Genome engineering with targetable nucleases.

Current technology enables the production of highly specific genome modifications with excellent efficiency and specificity. Key to this capability are targetable DNA cleavage reagents and cellular DNA repair pathways. The break made by these reagents can produce localized sequence changes through inaccurate nonhomologous end joining (NHEJ), often leading to gene inactivation. Alternatively, user-provided DNA can be used as a template for repair by homologous recombination (HR), leading to the introduction of desired sequence changes. This review describes three classes of targetable cleavage reagents: zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas RNA-guided nucleases (RGNs). As a group, these reagents have been successfully used to modify genomic sequences in a wide variety of cells and organisms, including humans. This review discusses the properties, advantages, and limitations of each system, as well as the specific considerations required for their use in different biological systems.

Estimating the causal effect of filter ventilation levels in cigarettes on past 30-day smoking.

INTRODUCTION: Cigarettes with higher levels of filter ventilation are misperceived as less harmful and may be more appealing to consumers. Setting limits on filter ventilation has been considered as a policy, but a better understanding of any potential unintended consequences is needed. METHODS: Filter ventilation (0.2-61.1%) measured for 114 subbrands was merged with Wave 1 (2012-2013) of the Population Assessment of Tobacco Use and Health (PATH) data, restricted to adults 25+ years of age who smoked daily, and examined by quartiles. Inverse probability of exposure weights were used to estimate the causal effect of filter ventilation on past-30 day smoking at subsequent waves while accounting for potential confounders including demographics, menthol, heaviness of smoking and past quit attempts. RESULTS: Compared to those in the 1st (lowest) quartile of filter ventilation, those in the 2nd, 3rd and 4th quartiles had 1.02 (95% confidence interval: 0.57, 1.82), 0.86 (0.42, 1.73) and 1.52 (0.90, 2.56) times the odds of no past 30-day smoking at Wave 2 (approximately 1 year later, p=0.163), and 1.28 (0.80, 2.07), 1.11 (0.67, 1.83) and 1.65 (1.01, 1.24) times the odds of no past 30-day smoking at Wave 4 (3 years later, p = 0.238). CONCLUSIONS: This observational study found no strong evidence of a causal effect of filter ventilation on past 30-day smoking at approximately 1 and 3 years follow-up. However, our effect size estimates were not precise and thus an increase in the ability to quit smoking due to higher filter ventilation levels cannot be ruled out. IMPLICATIONS: Setting a maximum limit on filter ventilation (FV) in cigarettes could address the misperception that highly ventilated cigarettes are less harmful and the link between FV and lung adenocarcinoma. It is important to understand whether such a policy would have unintended consequences on longer-term smoking behavior. We found no strong evidence that FV affects past 30-day smoking 1-3 years later, but could not rule out the possibility that higher FV increases cessation rates. If future studies confirm these epidemiologic findings, this could mean that setting a limit on FV would not lead to reductions in the ability to quit smoking.

American Indian Perspectives on Culturally Aligning a Digital Smoking Cessation Resource.

INTRODUCTION: Innovative smoking cessation approaches that overcome barriers such as traveling to program site or that require the staff and infrastructure for sustaining are likely needed to improve smoking quit rates among American Indian (AI) peoples in the United States. In this study, qualitative methods identified recommendations from AI peoples to guide alignment of an evidence-based smoking cessation smartphone app (i.e., QuitGuide) to the culture and needs of AI persons. METHODS: Semi-structured interviews were conducted with AI adults who smoke (n = 40) and with public health professionals (n = 6). Questions included: "The app asks if something triggered you to slip and lists several options. What options were you expecting to see on this list?" as well as how to make the app more engaging such as "What would make the app more helpful for AI peoples, like you, who want to quit smoking?." Constant comparative techniques were used to develop codes and themes. RESULTS: Loss, grief, and not accessing traditional tobacco were put forward as smoking triggers to be addressed in the app. Features that help users connect with and learn about AI cultures and promote healing, such as encouraging traditional tobacco use, being in community, embracing Native spirituality, and participating in cultural crafting were recommended. Some noted the need to motivate AI peoples to think about legacy and ability to care for younger generations and Indigenizing the app with Native imagery. CONCLUSIONS: Themes pointed towards promotion of strengths-based factors, such as healing, cultural connectedness and traditional tobacco use, in the app. IMPLICATIONS: Results will be used to culturally align a smartphone app for smoking cessation among AI peoples and may be insightful for other tribal, federal, and state public health efforts aimed at advancing health equity for AI peoples.

