimonabant induces sleep disturbances frequently but also sedation in clinical trials. Several shortcomings might have influenced our somehow preliminary data; first, a randomized, balanced crossover design would have been the ideal design for the trial. Second, EEG recordings might have provided more detailed information on sleep quality instead of an actigraphy for control of sleep deprivation only.More high school students smoked little cigars and cigarillos than cigarettes in 33 US states in 2015. Concern is growing about co-use of tobacco and marijuana among youth, particularly among African-American youth.In a 2015 survey, for example, one in four Florida high school students reported ever using cigars or cigar wraps to smoke marijuana. One colloquial term for this is a “blunt.” Adolescent cigar smokers were almost ten times more likely than adults to report that their usual brand offers a flavored variety. Since the US ban on flavored cigarettes ,cannabis indoor grow system the number of unique LCC flavors more than doubled. Anticipating further regulation, the industry increasingly markets flavored LCCs with sensory and other descriptors that are not recognizable tastes. For example, after New York City prohibited the sale of flavored cigars, blueberry and strawberry cigarillos were marketed as blue and pink, but contained the same flavor ingredients as prohibited products. Among the proliferation of such “concept” flavors , anecdotal evidence suggests that references to marijuana are evident.Cigar marketing includes the colloquial term, “blunt”, in brand names and product labels . Other marketing techniques imply that some brands of cigarillos make it easier for users to replace the contents with marijuana.For example, the image of a zipper on the packaging for Splitarillos and claims about “EZ roll” suggest that products are easily manipulated for making blunts.
We use the term “marijuana co-marketing” to refer to such tobacco industry marketing that may promote dual use of tobacco and marijuana and concurrent use . In addition to flavoring, low prices for LCCs also likely increase their appeal to youth. In California, 74% of licensed tobacco retailers sold cigarillos for less than $1 in 2013. Before Boston regulated cigar pack size and price in 2012, the median price for a popular brand of grape-flavored cigars was $1.19. In 2012, 78% of US tobacco retailers sold single cigarillos, which suggests that the problem of cheap, combustible tobacco is widespread.Additionally, the magnitude of the problem is worse in some neighborhoods than others. Popular brands of flavored cigarillos cost significantly less in Washington DC block groups with a higher proportion of African Americans and in California census tracts with lower median household income.For the first time, this study examines neighborhood variation in the maximum pack size of cigarillos priced at $1 or less and assesses the prevalence of marijuana co-marketing in the retail environment for tobacco. School neighborhoods are the focus of this research because 78% of USA teens attend school within walking distance of a tobacco retailer.In addition, emerging research suggests that adolescents’ exposure to retail marketing is associated with greater curiosity about smoking cigars15 and higher odds of ever smoking blunts.In California, 79% of licensed tobacco retailers near public schools sold LCCs and approximately 6 in 10 of these LCC retailers sold cigar products labeled as blunts or blunt wraps or sold cigar products with a marijuana-related flavor descriptor. A greater presence of marijuana co-marketing in neighborhoods with a higher proportion of school-age youth and lower median household income raises concerns about how industry marketing tactics may contribute to disparities in LCC use. The study results also suggest that $1 buys significantly more cigarillos in California school neighborhoods with lower median household income. Policies to establish minimum pack sizes and prices could reduce the widespread availability of cheap cigar products and address disparities in disadvantaged areas.After Boston’s 2012 cigar regulation, the mean price for a grape-flavored cigar was $1.35 higher than in comparison communities. 12 The industry circumvented sales restrictions in some cities by marketing even larger packs of cigarillos at the same low price,and the industry’s tipping point on supersized cigarillo packs for less than $1 is not yet known. The retail availability of 5- and 6-packs of LCCs for less than $1 observed near California schools underscores policy recommendations to establish minimum prices for multipacks .A novel measure of marijuana co-marketing and a representative sample of retailers near schools are strengths of the current study. A limitation is that the study assessed the presence of marijuana co-marketing, but not the quantity. The protocol likely underestimates the prevalence of marijuana co-marketing near schools because we lacked a comprehensive list of LCC brands and flavor varieties. Indeed, state and local tobacco control policy research and enforcement would be greatly enhanced by access to a comprehensive list of tobacco products from the US Food and Drug Administration, including product name, category, identification number and flavor. Both a routinely updated list and product repository would be useful for tobacco control research, particularly for further identifying how packaging and product design reference marijuana use. This first assessment of marijuana co-marketing focused on brand and flavor names because of their appeal to youth. However, the narrow focus is a limitation that also likely underestimates the prevalence of marijuana co-marketing. Other elements of packaging and product design should be considered in future assessments. Examples are pack imagery that refers to blunt making, such as the zipper on Splitarillos, as well as re-sealable packaging for cigarillos and blunt wraps, which is convenient for tobacco users who want to store marijuana. Coding for brands that are perforated to facilitate blunt making and marketing that refers to “EZ roll” should also be considered. Future research could assess marijuana co-marketing across a larger scope of tobacco/nicotine products. The same devices can be used for vaping both nicotine and marijuana. Advertising for vaping products also features compatibility with “herbs” and otherwise associates nicotine with words or images that refer to marijuana . Conducted before California legalized recreational marijuana use, the current study represents a baseline for understanding how retail marketing responds to a policy environment where restrictions on marijuana and tobacco are changing, albeit in opposite directions.The prevalence of marijuana co-marketing near schools makes it imperative to understand how tobacco marketing capitalizes on the appeal of marijuana to youth and other priority populations.