Other elements of packaging and product design should be considered in future assessments

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 , 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,grow trays 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 cigars 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.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.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.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. How drying marijuana co-marketing contributes to dual and concurrent use of marijuana and tobacco warrants study, particularly for youth and young adults. In previous research, the prevalence of adult marijuana use in 50 California cities was positively correlated with the retail availability of blunts.

Whether this is correlated with blunt use by adolescents is not yet known. Consumer perception studies are necessary to assess whether marijuana co-marketing increases the appeal of cigar smoking or contributes to false beliefs about product ingredients. Research is also needed to understand how the tobacco industry exploits opportunities for marijuana co-marketing in response to policies that restrict sales of flavored tobacco products and to policies that legalize recreational marijuana use. Such assessments are essential to understand young people’s use patterns and to inform current policy concerns about how expanding retail environments for recreational marijuana will impact tobacco marketing and use.Despite their similar chemical structures, anandamide and 2-arachidonylglycerol are produced through distinct biochemical pathways. Formation of anandamide may result from hydrolysis of the phospholipid precursor Narachidonyl phosphatidylethanolamine, catalyzed by a Ž phosphodiesterase such as phospholipase D Di Marzo et . al., 1994; Cadas et al., 1997; Schmid, 2000 . 2-Arachidonylglycerol, on the other hand, may be produced by cleavage of 1,2-diacylglycerol generated by phospholipase Ž C acting on phosphatidylinositol bisphosphate Gammon et . al., 1989; Stella et al., 1997 , although the participation of Ž alternative pathways cannot be excluded Piomelli et al., . 2000 . The existence of different enzymatic routes for the formation of anandamide and 2-arachidonylglycerol suggests that under certain circumstances, these two endocannabinoid substances might operate independently of each other. This possibility is supported by two findings. In hippocampal slices, stimulation of glutamate-releasing fibers in the Schaffer collaterals increases the levels of Ž 2-arachidonylglycerol, but not those of anandamide Stella . et al., 1997 . Conversely, in vivo microdialysis experiments show that activation of striatal D -family dopamine 2 receptors enhances release of anandamide, but not of 2- Ž . arachidonylglycerol Giuffrida et al., 1999 . It is unclear, however, whether this discrepancy reflects different experimental conditions, regional segregation of the phospholipase C and phospholipase D pathways, or receptoractivated mechanisms linked to the generation of specific endocannabinoid lipids. To examine these different possibilities, in the present study we have investigated the receptor mechanisms underlying the formation of anandamide and 2-arachidonylglycerol in primary cultures of rat cortical neurons. Our findings indicate that the excitatory neurotransmitter glutamate triggers 2-arachidonylglycerol biosynthesis by allowing external Ca2q to enter cortical neurons through activated NMDA receptor channels. In these cells, 2-arachidonylglycerol formation is likely to be mediated by the phospholipase Crdiacylglycerol lipase pathway, because selective drug inhibitors of these enzyme activities prevent the accumulation of 2-arachidonylglycerol elicited 2q Ž . by a Ca ionophore Stella et al., 1997 . This conclusion is also in agreement with previous work showing that NMDA receptor activation stimulates phospholipase C and Ž diacylglycerol lipase activities in cultured neurons Nico- . letti et al., 1986; Farooqui et al., 1993 . Noteworthy, we found that activation of acetylcholine receptors with carbachol had little or no effect, per se, on 2-arachidonylglycerol levels.

This result, at variance with the ability of cholinergic agonists to stimulate phospholipase C activity in many tissues and to cause 2-arachidonylglycerol accu- Ž . mulation in vascular endothelium Mechoulam et al., 1998 , underscores the differences in 2-arachidonylglycerol biosynthesis between neurons and other non-neuronal cell types. The functional role of 2-arachidonylglycerol formation in NMDA receptor signaling is unclear at present. However, experiments with superfused hippocampal slices suggest that 2-arachidonylglycerol may act as a localized Ž feedback signal within the hippocampus Stella et al., . 1997 . Electrical stimulation of the Schaffer’s collaterals, a glutamatergic fiber tract that projects from CA3 to CA1 neurons, produced a marked increase in 2-arachidonylglycerol accumulation in the slices, which was prevented by the Naq channel blocker tetrodotoxin or by removal of 2q Ž . external Ca Stella et al., 1997 . In the same preparation, exogenous 2-arachidonylglycerol inhibited the induction of long-term potentiation at CA3–CA1 synapses by activating local cannabinoid CB receptors, whereas it had no 1 Ž . effect on basal synaptic transmission Stella et al., 1997 . The possible role of 2-arachidonylglycerol as a negative feedback regulator of N-methyl-D-aspartate-mediated responses, suggested by these results, is reinforced by the ability of cannabinoid drugs to reduce glutamate transmis- Ž . sion Shen et al., 1996; Shen and Thayer, 1999 , inhibit Ž long-term potentiation Collins et al., 1994; Terranova et . al., 1995; Misner and Sullivan, 1999 , and alleviate gluta- Ž . mate-induced neurotoxicity Nagayama et al., 1999 .In the present experiments, the simultaneous application of glutamate and carbachol, but not of either agent alone, caused a marked stimulation of anandamide biosynthesis in cortical neurons. Pharmacological experiments suggest that this synergistic effect may result from the coactivation of NMDA and a7 nicotinic receptors, and may depend both on membrane depolarization and on mobilization of Ca2q ions from intracellular stores. Though necessary for the response, membrane depolarization was insufficient per se to initiate anandamide biosynthesis.Thus, fatty acid ethanolamide biosynthesis in the former cultures might result, not from a direct effect of membrane depolarization, but from the action of neurotransmitters released in the extracellular medium. A similar indirect mechanism may be responsible for the release of anandamide induced in vivo by a depolarizing concentration of KCl, administered in dorsal striatum by Ž . reverse dialysis Giuffrida et al., 1999 . Receptor-dependent anandamide formation may be distinguished pharmacologically from that of other fatty acid ethanolamides that do not activate cannabinoid receptors. Indeed, though biosynthesis of all fatty acid ethanolamides is contingent on NMDA receptor occupation, anandamide formation requires the coactivation of NMDA and a7 nicotinic receptors, while oleylethanolamide and palmitylethanolamide formation requires the coactivation of NMDA and muscarinic receptors. Thus, glutamate and acetylcholine may elicit the biosynthesis of different fatty acid ethanolamides, depending on the complement of acetylcholine receptors expressed in their target neurons. The finding that palmitylethanolamide and oleylethanolamide may be produced by dissimilar molecular mechanisms and may exert biological effects that are not medi- Ž ated by cannabinoid receptors Calignano et al., 1998; . Jaggar et al., 1998 raises the possibility that they might serve signaling functions independent from those of anandamide. Testing this possibility will require a thorough investigation of the pharmacological effects of palmitylethanolamide and oleylethanolamide in neurons, as well as the identification of the putative cellular targets that may mediate these effects. In conclusion, biosynthesis of anandamide and 2- arachidonylglycerol in cortical neurons may be triggered by activation of membrane receptors for the neurotransmitters, glutamate and acetylcholine. This suggests that a key function of the endocannabinoid system in the brain may be to modulate the effects of primary neurotransmitters by a localized feedback action.