Menthol and filter ventilation in cigarettes: prevalence estimates and relationships with harm perception and smoking exposure.

INTRODUCTION: Menthol and filter ventilation (FV) contribute to cigarette appeal. This observational study examines the US prevalence of menthol versus non-menthol cigarette use by FV and how harm perceptions, cigarettes per day and biomarkers of exposure vary. METHODS: Population Assessment of Tobacco and Health Study (2013-2014) was merged with FV levels of cigarettes and restricted to daily smoking adults who had a usual cigarette variety and did not regularly use other tobacco (N=1614). Weighted descriptive statistics identified the prevalence of menthol and non-menthol use by low (0.02%-10.04%), moderate (10.05%-23.40%), high (23.41%-28.12%) and very high FV (28.13%-61.10%). Weighted linear regression was used to examine differences in outcomes by menthol/FV adjusted for potential confounders. RESULTS: The prevalence of a usual brand that was non-menthol, low FV was the lowest at 2.91%. Using non-menthol cigarettes with high and very high FV (≥23.4%) vs low FV (≤10.04%) was associated with a greater likeliness of misperceiving one's cigarette variety to be less harmful than other varieties (p values<0.05). Total nicotine equivalent, biomarker for nicotine exposure, was elevated (p values<0.05) among three non-menthol groups (low, moderate and very high FV) compared with two menthol groups (moderate, very high FV). CONCLUSION: The well-documented harm misperception linked to higher FV is more apparent in those using non-menthol than menthol cigarettes. Increased exposures were observed among some non-menthol cigarette users compared with some menthol cigarette users. These results should by no means delay a menthol ban but rather motivate concerted public health efforts to accompany the menthol ban to maximise smoking cessation.

Pilot randomised controlled trial of a culturally aligned smoking cessation app for American Indian persons.

OBJECTIVE: To pilot test QuitGuide for Natives, a culturally aligned version of the National Cancer Institute's QuitGuide smartphone app for smoking cessation. METHODS: This randomised controlled trial was conducted remotely during 2022-2023. American Indian adults who smoked and resided in the Midwest (n=115) were randomised to QuitGuide for Natives or the general audience QuitGuide smartphone-based intervention. Group differences in feasibility (times the app was initiated), usability, acceptability ('How likely would you be to recommend the app to a friend?'), fit of app with culture and preliminary efficacy (24-hour quit attempts, cotinine-confirmed self-reported 7-day abstinence) outcomes were examined. RESULTS: QuitGuide for Natives versus the general audience QuitGuide did not differ in the number of times the app was opened (adjusted incidence rate ratio 0.94 (95% CI 0.63 to 1.40); p=0.743) nor in usability score (adjusted mean difference (aMD) 0.73 (95% CI: -5.00 to 6.46); p=0.801) or likeliness of recommending the app to a friend (aMD 0.62 (95% CI -0.02 to 1.27); p=0.058). Differences were observed for all cultural fit outcomes such as 'The app fits my American Indian culture (aMD 0.75 (95% CI 0.35 to 1.16); p<0.001). QuitGuide for Natives versus the general audience QuitGuide resulted in an average of 6.6 vs 5.1 24-hour quit attempts (p=0.349) and cotinine-confirmed 7-day abstinence was achieved by 6.9% vs 3.5% (p=0.679). CONCLUSIONS: Acceptability, cultural fit and preliminary efficacy findings are encouraging and will inform future, larger-scale evaluation of culturally aligned digital smoking cessation resources for American Indian adults.

Substance Use Screening in Geriatric Primary Care: Cultural Issues and Alcohol Consumption in the Deep South.

