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mRNA encoding for the CB2 cannabinoid receptor is expressed by many subsets of human peripheral blood leukocytes (PBL), but little is known about the resulting protein expression and function. Employing clones from the A549 and 293T cell lines that were constructed to express both full-length human CB2 and GFP, we developed a flow cytometry assay for characterizing CB2 protein expression. A monoclonal antibody directed against human CB2 selectively stained the surface of transduced but not parental cell lines. When cells were fixed and permeabilized, imaging flow cytometry identified large stores of intracellular protein. Total cellular staining for CB2 corresponded closely with the level of GFP expression. When exposed to Δ-9-tetrahydrocannabinol, CB2-expressing cells internalized cell surface CB2 receptors in a time- and dose-dependent manner. Applying these approaches to human PBL, CB2 protein was identified on the surface of human B cells but not on T cells or monocytes. In contrast, when PBL were fixed and permeabilized, intracellular CB2 expression was readily detected in all three subsets by both conventional and imaging flow cytometry. Similar to the protein expression pattern observed in fixed and permeabilized PBL, purified B cells, T cells, and monocytes expressed relatively equal levels of CB2 mRNA by quantitative real-time RT-PCR. Our findings confirm that human PBL express CB2 protein but that its distribution is predominantly intracellular with only B cells expressing CB2 protein at the extracellular membrane. The differential role of intracellular and extracellular CB2 receptors in mediating ligand signaling and immune function remains to be determined.

Oxidative stress has been proposed as a central causal mechanism underlying the life‐history trade‐off between current and future reproduction and survival in wild animals. While mixed evidence suggests that maternal oxidative stress may act both as a constraint and a cost to reproduction, some studies have reported a lack of association between reproduction and maternal oxidative stress. The oxidative shielding hypothesis offers an alternative explanation, suggesting that mothers may pre‐emptively mitigate the oxidative costs of reproduction by increasing antioxidant defences prior to reproduction. We tested the oxidative constraint, cost and shielding hypotheses using a longitudinal field study of oxidative stress levels in a species that breeds using daily energy income, the Columbian ground squirrel (Urocitellus columbianus). Elevated maternal oxidative damage prior to reproduction was associated with higher maternal investment in litter mass at birth, but not at weaning. Breeding females increased their antioxidant capacity and decreased their oxidative damage from birth to lactation, compared to non‐breeding females measured at the same time periods. However, lower maternal oxidative stress during lactation was not associated with higher offspring survival or mass growth over this period. Our results provide little evidence for maternal oxidative stress acting as a constraint on, or cost to, reproduction in Columbian ground squirrels, but partially support the idea that oxidative shielding occurred to buffer potential oxidative costs of reproduction. A http://onlinelibrary.wiley.com/doi/10.1111/1365-2435.13032/suppinfo is available for this article.

ABSTRACT. Objective: Empirical knowledge is lacking about AlAnon Family Groups (Al-Anon), the most widely used form of help by people concerned about another's drinking, partly because conducting research on 12-step groups is challenging. Our purpose was to describe a new method of obtaining survey data from 12-step group attendees and to examine infl uences on initial Al-Anon attendance and attendees' recent life contexts and functioning. Method: Al-Anon's World Service Offi ce sent a mailing to a random sample of groups, which subsequently yielded surveys from newcomers (n = 359) and stable members (n = 264). Results: Reasons for groups' nonparticipation included having infrequent newcomers and the study being seen as either contrary to the 12 Traditions or too uncomfortable for newcomers. Main concerns prompting initial Al-Anon attendance were problems with overall quality of life and with the Al-Anon trigger (a signifi cant drinking individual), and being stressed and angry. Goals for Al-Anon attendance were related to the following concerns: better quality of life, fewer trigger-related problems, and less stress. Members reported better functioning in some of these domains (quality of life, relationship with the trigger) but did not differ from newcomers on physical and psychological health. Newcomers were more likely to have recently drunk alcohol and to have obtained treatment for their own substance misuse problems. Conclusions: This method of collecting data from 12-step group attendees yielded valid data and also was seen by many in Al-Anon as consistent with the Traditions. Both newcomers and members had aimed to improve their overall quality of life and well-being through Al-Anon, and, indeed, members were more satisfi ed with their quality of life than were newcomers. (J. Stud. Alcohol Drugs, 74, 965-976, 2013)

Background and Objectives Despite Al-Anon’s widespread availability and use, knowledge is lacking about the drinkers in attendees’ lives. We filled this gap by describing and comparing Al-Anon newcomers’ and members’ reports about their “main drinker” (main person prompting initial attendance). Methods Al-Anon’s World Service Office mailed a random sample of groups, yielding completed surveys from newcomers (N = 362) and stable members (N = 265). Results Newcomers’ and members’ drinkers generally were comparable. They had known their drinker for an average of 22 years and been concerned about his or her’s drinking for 9 years; about 50% had daily contact with the drinker. Most reported negative relationship aspects (drinker gets on your nerves; you disagree about important things). Newcomers had more concern about the drinker’s alcohol use than members did, and were more likely to report their drinkers’ driving under the influence. Drinkers’ most frequent problem due to drinking was family arguments, and most common source of help was 12-step groups, with lower rates among drinkers of newcomers. Concerns spurring initial Al-Anon attendance were the drinker’s poor quality of life, relationships, and psychological status; goals for initial attendance reflected these concerns. Discussion and Conclusions The drinker’s alcohol use was of less concern in prompting initial Al-Anon attendance, and, accordingly, the drinker’s reduced drinking was a less frequently endorsed goal of attendance. Scientific Significance Family treatments for substance use problems might expand interventions and outcome domains beyond abstinence and relationship satisfaction to include the drinker’s quality of life and psychological symptoms and in turn relieve concerns of family members.