OBJECTIVES: To investigate indicators of potentially hazardous alcohol use among older adults living in a region with high substance use stigma. METHODS: Patients at a university-affiliated geriatrics clinic in the Deep South of theUS completed behavioral health screenings including self-reported alcohol use, symptoms of depression or anxiety, and cognitive functioning between 2018 and 2022. RESULTS: Participants (N = 278) averaged 76.04 years of age (SD = 9.25), were predominantly female (70.9%), and non-Hispanic white (84.5%), with an averageof 6.08 comorbid diagnoses (SD = 2.86). Race/ethnicity, age, and symptoms of anxiety were associated with alcohol use and hazardous alcohol use, with non-Hispanic whites, younger individuals, and those with more anxiety symptoms reporting more alcohol use. Notably, alcohol use and hazardous alcohol use were associated with cognitive functioning in the dementia range. CONCLUSION: Self-reported alcohol use is low in geriatric primary care in the Deep South, US, differs by race/ethnicity, and is predictive of cognitive impairment when alcohol use is hazardous. Issues of trust and stigma may play a role in self-report ofstigmatized behaviors. CLINICAL IMPLICATIONS: Self-reported alcohol intake must be considered within the cultural context of regional stigma. Recommendations to address this are provided.

Smoking Quit Attempts and Associated Factors Among Rural Adults Who Smoke Daily in the US.

INTRODUCTION: Rural American communities are heavily affected by tobacco-related health disparities. This study aims to evaluate the prevalence of quit attempts (QA) and factors that promote or impede QA among rural adults who smoke daily. METHODS: Data from Wave 5 of the Population Assessment of Tobacco and Health study were analyzed. Multivariable logistic regression was used to identify factors associated with quit attempt. Backwards selection was used to identify variables included in the final model with statistical significance set at p <0.05. RESULTS: Among 1,610 rural adults who smoked daily, the prevalence of a quit attempt in the past 12 months was 25.6% (95% CI: 23.2, 28.2). Factors associated with greater QA odds: having greater education (aOR = 1.35, 95% CI: 1.03, 1.77), e-cigarette use (aOR=1.35 95% CI: 1.03, 1.80), disapproval of smoking from friends/family (aOR= 1.46, 95% CI: 1.09-1.94), greater frequency of thinking about the harm of tobacco (aOR = 1.48, 95% CI: 1.28, 1.71), fair/poor physical health (aOR=1.31, 95% CI: 1.00, 1.70), and being advised to quit by a doctor (aOR =1.63 , 95% CI:1.25, 2.13). Smokeless tobacco use (aOR = 0.67, 95% CI: 0.47, 0.96) and greater cigarettes per day (aOR = 0.67, 95% CI: 0.47, 0.96) were associated with QA lower odds. CONCLUSION: Only 1 in 4 rural adults who smoke made a past year quit attempt. Interventions that promote provider advice to quit smoking, tobacco health harms, and normative beliefs may increase quit attempts in rural communities. IMPLICATIONS: Along with higher smoking rates and lower quitting attempts, rural communities face limited access to programs, medication, and health care professionals as tools to help them quit smoking. Public Health initiatives should focus on developing cultural sensitivity training targeting health care professionals to advise patients to quit smoking and the role of multiple tobacco products use. Furthermore, given the low rates of smoking quitting attempts future mixed methods research is needed to inform policies and interventions targeted at eliminating tobacco-related health disparities.

Exploring Potential for a Personalized Medicine Approach to Smoking Cessation With an American Indian Tribe.

INTRODUCTION: A potential precision medicine approach to smoking cessation is tailoring pharmacotherapy to a biomarker known as the nicotine metabolite ratio (NMR). Little is known about the potential impact and acceptability of this approach for American Indian (AI) persons. AIMS AND METHODS: Tribal-academic collaboration was formed and during 2019-2020 AI adults who smoke(N = 54) were recruited to (1) examine correlations between NMR, dependence, and smoking exposure; (2) assess the extent to which pharmacotherapy preference aligned with NMR-informed recommendations; (3) explore acceptability of NMR-informed pharmacotherapy selection. Participants provided samples for assessment of salivary NMR and urinary total nicotine equivalents (TNE) and completed a questionnaire that assessed cigarettes per day (CPD), Fagerstrom Test for Cigarette Dependence (FTCD), pharmacotherapy preference, and perceptions of NMR-informed pharmacotherapy selection. RESULTS: Significant positive correlations were observed between NMR and FTCD (r = 0.29;p = .0383) and its abbreviated version Heaviness of Smoking Index (HIS) (r = 0.28;p =.0426). Post-hoc analyses suggest that relationships between dependence and NMR were driven by time to first cigarette. Nonsignificant, but directionally consistent, relationships were observed between NMR and CPD (r = 0.21; p =0.1436) and TNE (r = 0.24;p = .2906). Most participants preferred nicotine replacement therapy (71%) over varenicline (29%) and preference for pharmacotherapy matched NMR-based recommendations in 54% of participants. NMR-informed pharmacotherapy selection was supported by 62% of participants. CONCLUSION: In a sample of AI adults who smoke, NMR was related to cigarette dependence and about one-half of participants' pharmacotherapy preference matched their NMR-informed recommendation. There was lower acceptability of NMR-informed approach in this sample of AI adults than prior studies among white or black/African American people who smoke. IMPLICATIONS: Relationships between NMR, dependence, and self-preference for pharmacotherapy suggest that NMR-informed pharmacotherapy selection may have potential for enhancing smoking quitting success in this Tribe. Lower acceptability of NMR-informed pharmacotherapy in this Tribe suggests that this approach may not be equitably utilized. Future work could include identifying community-driven solutions to mitigate precision medicine concerns.

Older age is associated with greater misperception of the relative health risk of e-cigarettes and cigarettes among US adults who smoke.

INTRODUCTION: The prevalence of cigarette smoking among adults aged ≥55 has remained stagnant over the past decade. National data modelling suggests no reduction in cigarette smoking prevalence attributable to e-cigarette use in the USA among people aged ≥45. Misperceptions about the absolute risks (ie, cigarettes are not harmful) and relative risks (ie, e-cigarettes are more harmful than cigarettes) of tobacco products may contribute to sustained smoking prevalence and hesitancy to switch from cigarettes to e-cigarettes among older adults. METHODS: Participants reported cigarette use (n=8072) at Wave 5 (2018-2019) of the Population Assessment of Tobacco and Health Study. Weighted multivariable logistic regressions included six age categories (independent variable) and cigarette and e-cigarette risk perceptions (outcomes). Additional models assessed the associations between dichotomous age (≥55 vs 18-54), risk perceptions and an interaction term (independent variables) with past 12-month quit attempts and past-month e-cigarette use (outcomes). RESULTS: Adults aged ≥65 were less likely than adults aged 18-24 to rate cigarettes as very/extremely harmful (p<0.05). Odds of rating e-cigarettes as more harmful than cigarettes among adults aged 55-64 and ≥65 were 1.71 (p<0.001) and 1.43 (p=0.024) greater than for adults aged 18-24. This misperception was negatively associated with past-month e-cigarette use and was stronger among adults aged ≥55 (p<0.001) than adults aged <55 (p<0.001). DISCUSSION: Adults aged ≥55 are more likely to have misperceptions about the absolute and relative risks of tobacco products, which may contribute to continued smoking. Health communications targeting this age group could modify beliefs about the perceived harms of tobacco products.

Switching to cigarette brand variants with different filter ventilation levels: a descriptive analysis.

BACKGROUND: Regulation of filter ventilation (FV) has been proposed to reduce misperceptions that ventilation reduces the health risks of smoking. We describe smoking behaviour and exposure after switching to a cigarette brand variant (CBV) with a different FV level. METHODS: Wave 1 (2013-2014) of the Population Assessment of Tobacco Use and Health Study was merged with FV levels of participants' CBV and restricted to adults with a usual CBV, smoked daily and included in wave 4 (2016-2017; n=371). Generalised estimation equations method modelled changes in FV and cigarettes per day (CPD), quit interest, total nicotine equivalents (TNE) and total NNAL (biomarker of a tobacco-specific carcinogen). FV change was defined as a change in CBV resulting in a ≥20% increase or decrease in FV. Secondary analyses used FV change based on an increase from <5% to >10% or a decrease from >10% to <5%. RESULTS: A non-significant pattern indicating an increase of 0.97 and 0.49 CPD was observed among those who switched to a CBV and increased FV by ≥20% and from <5% to >10%, respectively. A non-significant pattern indicating a decrease of 1.31 and 1.97 CPD was observed among those who decreased FV by ≥20% and from >10% to <5%, respectively. Changes in quit interest and biomarkers were also non-significant with one exception: greater reduction in TNE among those who decreased from >10% to <5% FV versus no change (-8.51 vs -0.25 nmol/mg creatinine; p=0.0447). CONCLUSIONS: Switching to CBV with lower FV does not appear to increase exposure and may even reduce exposure for some. Additional investigations are recommended to confirm these descriptive findings.

Risk perceptions and continued smoking as a function of cigarette filter ventilation level among US youth and young adults who smoke.

BACKGROUND: While evidence demonstrates that the industry's marketing of cigarettes with higher filter ventilation (FV) misleads adults about their health risks, there is no research on the relationships between FV, risk perceptions and smoking trajectories among youth (ages 12-17) and young adults (ages 18-24). METHODS: Data on FV levels of major US cigarette brands/sub-brands were merged with the Population Assessment of Tobacco and Health Study to examine whether FV level in cigarettes used by wave 1 youth/young adults (n=1970) predicted continued smoking at waves 2-4, and whether those relationships were mediated by perceived risk of their cigarette brand. FV was modelled based on tertiles (0.2%-11.8%, low; 11.9%-23.2%, moderate; 23.3%-61.1%, high) to predict daily smoking, past 30-day smoking and change in number of days smoking at successive waves. RESULTS: The odds of perceiving one's brand as less harmful than other cigarette brands was 2.21 times higher in the high versus low FV group (p=0.0146). Relationships between FV and smoking outcomes at successive waves were non-significant (all p>0.05). CONCLUSION: Youth and young adults who use higher FV cigarettes perceived their brand as less harmful compared with other brands. However, level of FV was not associated with continued smoking.

Practical guidance on modeling choices for the virtual twins method.

Individuals can vary drastically in their response to the same treatment, and this heterogeneity has driven the push for more personalized medicine. Accurate and interpretable methods to identify subgroups that respond to the treatment differently from the population average are necessary to achieving this goal. The Virtual Twins (VT) method is a highly cited and implemented method for subgroup identification because of its intuitive framework. However, since its initial publication, many researchers still rely heavily on the authors' initial modeling suggestions without examining newer and more powerful alternatives. This leaves much of the potential of the method untapped. We comprehensively evaluate the performance of VT with different combinations of methods in each of its component steps, under a collection of linear and nonlinear problem settings. Our simulations show that the method choice for Step 1 of VT, in which dense models with high predictive performance are fit for the potential outcomes, is highly influential in the overall accuracy of the method, and Superlearner is a promising choice. We illustrate our findings by using VT to identify subgroups with heterogeneous treatment effects in a randomized, double-blind trial of very low nicotine content cigarettes.

Psychometric Properties of the Acceptance and Action Questionnaire-II (AAQ-II) in Older Adult Primary Care Patients.

OBJECTIVES: Considering that the examination of psychological flexibility measures among older adults is scant, this study explores the psychometric properties of the Acceptance and Action Questionnaire-II (AAQ-II) among outpatient geriatrics primary care clinic patients in a university-affiliated medical center in the Deep South. METHODS: On average, patients who consented to participate in research (N = 119) were 75.42 (SD = 9.14) years old. Approximately 74% were women, 91% were non-Hispanic white, and 86% lived in urban areas. Measures included psychological inflexibility, subjective health literacy, depression, anxiety, and global cognitive functioning. RESULTS: The AAQ-II demonstrated adequate internal reliability, Cronbach's α = 0.79, and good test-retest reliability, r(28) = .84, p < .001. With the removal of item 4, confirmatory factor analysis demonstrated adequate construct validity for the use of AAQ-II with older adults. As predicted, the AAQ-II exhibited concurrent, predictive, discriminant, and incremental validity within this older adult sample. CONCLUSIONS: These results demonstrate meaningful scientific and clinical implications. This is the first study to examine the psychometric properties of the Acceptance and Action Questionnaire-II (AAQ-II) in an older adult sample. CLINICAL IMPLICATIONS: The AAQ-II is a psychometrically sound measurement that can be clinically used on older adults.

Centering racial justice for Black/African American and Indigenous American people in commercial tobacco product regulation.

Although overall health in the United States (US) has improved dramatically during the past century, long-standing health inequities, particularly the unequal and unjust burden of tobacco-related disease and death among racialized populations, persist. A considerable gap exists in our understanding of how commercial tobacco product regulations and policies cause and/or exacerbate race-based health inequities among Black/African American (B/AA) and Indigenous American people. The purpose of this paper is to 1) describe how existing US commercial tobacco regulatory policies may contribute to structural racism and undermine the full benefits of tobacco prevention and control efforts among B/AA and Indigenous American groups; and 2) initiate a call to action for researchers and regulators of tobacco products to examine policies using an equity lens. These actions are imperative if empirically-informed regulation of commercial tobacco products is to address health equity.

Educating the Public on the Health Risks of Very Low Nicotine Content Cigarettes: Results From a US-Based Convenience Sample.

INTRODUCTION: US FDA issued an advance notice of proposed rulemaking to reduce nicotine in cigarettes. To maximize the benefits of this potential standard, very low nicotine content (VLNC) cigarettes must be communicated in a way that does not result in misperceptions. AIMS AND METHODS: Adults (n = 567 who smoke; n = 610 non-smokers) from an online platform were randomized to a control message previously associated with accurate addictiveness perceptions of VLNC cigarettes but health misperceptions or to one of five messages that also included messaging on nicotine morbidity effects or VLNC cigarettes morbidity or mortality effects. p value <.01 was significant. RESULTS: In participants who smoke, perceived lung cancer risk (responses: 1, very little risk to 10, very high risk) if smoked VLNC cigarettes regularly was higher in conditions that communicated mortality effects of VLNC cigarettes compared to the control (7.12-7.18 vs. 5.97, p values < .01). In non-smokers, perceived lung cancer risk was higher in all five message conditions when compared with the control (7.58-8.22 vs. 6.35, p values < .01). Proportion who responded accurately (ie, False) to the statement Cigarettes with 95% less nicotine are safer than cigarettes with normal nicotine levels was higher in conditions describing VLNC morbidity or mortality effects when compared with the control in both participants who smoke (52.04-67.37% vs. 30.85%, p values < .01) and do not smoke (62.50-72.38% vs. 32.00%, p values < .01). CONCLUSIONS: Messaging on mortality effects of VLNC cigarettes (ie, cigarettes with 95% less nicotine are as deadly as current cigarettes) was associated with more accurate perceptions of the health risks of VLNC cigarettes than the control; however, misperceptions remained in one-third of participants. IMPLICATIONS: One approach to communicating a VLNC cigarette standard to the public is to include messaging on the mortality effects of VLNC cigarettes. However, further study and possible refinement of this message condition are recommended since approximately one-third of participants exposed to this message still perceived VLNC cigarettes to be safer than normal nicotine content cigarettes.

Racial/ethnic group comparisons of quit ratios and prevalences of cessation-related factors among adults who smoke with a quit attempt.

BACKGROUND: Smoking-related disparities exist among racial/ethnic minoritized groups. OBJECTIVE: We compared quit ratios and smoking cessation-related protective and risk factors by race/ethnicity to inform approaches to reduce disparities. METHODS: Among adults who smoke with a quit attempt from Wave 4 (2016-2017) Population Assessment of Tobacco Use and Health Study, the following factors were examined by racial/ethnic group (American Indians/Alaska Native [AI/AN;n = 165], Black/African American [AA;n = 526], Asian [n = 38], Hispanic/Latino/Latina/Spanish [n = 475], or White [n = 1,960]), wherein each were nearly gender-balanced: cessation medications, counseling/self-help materials, home smoking ban, social support, e-cigarette use, sleep, and mental health. RESULTS: Quit ratio was lower for AI/AN (adjusted odds ratio[aOR]:0.61) and Black/AA (aOR:0.49) and higher for Asian (aOR:1.90) and Hispanic/Latino/Latina/Spanish (aOR:1.30) than White adults. Medication use was low among all and lower among Black/AA (aOR:0.70) and Hispanic/Latino/Latina/Spanish (aOR:0.56) than White adults. Use of counseling/self-help materials were low among all and higher in AI/AN (aOR:1.85), Black/AA (aOR:1.87), and Hispanic/Latino/Latina/Spanish (aOR:1.49) than White adults. Presence of a smoking ban was lower among Black/AA (aOR:0.40) and higher in Hispanic/Latino/Latina/Spanish (aOR:1.59) than White adults. E-cigarette use was lower in Black/AA (aOR:0.53) and Hispanic/Latino/Latina/Spanish (aOR:0.43) than White adults. Sadness, anxiety, and sleep difficulties were higher in AI/AN (aORs:1.57, 1.50, 1.64) than White adults. CONCLUSIONS: All racial/ethnic groups would benefit from policies and programs that increase cessation medications and counseling. Quit ratios were particularly low among Black/AA and AI/AN adults. Black/AA adults may benefit from efforts to increase smoking bans, while AI/AN adults may benefit from cessation approaches that simultaneously target sleep and mental health.

Nucleosomes inhibit target cleavage by CRISPR-Cas9 in vivo.

Genome editing with CRISPR-Cas nucleases has been applied successfully to a wide range of cells and organisms. There is, however, considerable variation in the efficiency of cleavage and outcomes at different genomic targets, even within the same cell type. Some of this variability is likely due to the inherent quality of the interaction between the guide RNA and the target sequence, but some may also reflect the relative accessibility of the target. We investigated the influence of chromatin structure, particularly the presence or absence of nucleosomes, on cleavage by the Streptococcus pyogenes Cas9 protein. At multiple target sequences in two promoters in the yeast genome, we find that Cas9 cleavage is strongly inhibited when the DNA target is within a nucleosome. This inhibition is relieved when nucleosomes are depleted. Remarkably, the same is not true of zinc-finger nucleases (ZFNs), which cleave equally well at nucleosome-occupied and nucleosome-depleted sites. These results have implications for the choice of specific targets for genome editing, both in research and in clinical and other practical applications.

Tobacco harm reduction: Past history, current controversies and a proposed approach for the future.

Tobacco harm reduction remains a controversial topic in tobacco control. Tobacco harm reduction involves providing tobacco users who are unwilling or unable to quit using nicotine products with less harmful nicotine-containing products for continued use. The skepticism towards harm reduction is based in part on the experience with low-yield tar/nicotine cigarettes, which were presumed to be associated with lower health risks than higher yield cigarettes and marketed as such by cigarette manufacturers. Only later did the field learn that these cigarettes were a deceptive way for cigarette manufacturers to allay the health concerns over cigarette smoking. Since this experience, there has been a proliferation of tobacco products that might potentially serve as a means to reduce tobacco harm. Some members of the tobacco control community believe that these products have great potential to reduce mortality and morbidity among smokers who completely switch to them. Others believe that we will be addicting another generation to tobacco products. This paper reviews the past history, the current tobacco landscape and controversies, and an approach that might rapidly reduce the yearly half-million deaths associated with cigarette smoking in the U.S.

Relationships Between Race, Gender, and Spot Urine Levels of Biomarkers of Tobacco Exposure Vary Based on How Creatinine Is Handled in Analyses.

INTRODUCTION: We illustrate the differential impact of common analysis approaches to handling urinary creatinine, a measure for urine dilution, on relationships between race, gender, and biomarkers of exposure measured in spot urine. METHODS: In smokers, spot urine levels of total nicotine equivalents (TNE, sum of total nicotine, total cotinine, and total 3'-hydroxycotinine) and total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) overall and per cigarette were examined. Relationships between race (African Americans [AA] n = 373, Whites n = 758) or gender (males n = 629, females n = 502) and TNE or NNAL were examined using the following approaches to handling creatinine: (1) unadjusted/unstandardized; (2) standardization; (3) adjustment as a covariate. Significance was considered at p < .05. RESULTS: Creatinine was higher in AA versus Whites (1.19 vs. 0.96 mg/mL; p < .0001) and in males versus females (1.21 vs. 0.84 mg/mL; p < .0001). Independent of how creatinine was handled, TNE was lower among AA than Whites (TNE ratios AA vs. Whites: 0.67-0.84; p's < .05). Unadjusted TNE per cigarette was higher among AA versus Whites (ratio 1.12; p = .0411); however, the relationship flipped with standardization (ratio 0.90; p = .0360) and adjustment (ratio 0.95; p = .3165). Regarding gender, unadjusted TNE was higher among males versus females (ratio 1.13; p = .0063), but the relationship flipped with standardization (ratio 0.79; p < .0001) or adjustment (ratio 0.89; p = .0018). Unadjusted TNE per cigarette did not differ across gender (ratio 0.98; p = .6591), but lower levels were found in males versus females with standardization (ratio 0.68; p < .0001) and adjustment (ratio 0.74; p < .0001). NNAL displayed similar patterns. CONCLUSIONS: Relationships between race, gender, and spot urine levels of biomarkers of exposure can vary greatly based on how creatinine is handled in analyses. IMPLICATIONS: Lack of appropriate methods can lead to discrepancies across reports on variability of urinary biomarkers by race and gender. We recommend that for any analyses of biomarkers of exposure measure in spot urine samples across race, gender, or other population subgroups that differ in urinary creatinine levels, sensitivity analyses comparing the different methods for handling urinary creatinine should be conducted. If methods result in discrepant findings, this should be clearly noted and discussed.

Development and Piloting Testing of an Experimental Tobacco and Nicotine Product Marketplace.

INTRODUCTION: We describe the development and pilot testing of the experimental tobacco and nicotine product marketplace (ETM)-a method for studying tobacco and nicotine product (TNP) choices and use behavior in a standardized way. AIMS AND METHODS: The ETM resembles an online store populated with TNPs. Surveillance activities and data from a US representative survey and consumer reports were used to determine the most popular TNPs for inclusion in the ETM. Standardized information and videos demonstrating how to use the TNPs were provided. To test the feasibility of using the ETM, smokers (n = 119) underwent monitoring of usual brand cigarette smoking and other TNP use (Baseline Phase) followed by access to the ETM (ETM Phase) that included their usual brand cigarettes, e-cigarettes, moist snuff, snus, and nicotine replacement therapy. During the ETM Phase, participants were provided points based on their baseline TNP consumption to exchange for TNPs in the ETM. Participants were advised to exchange points for enough TNPs to last until their next visit and to refrain from using TNPs not obtained in the ETM. A subset of the participants (n = 62) completed a survey on their experience with the ETM. RESULTS: The majority of the participants stated they were comfortable with navigating the ETM (97%), it was easy to determine product characteristics (89%), and they were satisfied with the products included in the marketplace (85%). CONCLUSIONS: The ETM was well received by the vast majority of the participants and can be utilized by researchers to investigate a variety of TNP policy and regulatory science research questions. IMPLICATIONS: Patterns of TNP use are complex due to greater availability, marketing, and promotion of a diverse array of TNPs. Innovative methods are needed to experimentally study TNP choices and patterns. Through describing the development of the ETM, we provide researchers with a tool that can be readily adapted to studying a variety of phenomena challenging public health.